HISTOLOGY LECTURE NOTES

GENERAL HISTOLOGY

1. INTRODUCTION

Histology is a branch of science concerned with the microscopic structure of cells,

tissues, and organs in relation to their function..

Synonyms: microanatomy, microscopic anatomy, cell and tissue biology

Tissues are groups of cells and their associated extra-cellular substances which are
specialized to carry out specific functions [Fr. Tissue = fabric, texture, weave].

Cell is the smallest structural and functional unit of any living organsim. Unique
features of any cell includes its ability to divide and to produce energy for its survival. Most of
the living creatures are unicellular (i.e., madeup of a single cell), whereas, the multicellular
organisms are composed of different types of cells that are evolved to perform dif ferent
functions of that organism viz., secretion, absorption, contraction, conduction and excretion.
Structure of any eukaryotic cell includes two major compartments viz., cytoplasm
and nucleus. The cell is delimited by the cell membrane. Cytoplasm consists of organelles,
inclusions, and cytoskeletal components enclosed in a semiviscous liquid, the cytosol. The
study of the structural components of the cell is referred to as Cytology, while the study of
the integration of cells to form tissues and organs is referred to as Histology [Gr. histos =
web, tissue ; logos =study of]. Such integration involves three different components: the
cells, the extracellular matrix (ECM), and tissue fluids. The ECM components are
synthesized by cells and are present in different proportions in different tissues. Tissue fluids
transport nutrients, hormones, gases, and waste products to and from the cells.

METHODS OF STUDY

Microscopic examination:
In the higher mammals, cells have a dimension of few hundredth of a millimeter. The
Greek letter "µ" is used as a symbol. Hence, many parts of the cell can be studied with the
help of light microscope, which allows maximal resolution of structures down to 0.2 µm in
diameter at magnifications up to 1000 to 1500 times. However, many cellular structures are
smaller and the electron microscope, offering much better resolution (down to 0.1 nm) and
higher magnifications (up to X400,000), has to be used for their study. This includes both
transmission electron microscope (TEM) and the scanning electron microscope (SEM).
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Now the atomic force microscope (AFM) can also provide high-resolution images, which
are comparable in resolution to those obtained from TEM. Both EM and AFM, because of
their greater resolution and useful magnification, are often the last step in acquiring high
resolution details of cell structure.
STAINS AND STAINING
In order to visualize the celllular details, it is necessary to stain cells and tissues to
increase the contrast under light or electron microscope. Staining is the process by which
the cell structures are coloured differently by using chemicals called stain. A stain or dye is
a chemical substance used to impart colour to the cell structure or tissues to facilitate their
examination and identification.There are two types of stain. 1. Acid Stain:
A stain in which the coloring agent is in the acid radical.It stain the basic components of cells
Eg. Eosin.2. Basic Stain: A stain in which the coloring agent is in the basic radical.It stains
the acidic components of cells. Eg. Hematoxylin.
Acidophilia

It refers to the capacity of cell structures to be stained by acid stain viz., eosin, acid
fuchsine, picric acid and others. Such structures are called acidophilic or eosinophilic. The
cause of acidophilia is principally due to the basic properties of the cell structure.

Basophilia

It refers to the ability of the cell structures to be stained by basic dyes viz.,
haematoxylin. Structures stained by basic stain is known as basophilic. Basophilia caused by
acidic nature of the cellular component. In the cell, cytoplasm is generally basic and nucleus
is generally acidic in nature.

Most commonly used stains in staining histological sections include Haematoxylin

and Eosin (H&E). Of which, Haematoxylin is a basic dye which stains the nucleus with blue
colour. Eosin is an acidic stain which stains the cytoplasm with pink colour.

Vital staining. Vital stains are employed to color certain components within living
organisms without doing harm to the vital tissues. For example, trypan blue and India ink
form colloidal suspensions in water. Since these large particulates will not diffuse into cells,
they will be cleared from the body by the phagocytic activity of macrophages, and are thus
excellent markers for members of the mononuclear phagocyte system (MPS). Supravital
staining is a variant of this procedure in which these dyes are applied to living tissues that
have first been removed from the organism by, for example, biopsy.
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A number of other special methods are used in this study of tissues like histochemical
methods, radio-autography, fluorescence microscopy, micro incineration, micro dissection
and tissue culture etc.

Metachromasia

Certain basic dyes react with tissue components that shift their normal color from blue
to red or purple; this absorbance change is called metachromasia.The underlying
mechanism for metachromasia is the presence of polyanions within the tissue. When these
tissues are stained with a concentrated basic dye solution, such as toluidine blue , the dye
molecules are close enough to form dimeric and polymeric aggregates. The absorption
properties of these aggregations differ from those of the individual nonaggregated dye
molecules. Cell and tissue structures that have high concentrations of ionized sulfate and
phosphate groups—such as the ground substance of cartilage, heparin-containing granules
of mast cells, and rough endoplasmic reticulum of plasma cells—exhibit metachromasia.
Therefore, toluidine blue will appear purple to red when it stains these comp onents

Microscopy

Cells are the smallest units of living organisms. Normally, a typical animal cell is
about 10-20 micrometre in diameter and therefore can not be visualized by naked eye.
Hence, light microscope is used to examine sections under high magnifications.
The usefulness of any microscope is dependent not merely upon its ability to magnify but
upon its ability to resolve. The maximum useful magnification of an ordinary light microscope
is 1500 time.

The Electron Microscopy (EM) that uses electron beams instead of light and electro
magnetic fields in place of lenses is used to study tissues under very high magnification of
5,000 to 20,000. The image is usually recorded on a photographic plate and further increase
in magnification is achieved by enlargement. This technique involves costly and complicated
equipment and sectioning has to be done under vacuum, which may produce considerable
distortion, and the sections should be 0.1 micron thick or even less.

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2. THE CELL

Histology is primarily the study of the arrangement of differentiated cells into

tissues. Cell biology comprises essential background knowledge for studying histology. It is
presumed that the familiarity with basic cell structure and function will help in bette r
understanding the organization of tissues and organs of the animal body.

All living things are composed of cells and cell products. This insight is one of
the greatest unifying principles of biology. Each of the body's cells contains all of the parts
and processes necessary for life and is potentially an independent organism. (Some, like
macrophages, are more independent than others!) All the bodily functions of any multicelluar
organism is made possible from the activities and interactions of these individual cells.

All diseases result from disorders in cellular function. This hypothesis was put
forward by Rudolf Virchow, the father of modern pathology, in the middle of the
nineteenth century, soon after the establishment of the Cell Doctrine.

"Life itself is but the expression of a sum of phenomena, each of which follows the
ordinary physical and chemical laws. Disease is not something personal and special, but only
a manifestation of life under modified conditions, operating according to the same laws as
apply to the living body at all times, from the first moment until death." ( Rudolf Virchow).

CELL STRUCTURE

The cell consists of two compartments namely the cytoplasm and nucleus.
Cytoplasm refers to the protoplasm of the cell body and Karyoplasm denotes the
protoplasm of the nucleus. The protoplasm contains high proportion of water with proteins,
carbohydrates, fat and inorganic salts. It is in the form of colloidal mass dispersed in water
and may vary from a state of hydrogel to hydrosol either in different type of cell or in the cell
under different physiological conditions. The karyoplasm is more of gel the cytoplasm is more
of sol in character.

The proteins of biological importance are (1) Simple proteins: Albumins, globulins
(2) Conjugated proteins: Nucleo-proteins, glycoprotein or mucoprotein, lipoproteins, chromo
proteins. The various enzymes found in and produced by the cells are all made up of
proteins.

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Proteins of Nuclei are of great importance in the study of all aspects of cell activity.
They are combinations of nucleic acids with proteins. There are two kinds of nucleic acids,
one is DNA or Deoxyribonucleic acid and the other is RNA or Ribonucleic acid. DNA is found
in the nucleus more specifically, in the chromosomes and thus it is important in heredity.
RNA is found both in the nucleus and the cytoplasm, particularly in the latter. It is abundant
in cells showing high metabolic activity.

The Carbohydrates of significance are glucose and glycogen and also neutral and
acid mucopolysaccharides which are present in the intercellular substance.

Most of the Fat in the body is in the form of droplets of neutral fat stored in special fat
cell. More complex lipids such as sterols, phospho-lipids are present in certain cell.

The inorganic constituents of protoplasm are chiefly potassium; small amounts of

magnesium little sodium and less calcium. Of the anions bicarbonates and phosphates
predominate. In the intercellular fluid sodium and chlorides predominate. Concentrations of
one element may be found in certain tissues like calcium in bone, iodine in thyroid gland and
iron in erythrocytes etc.

STRUCTURE OF THE CELL

The cells in the body vary greatly in size, shape and structure and these various
adaptations are for the different functions which the cells perform in various tissues of the
organs. However most of the cells retain a number of features in common and these general
features are now described.

Plasma membrane or cell membrane: This covers the surface of the cell and consists
of a semi permeable membrane, through which the interchange of materials between the cell
and its environment occurs. It is composed of a continuous layer of lipid molecules in
between two protein layers. These layers could be distinguished under electron microscope
and under ordinary light microscope; the plasma membrane is too thin to be distinctly seen.

Cytoplasm

Cytoplasm contains a number of formed bodies embedded in a substance which

appears translucent and homogenous in the living cell. This substance called the ground
substance of the cytoplasm assumes various appearances after precipitation or coagulation

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with fixatives. It may appear reticular, fibrillar or granular depending upon the cell and the
technique used.

Formed bodies in cytoplasm: These are two groups (1) Organoids or organelles
composed of differentiated cytoplasm which performs some special function related to
metabolic or other activities of the cell. (2) Inclusions which are metabolic products or
ingested substances not playing direct part in the metabolic activity of the cell.

Organelles: These include (1) Centrosphere and centriole (2) Mitochondr ia

(3) Ribosome, (4) Endoplasmic reticulum and chromophil substance, (5) Golgi
apparatus, (6) Lysosomes and (7) Fibrils.

1. Centrosphere and centriole (Central body, centre): This lies close to the nucleus
and is present in all mammalian cells that can divide, but is difficult to demonstrate in certain
types of cells. It consists of one or two sharply staining granules the centre surrounded by a
spherical area of dense cytoplasm called the centrosphere or centrosome. The centrosphere
and centrioles play an important part in the early stage of mitosis (cell division) (see details
under mitosis). They are also associated with basal granules of motile cilia and flagella.

2. Mitochondira: These appear as minute granules, rod or filaments under light

microscope and are demonstrable by special methods. Electron microscopic studies show
that the mitochondria are bounded by double membranes, viz., an outer and an inner and the
substance or matrix within is partially subdivided by a series of parallel ridges or cristae,
which protrude inwards from the inner memberane. Mitochondria contain numerous
enzymes (Respiratory enzymes esp. those of kreb’s citric acid cycle, enzymes catalyzing
Oxidative phosphorylation etc) and hence mitochondria play an important role in cell
respiration and other Oxidative processes in the cell by which energy is released for other
metabolic processes in the cell.

3. Ribosomes (Cytoplasmic RNA): These appear under electron microscope as fine

cytoplasmic granules or particles and are composed of RNA (Ribonucleic acid) and
associated proteins. The Ribosomes are present in most of the cells (excepting mature
erythrocytes) and their presence gives rise to basophilia of the cytoplasm visible under light
microscope. The Ribosomes or RNA granules may be diffusely distributed in the cytoplasm
or they may be attached along the outer surfaces of the membranes of Endoplasmic
reticulum. The basophilia may be diffuse when the R.N.A. granules are uniformly distributed
in the cytoplasm (devoid of any association with endoplasmic reticulum) or may be localized
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or patchy when they are associated with endoplasmic reticulum. The ribosomes play an
active role in the synthesis of proteins for secretion for renewal and maintenance of
cytoplasmic protein and are used for meeting the requirements of rapidly growing and
dividing cells.

4. Endoplasmic reticulum and chromophilic substance: This is visible only under

electron microscope and consists of a network of double membrane lined spaces, which form
interconnected network and tubules extending throughout the cytoplasm. Two types are
recognized. (a) Rough surfaced and (b)Smooth surfaced.

In the rough surfaced variety, numerous ribosomes are studded along the outer
surface of membranous vesicles and the rough surfaced vesicles are to segregate protein
secretions, enzymes etc. within the membrane bound cisterns following the synthesis at the
sites of ribosomes on the outer surface of the membrane.

Under light microscope, the areas of the cell occupied by rough surfaced vesicles
show pronounced localized or patchy basophilia and it was attributed to the presence of
chromophilic substance in those cells (Nissl bodies in nerve cells). It was also referred to as
Ergastoplasm in acinar cells of pancreas. It has now been established by electron
microscopic studies that the chromophilic substance and ergastoplasm are all due to the
presence of rough surfaced vesicles of endoplasmic reticulum and the basophilia is due to
the RNA granules on outer surfaces of the membranes as the membranes of the reticulum
are not basophilic.

Smooth surfaced endoplasmic reticulum is found in many different types of cells, like
skeletal muscle fibres, in cells synthesizing steroid hormones and in cells engaged in
detoxification. It consists of a network of membranous and tubular structures but devoid of
Ribosomes on their outer surface.

5. Golgi apparatus (Internal reticular apparatus of Golgi): It was first described by

Golgi, as a dark network in Nerve cells by silver impregnation technique. The network was
subsequently demonstrated in many cells by similar special technique and it is usually
located in the same pole as the cell centre or in secretory epithelial cells between the
nucleus and free border. It is composed of lipoproteins. Under electron microscope, it
appears as a collection of double membranes forming vesicles which appear to be of three
types 1.Flattened vesicles 2. Secretory vesicles and 3. Micro vesicles. The flattened vesicles

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are elongated closely packed together. The peripheral portions of these flattened vesicles
may become distended and then bud off to form secretory vesicles. The secretory vesicle
granules are discharged from the cell as the secretion. The micro -vesicles are small
membranous vesicles and are found in the region between the endoplasmic reticulum and
the flattened vesicles. It is believed that the protein material is synthesized in the rough
surfaced vesicles from where they are budded off as secretory vesicles.

The Golgi apparatus does not in itself produce the entire secretion but serves to
concentrate the secretion and to package them before elimination from the cells. There is
some evidence that the carbohydrate components of some secretions may be produced by
the Golgi apparatus.

6. Lysosomes: These are small membranous vesicles containing lytic enzymes and
are concerned with lysis or dissolution of cells, with lysis of phagocytosed material which
cannot enter into direct chemical reactions in the cytoplasm.

7. Fibrilla: Definite fibrillae are characteristic and prominent structures in muscle and
nerve cells and are concerned with the specialized functional activity of such cells.

8. Inclusions: These are stored products in the cytosol which results from metabolic
activity of the cells. These may be fats, yolk and glycogen. Pigments may be exogenous or
endogenous.

9. Ctoskeleton: The term cytoskeleton is misleading in that it covers structures that

function to maintain not only cell shape, but also the motility and intracellular transport
functions of the cell. There are three main components of the cytoskeleton: microtubules,
microfilaments, and intermediate filaments.

Cytosol:

The cytosol makes up about half of the cell volume and contains water, ions, sugars,
amino acids, nucleotides, hundreds of soluble enzymes (e.g., enzymes of glycolytic
pathway), cytoskeletal components, messenger RNA, transfer RNA, and group of other
molecules. Polymerization of actin is involved in regulating the viscosity of the cytosol.
Transition of the cytosol from a gel state to a more soluble (gel-to-sol) state assists the
formation of extensions (pseudopodia) involved in cell motility.

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NUCLEUS

The nucleus is contained within a thin but definite nuclear membrane more distinct
than plasma membrane under electron microscope. The nuclear membrane is seen to
consist of two layers and in certain places openings or nuclear pores appear, which may
facilitate exchange between nucleus and cytoplasm.

The nucleus contains nucluear sap (karyolymph) and in it are the nucleolus and
chromatin. The structural appearance of the nucleus undergoe s considerable alteration
during mitosis or cell division.

The nucleolus is generally rounded in shape and is a dense well defined body,
although no limiting membrane is present. They are composed of RNA and associated
protein and may exhibit basophilia or acidophilia in different cells depending on the
proportions of RNA and basic protein. Under electron microscope, many fine granules are
within the nucleoli which are generally arranged in irregular rows or networks sometimes
described as Nucleolonema. The nucleolus is associated with particular region of a particular
chromosome. It disappears during prophase and reappears during telophase of mitosis. The
nucleolar material is apparently derived from all chromosomes but accumulates in
association with particular chromosomes in a region known as the nucleolus organizer.

The chromatin consists of clumps, or granules or threads which stain with basic dyes.
They are composed of DNA and associated proteins and they represent parts of
chromosomes.

Chromosomes of the nucleus appear as distinct thread-like structures during cell

division and take an intense basic stain. When a cell begins to divide the structural
components of each chromosome became contracted and tightly coiled. During the resting
stage or interphase the chromosomes become elongated and loosely twisted interspersed
with tightly coiled dense regions, in ordinary light microscope, the elongated portions are not
distinct but the dense, tightly coiled portions appear as granular masses, referred to a s
chromatin.

A fairly distinct body named sex-chromatin can be seen in the cells of females. It is
usually seen as a small Plano convex spot on the inner side of the nuclear membrane. It is
due to the presence of two ‘X’ chromosomes in the female.

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Fig.2.1 Interphase nucleus

1. Nuclear envelope
2. Nuclear pore
3. Nucleolus
4. Endoplasmic reticulum
5. Heterochromatin
6. Euchromatin

Chromosomes become distinct during cell division. They contain DNA and associated
protein and a small quantity of RNA. They carry genes which are the actual determiners of
hereditary characters. The genes lie in definite organization in the chromosomes of
hereditary characters. The genes lie in definite organization in the chromosomes and have
their chemical basis in DNA. During mitosis there is duplication of the chromosomes and the
total DNA content of the cell. Mature germ cells-contain only half the number of
chromosomes, having undergone meiosis or reduction division, in all other cases the number
of chromosomes and consequently DNA content in each cell is constant for a particular
species. The number of chromosomes in different species is as follows; man - 46; ox-60;
horse-60; sheep-60; dog-78; cat-62; pig-40.

Functions of the nucleus: a) It is important for cell division b) The presence of

chromosomes in the nucleus is responsible for transmission of hereditary characters.
c) It controls and regulates all metabolic activities carried on in the cytoplasm.
d) The nucleus and nucleoli play an important role in synthesis of cytoplasmic proteins.
e) The nucleus and cytoplasm are interdependent. The cell dies when the nucleus is
removed and an isolated nucleus is not viable.

Cell size: Varies from 3 to 20 microns in general. The largest cells are the ova which
reach a size of 100 to 200 microns. The smallest cells are lymphocytes measuring
3 to 5 microns. The shape and form of the cells in different tissue vary greatly. This may be
circular, discoid, oval, squamous, cuboidal, columnar, spindle shaped , pyramidal or stellate
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shaped etc. The shape of the nuclei also vary correspondingly and may be spherical, oval,
elongated, rod shaped, flattened or polymorphous showing many lobes. Some cells may
have two or more nuclei. Based on the shape and type of cell, location of nucleus varies.

Functions of the Protoplasm:

Protoplasm shows the following vital properties:

(a) Metabolic activities, essential for the life of cell,

(b) Special functions (eg. irritability such as conduction, contraction, etc.)

(c) Independent movement (amoeboid motion, ciliary or flagellar movement) and

(d) Growth and reproduction.

CELL DIVISION

Cell division is the only source of living cells. There is no spontaneous generation.
Cell division, the most important activity of cell life, is accomplished by an equal division of
the nucleus and cytoplasm, called mitosis.

MITOSIS

The division of the cytoplasm immediately follows that off nucleus. Mitosis,
(karyokinesis or indirect cell division) exhibits a regular sequence of events, which come
under four phases-prophase, metaphase, anaphase and telophase.

Prophase: Cells about to divide assume a more spherical shape. The nucleus
enlarges and long slender intertwined double filaments become visible. These are the
chromosomes. Chromosomes become progressively shorter and thicker. The nuclear
membrane disappears. The nucleoli also disappear. The centrioles move into the vicinity of
the nucleus, if centriole is single it divides into two. The lighter area surrounding centrosome,
called centrosphere shows radiations which spread out to the periphery and is called aster.
The centrioles move apart surrounded by their own aster. Now a bundle of fibrils appears
between them. These fibrils become long and form a central spindle. The two centrioles
move to the opposite poles. The spindle moves into the centre of the cell and the
chromosomes occupy the equatorial plane of the spindle.

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Metaphase: This is a short period when all the chromosomes are arranged around
the spindle in the equatorial plane. Each chromosome is split longitudinally into two exact
halves called chromatids. Chromosomes now are typically V, I, J or U shaped. The free ends
of the chromosomes stick out to the edge of the equatorial plane.

Anaphase: This is the process of equal distribution of the halves of the chromosomes
the spindle persists. As the chromosomal groups reach the centrioles, the srage is set for
organization of daughter nuclei. The chromosomal groups are identical in all respects.

Telophase: In this phase the chromosomes aggregate more closely; their outline
gradually becomes blurred. The cell body is divided by an annular furrow which deepens,
until it, constricts the cytoplasm into two halves at the level of equatorial plane. The nucleoli
are formed, as also the nuclear membrane and typical chromatin pattern of interphase nuclei.
The spindle disappears and centriole is now doubled. The duration of mitosis is half to one
hour.

Amitosis or direct division of nucleus without appearance of chromosomes is rare in

higher animals.

Meiosis or reduction division is a special kind of division observed in germ cell

resulting in the reduction of chromosomes to the haploid number of the species.

CELL CYCLE

Somatic cells multiply by mitosis and as a result two identical daughter cells are
produced. Any somatic cell, before it undergoes mitosis, it must duplicate its content of
genetic information so that the resulting two daughter cells will contain the same amount of
DNA as the parental cell. In addition, since the cytoplasm and organelles are also divided
between the daughter cells during cytoplasmic division, it is also necessary for the parental
cell to increase in size through growth before division. The cell therefore undergoes a series
of definable steps referred to as the cell cycle. These steps includes G1 phase (gap1), S
phase(synthesis of DNA), G2 phase (gap2), and M phase (mitosis). During G1 phase, the
cell grows and receives signals (growth factors) from its surroundings which order the cell to
commence preparations for mitosis. While in the S phase, doubling of its DNA takesplace.
During G2 phase, the cell grows further, checks the quality of the DNA it replicated during

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the S phase, and synthesizes proteins (such as tubulin subunits) needed for the M phase.
After mitosis, the daughter cells may enter either a new cell cycle or a resting stage (G 0).

Fig.2.2 Stages of cell cycle

Ordered transition through each of these steps is required. At critical checkpoints, the
cell will determine if it is of the correct size and if DNA has been correctly replicated. Before
proceeding to the next step of the cell cycle, appropriate signals must be received from
neighboring cells. Progression is controlled by special proteins called cyclins. Different
cyclins are needed at different checkpoints and will activate enzymes (cyclin-dependent
kinases, or CDKs) that make it possible for a cell to continue to the next step. Thus, the cell
cycle is tightly controlled. Failure of the cell to produce the right cyclin at the right time will
stop cell cycle progression and may even result in the death of the cell through apoptosis. If
the cell cycle progresses, the cell will enter mitosis.

☺☺☺☺☺

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3. TISSUES
A tissue is an association of cells and intercellular substance specialised to perform
specific function. Intercellular substance or extracellular matrix (ECM) is in between the cells
and produced by the cells themselves. The process of by which the tissue is able perform
specific functions is called cellular differentiation.

ECM or intercellular substance generally consists of a homogenous ground

substance (also referred as intercellular cement substance) and in certain tissues, fibres may
also be found, embedded in the ground substance.

The structure, shape, size, and arrangement of the cells, the bio -chemical and
physicochemical properties of ECM, and the nature of fibres when present are different in the
various types of tissues. Further the proportion between the cells and the intercellular
substance also differs from one type of tissue to another.

There are four or sometimes stated as five types of elementary or fundamental

tissues in the adult animal body. Organ systems are made up of combinations of these
fundamental tissues arranged in a specific and characteristic way in each organ.

The elementary or fundamental tissues of the body are,

1. Epithelium

2. Connective tissue

3. Muscular tissue

4. Nervous tissue and

5. Sometimes blood and lymph are classified together as a separate fundamental

tissue.

The same 4 basic tissue types are found in all multicellular animals: epith elium,
connective tissue, muscular and nervous tissue.

The basic subtypes of all fundamental tissue are found throughout in most
vertebrates, and are combined in the same way to form organs.

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EPITHELIUM

Epithelium is one of the fundamental tissue in the body composed primarily of cells,
between which is a minute amount of intercellular material. Closely approximated cells held
together by a small amount of viscous substance known as intercellular cement or matrix.
Epithelial tissue always presents a free surface. Epithelium is avascular and receives its
nutrients by difusion of molecules through the basement membrane with which it is attached.

Epithelium originates from all the three germ layers i.e ectoderm, endoderm and
mesoderm. Epithelium covers the body surface and lines all tubular passages leading to the
exterior, lines various body cavities and constitutes the secretory epithelium in all exocrine
glands. Between the epithelium and the underlying connective tissue, there is usually exist a
basement membrane, a condensation of the intercellular substance to the connective tissue.

Shape and arrangement of cell: The shape of epithelial cells varies from a very flat
type (squamous) to tall rod-like columnar cell and intermediate between these two are
cuboidal, which may be low or high. When an epithelium is lined by one layer of cells, it is
called simple epithelium; when it is lined by two are more layers of cells it is defined as
stratified epithelium. In the stratified epithelium, the basal layer of cells rest upon the
basement membrane and cells in the topmost layer have a free surface. In certain places,
the nuclei of epithelial cells lies at various levels giving a stratified appearance. But in these
cases, all cells reach the basement membrane though all do not have a free surface. This
type of epithelium is called pseudostratified epithelium and is characteristic of respiratory
passages and reproductive passages.

Attachment between cells: The cells of epithelium are attached to each other by
adhesions of the cell membranes of adjacent or opposing cells. At these points of adhesions,
the membrane of each cell is thickened and under EM, fine fibrils are seen in the adjacent
areas of cytoplasm but there is no protoplasmic continuity between the cells. These
thickened areas of adhesion are known as desmosomes with light microscope.
Adhesions in stratified epithelium are named as intercellular bridges. This is due to
shrinkage caused by fixatives.

Modification of cell surface : The cytoplasm of cells at the free surface may show
modifications. The cells of intestinal epithelium show a pronounced modification of free
surface. They have a definite banded layer of uniform thickness known as the striated border

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in which striations are vertical to the surface. These striations have been found to be
microvilli under EM. This adaptation increases the surface area available for absorption.
The cells of kidney tubules (proximal convoluted part) show brush border to facilitate
selective reabsorption.

The apical surface of epithelial cells lining respiratory passages and epithelium lining
Fallopian tubes shows motile cilia - kino cilia. The ciliary movement occurs in regular waves.
The movement of the cilia consists of a rapid effective beat and a slow recovery stroke,
always in one direction (usually towards the external opening of the tube or passage).
Whereas, parts of male reproductive tract have epithelial cell with non -motile cilia - stereo
cilia. These are supposed to help in elimination of secretion from the cells.

Basement membrane (Membrana propria): This is a sheet of variable thickness,

interposed between the epithelium and subjacent connective tissue. It serves for attachment
of epithelium to the underlying connective tissue. It consists of a ground substance of the
underlying connective tissue. Embedded in this condensed ground substance are delicate
reticular fibres, which are continuous with those of underlying connective tissue . Basement
membrane in ordinary preparations are distinguishable only in some locations (eg.trachea).
In other locations, beneath the stratified squamous epithelium and columnar epithelium of the
intestine it is not easily seen.

Blood supply: Epithelia are avascular. Nutritive materials and oxygen enters only by
diffusion through cells and intercellular substance. With few exceptions, epithelium of stria
vascularis of internal ear possess vascular supply.

Classification:

I. Simple Epithelium

1. Simple squamous epithelium (Pavemental epithelia): It consists of flat scale-like

or plate-like squamous cells separated by narrow intercellular spaces. The edge s of the cells
are usually serrated but may be smooth. The cells rest on the basement membrane. The
nucleus is situated in the centre of the cell and is spherical or oviod causing a bulging of
cytoplasm. On surface view, the cells appear as delicate mosaic which can be demonstrated
well by silver staining technique, wherein precipitation of silver occur in the intercellular
spaces. In histological sections, the cells are spindle shaped, thin at either ends and thicker
in the centre where the nucleus is situated.
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Distribution: 1. This epithelium lines the peritoneal, pleural and pericardial cavities. Here, it
is named as mesothelium. The mesothelium together with an underlying stratum of
connective tissue forms serous membranes viz., peritoneum, pleura and pericardium.
2. It lines the heart, blood vessels and lymph vessels; here it is named as endothelium.
3. It also lines the membranous labyrinth of the internal ear, Bowman’s capsule and alveoli of
the lungs.

2. Simple cuboidal epithelium: Consists of a single layer of more or less cubical

cells, rest upon a basement membrane. The cells are isodiametric, nucleus is single,
spherical and centrally placed. Found in many parts of kidney tubules, thyroid acinus, acinar
ducts, surface of ovary, most of the exocrine glands and mammary gland etc.

3. Simple columnar epithelium: Consists of a single layer of tall prismatic cells rest
upon the basement membrane. The cells are taller, column-like. Generally, the cells are
wider at the free end and narrow at the basal or attached ends. Possess oval nuclei, situated
at the basal part of the cell. Pyramidal epithelium of many glands is a modification of the
columnr epithelium.

Simple columnar epithelium is found in the stomach, intestine, interlobular ducts and
excretory ducts of many glands, in gall bladder, uterus, vas difference etc.

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 HISTOLOGY LECTURE NOTES

Goblet cell: A common variation of simple columnar epithelium is in the form of

goblet cells. These are unicellular mucous secreting glands found in the intestine and
respiratory passages. They become modified from the columnar cell by deposition of mucous
droplets. Mucus accumulation makes the cell to swell up and pushes the nucleus to the
periphery and is surrounded by a little cytoplas. On further expansion, the cell membrane
ruptures with escape of mucin. In ordinary preparations mucin is unstained. The cell after
rupture regenerates and the process is repeated.

4. Pseudo stratified epithelium:

In this type of epithelium, the nuclei, lies at different levels giving it a stratified
appearance. All the cells reach the basement membrane but not all of them extend to the
surface. Those reaching the surface are columnar with one or more processes extending to
the basement membrane. Between these are ovoid or spindle shaped cells, which also reach
the basement membrane. This is type of epithelium is usually ciliated (motile or stereo cilia),
the cilia being the present on the surface columnar cells. This epithelium occurs mainly in the
passages of the respiratory and male reproductive systems.

Fig.3.4 Pseudostraitified ciliated columnar epithelium

II. Stratified Epithelium
1. Stratified squamous epithelium: It is the main protective epithelium of body and has
several cell layers. The number of layers of the epithelium varies in different regions but
shape and the arrangement of the cells are characteristic. Two types of this epithelium are
recognized (i) Keratinizing and (ii) Non-keratinizing.

i. Stratified squamous - Keratinised: It consists of five layers of cells:

Deepest layer consists of columnar cells resting on an indistinct basement membrane

is referred as stratum basale or cylindricum. Above this, the cells become polyhedral and
19
 HISTOLOGY LECTURE NOTES

are usually larger than the basal cells. As the free surface is approached, the cells are more
flattened and at the surface they become squamous. The deeper cells of the basal
polyhedral layers (stratum germinativum) are young soft cells with large nuclei rich in
chromatin and a finely granular cytoplasm. The intercellular bridges are prominent, giving the
cell a prickly appearance (prickle cells) and hence this layer is called stratum spinosum.
Mitosis occur in these layers.

Above the stratum germinativum there is stratum granulosum consisting of 2 - 5

rows of flattened; rhomboid cells with basophilic kerato-hyalin granules in the cytoplasm.
Above this is the stratum lucidum consisting of a few row of flattened cells devoid of any
nuclei, appearing as a translucent, homogenous highly refractive band, in which cell
boundaries cannot be made out. The cytoplasm in these cells contains eleidin droplets.
Above the stratum lucidum is the most superficial layer the stratum corneum. The thickness
of this varies in different locations. It is composed of dead scales with no nucle us and is
filled with keratin. Stratified suqamous keratinized epithelium covers the entire body and the
orifices of the cavities opening upon it.

ii. Stratified squamous - Non-keratinised:

In non-keratinized epithelium, stratum granulosum, stratum lucidum and stratum

corneum are usually absent. The extent of the changes in the superficial cells varies with the
location and the environment of the epithelium. Its surface cells are non-nucleated, scale-like
and keratinized. Stratified suqamous non-keratinized epitheliumis located in the lining of

20
 HISTOLOGY LECTURE NOTES

oesophagus, mouth cavity, anal canal, anterior surface of the cornea, bulbar conjuctiva,
lacrimal canaliculi, vagina, glans penis and elsewhere.

The process of keratinization in epidermis is so important for bodily protection.

Keratinization process includes loss of nucleus, mitochondria and golgi apparatus along with
decreased lysosomal activiities and is probably due to insufficient nutrition of the upper cell
layers. The dead scale-like cells are constantly shed off to be replaced by cells from the
deeper strata. In man, this is slow and continuous process, in some of the lower vertebrates
(snakes), there is periodic shedding of the whole superficial layer of the epidermis.

Stratified columnar and stratified cuboidal epithelium: This type of epithelium is

comparatively rare. In the former, the most superficial layer of cells are columnar (e.g larger
excretory ducts of some glands, palpebral conjuctiva of the horse and dog). In stratified
cuboidal, the top cells are cuboidal. (e.g. ducts of sweat glands, cells lining antrum of the
ovarian follicles.

2. Transitional epithelium:

This epithelium is seen in the mucous membrane of the excretory passages of the
urinary system. The cells are often divided into three layers: the basal polyhedral cells, the
intermediate pear shaped or pyriform cells, with their broad, end towards the free surface
and flattened or umbrella-like squamous cells. This epithelium accommodates itself to cover
21
 HISTOLOGY LECTURE NOTES

larger or small areas. When the urinary baldder is empty, the cells are arranged in six or
more layers, while, distended, the cells are arranged in two or three layers only. The cells are
held together by viscous cement which permits gliding over of cells on one another.

Fig. 3.7 Transitional epithelium A. stretched state B. relaxed state

1. Superficial cells 2. Reduced number of layers
3. Umbrella cells 4. Pyriform cells
5. Basal cells
III. Others:

1. Glandular epithelium: refers to the lining and secretary cells of the glandular organs,
which are developed as down growths from the epithelial layer in the underlying connective
tissue, followed by structural modification and differentiation.

2. Neuro-epithelium: Epithelial cells associated with nerve endings. Restricted to certain

locations, which are concerned with the reception of sensory stimuli.

Mucous membrane (Mucosa): This lines all of the canals and cavities of body which
connect with the exterior that is the line alimentary tract, the respiratory passages and the
genitor-urinary tract. The essential parts of the mucous membrane are surface epithelium,
basement membrane and a stratum of connective tissue called the lamina propria. The
surface epithelium is of different types in the different systems and even in different parts of
the same system.

Epithelia generally possess a remarkable capacity for repair and regeneration after
injury. Certain epithelia, notably epidermis and intestinal epithelium, are continually recycled,
with new cells being created by mitotic activity while old cells are sloughed off (from the
surface of the epidermis or the tips of intestinal villi).

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 HISTOLOGY LECTURE NOTES

CONNECTIVE TISSUE

The various types of adult connective tissues in contrast to epithelium have relatively
a few cells and a large amount of extracellular matrix (ECM). But the proportion of these two
elements shows variations. ECM includes fibres and specialised proteins that constitutes the
ground substance.

In some type, especially loose connective tissue, cells are quite numerous. In other
types, they are few in number and tissue is composed almost entirely of closely packed
fibres. The type and the nature of the substance in which they are embedded (ground
substance) furnish the basis for the subdivision of adult connective tissue into three main
groups viz., connective tissue proper, cartilage and bone.

Two categories of connective tissue are recognized.

(1) Embryonal connective tissue and

(2) Adult connective tissue.

Embryonal connective tissue: are two varieties (a) Mesechyme and (b) Mucous.

Mesenchyme consists of a network of branching cells, in the mesh of which is a

homogenous intercellular fluid. The processes of a cell appear to anastomose with those of
other cells. As development progresses, wavy primitive fibres appear between branching
cells and the fluid matrix becomes viscid due to formation of mucoproteins. This
developmentally more mature tissue is mucous connective tissue which is widely distributed
in the embryo. The primitive fibres later become collagenous. Elastic fibres appear later.
Classical example of mucous connective tissue is umbilical cord and the term Wharton’s jelly
is applied to it in that location.

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HISTOLOGY LECTURE NOTES

Fig.3.8 Mesenchyme
1. Nuclei
2. Nucleolus
3. Cell processes
4. Scanty cytoplasm
5. Fluid filled spaces

Fig.3.9 Mucus connective tissue

1. Fibroblast
2. Fibres in jelly

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 HISTOLOGY LECTURE NOTES

LOOSE CONNECTIVE TISSUE

Loose or areolar connective tissue is very widely distributed in the body. It forms the
superficial and of the deep fascia; forms part of the frame work (stroma) of most of the
organs; or surrounds blood vessels and nerves and fills in any otherwise unoccupied spaces.
Areolar tissues contain cells, fibres and ground substance.

Cells: Cells constantly present are: 1.Fibroblasts 2.Macrophages or histiocytes

3.Mast cells, 4.Plasma cells, 5.Wandering cells. 6. Pigment cells and 7.Fat cells. Of these ,
fibroblast and histiocytes are the most numerous.

1. Fibroblasts: are larger flat branching cells with extensive processes which may
join the processes of other fibroblasts. The cell membrane is delicate and usually not seen.
The nucleus is spherical or oval and in ordinary preparations is lightly staine d. In section, the
fibroblast nuclei are usually shrunken and stain deeply with basic dyes. It contains dusk-like
chromatin and one or two nucleoli. The cytoblasm is homogenous or finely granular and
stains very lightly. Fibroblasts are present in all connective tissue such as tendon. They are
responsible for formation of fibres. The name fibroblasts instead of fibrocytes indicate that
after injury they are active and form new fibres. Fibrocytes on the other are inactive.
Their cell bodies are less irregular and nucleus is condensed to a long cylinder or flattened
oval. Chromatin is dense.

Fig.3.10 Areolar tissue

1. Fibroblast
2. Collagenous fibres
3. Mast cell
4. Elastic fibres
5. Macrophage
6. Intercellular
substance
7. Plasma cell
8. Fat cell
9. Blood vessel
10. Capillary

25
 HISTOLOGY LECTURE NOTES

2. Histiocytes or Macrophages: These are irregular cells with short processes.

The nucleus is smaller and darkly staining than fibroblasts: cytoplasm coarsely granular and
shows vacuoles. They are studied by vital staining. They are present in all fasciae and in the
stroma of organs. They also occur in sinusoids of liver, in lymphoid organs and in bone
marrow. They show amoeboid movement and phagocytosis in pathological conditions.
When activated, the histiocytes become larger and more rounded and the cytoplasm will be
filled with granules of ingested material. They form part of the macrophage system or
reticulo endothelial system.

3. Plasma cells: are comparatively rare in loose connective tissue, but are numerous
in the alimentary mucous membrane and the great omentum and also in pathological
conditions. They are smaller than macrophages, round or irregular in shape, with basic
staining homogenous cytoplasm. The nucleus is small and eccentrically placed. Large
granules of chromatin radially arranged in a regular manner give s a cart-wheel appearance
to the nucleus. There is a characteristic unstained or lightly stained area in the cyto plasm at
the side of the nucleus, where the cytoplasm is more abundant. They are derived from
lymphocyte-like cells and are active in forming antibodies.

4. Mast cells: Occur in most loose connective tissue especially along the course of
blood vessels. They are large oval or round cells with pale-staining nucleus and coarse
cytoplasmic granules which stain with certain basic dyes. They resemble basophils of blood
and contain an anticoagulant heparin, histamine and serotonin.

5. Wandering cells: are lymphocytes, eosinophils and neutrophils, migrated from

blood.

6. Pigment cells (Chromatophore): The cells of this tissue occur in the chorioid and
iris of the eye and in the corium of dark skinned animals. The cytoplasm is filled to a varying
degree with brown or black pigment which is usually melanin. They are specialized cells,
called as Melanocytes, irregular in outline with cytoplasmic processes.

7. Fat cells: The large ovoid or spherical cells consist of peripheral cytoplasmic
membrane enclosing a large fat droplet. The nucleus is flattened and surrounded by a small
amount of cytoplasm and is usually found pressed against the cell wall. In sections, when the
fat is dissolved out, the cells appear as empty rings or have a signet ring shape when the

26
 HISTOLOGY LECTURE NOTES

plane of section passes through the nucleus. This is so when they occur isolated or in small
groups. But in denser masses due to pressure of adjacent calls they become polyhedral.

Fig.3.11 Cells of loose connective tissue

Connective tissue fibres:

Three types of fibres are seen in adult connective tissue.

1. White or collagenous fibres: Collagen fibres are the most abundant type of fibre.
Under light microscope, they appear as wavy bundles; flexible structures of variable width
and length. They have high tensile strength and are non-extensile. Under EM, these fibre
bundles are madeup of extremely fine thread-like subunits called collagen fibrils, which lie
27
 HISTOLOGY LECTURE NOTES

parallel to one another giving the fibre a longitudinally striated appearance and are united by
a small amount of intercellular cement substance. Fibrillae do not branch. They stain with
acid dyes. Chemically they contain an albuminous substance called collagen which on
boiling yields gelatin. The fibres shrink slightly in water; swell enormously in dilute acids or
alkalies. In the dilute acids or alkali the fibres break up into fibrils, become individual as the
inter-fibrillar substance is dissolved. Gastric juice rapidly digests the fibres, but alkaline
trypsin has no effect on them. Collagen fibres are birefringent or aniostropic throughout t heir
length.

Based on the polymerization pattern, collagen fibres are categorised into 25 types. Of
which, type I and III collagen fibres are the most widely distributed types. Both are madeup
of trophocollagen fibrils.

2. Elastic or yellow fibres: are homogenous highly refractive fibres which are
thinner than the white fibres. They branch and anastomose freely forming networks. Smaller
fibres are round in cross section and larger are flat or polygonal. They are highly elastic.
Chemically they contain elastin which does not yield gelatin and has a remarkable resistance
to most agents. It is not affected by heat or cold water, by dilute acids or gastric juice. It is
however rapidly dissolved by pancreatin. The elastic fibres are lightly stained by ordin ary
methods, but deeply stained by orcein and resorcin fuchsin.

3. Reticular fibres: are smaller branching fibres which form net-like supporting frame
work or reticulum. They become black in silver impregnation methods. Hence they are also
called argyrophilic fibres. In areolar tissue, these reticular fibres are few. In glandular
organs and between muscle fibres they are numerous. In lymphoid organs and in red bone
marrow, the reticular fibres are associated with a special type of cells called reticular c ells.
Reticular fibres with reticular cells form a type of tissue called the reticular tissue. In other
situation reticular fibres have the same relationship to fibroblasts as do collagenous or elastic
fibres. Chemically though related to collagenous fibres the reticular fibres are not dissolved
by pepsin and on boiling yield reticulin and not gelatin.

Ground substance: It is the background material within which all other connective
tissue elements are embedded. It varies from a fluid-like to a gel-like state. It is one of the
two components of non-fibrillar intercellular substance. The other is the tissue fluid. In
ordinary connective tissue, the ground substance consists mainly of water whose major role
is to provide a route for communication and transport (by diffusion) between tissues. This
28
 HISTOLOGY LECTURE NOTES

water is stabilized by a complex of glycosaminoglycans (GAGs), proteoglycans, and

glycoproteins, all of which comprise only a small fraction of the weight of the ground
substance.

Ground substance may be highly modified in the special forms of connective tissue.

 In blood, the ground substance lacks stabilizing macromolecules. We call this free-
flowing ground substance as plasma.

 In skeletal tissue, the ground substance may become mineralized by deposition of

calcium salts. We call this rigid ground substance as bone.

 In cartilage, the ground substance is much more solid than in ordinary connective
tissue but still retains more resiliency than bone.

Functions of areolar tissue:

1. It binds structures or holds them in position.

2. It acts as padding and serves as a pathway for blood vessel and nerves. Nutrient
substances to body cells and metabolites from the cells traverse connective tissue.

3. Spread of localized infections is limited by connective tissue.

DENSE CONNECTIVE TISSUE

Here fibres are closely packed to form sheets, cords or bands (e.g dermis, capsules
of certain organs, aponeurosis, ligaments and tendon) based on the nature of arrangement
of the fibres, they are classified into two groups (1) irregularly arranged, (2) Regularly
arranged.

Dense irregularly arranged connective tissue occurs in the form of sheets. Main
component is coarse collagenous tissue but elastic and reticular fibres are also present. The
fibres interlace to form tough feltwork. Fibroblasts and macrophages are present
(e.g dermis of the skin, periosteum, perichondrium and capsules of organs).

Dense regularly arranged connective tissue occurs in the form of cord-like or band
like structures. Fibres are arranged parallel to one other (e.g tendons, ligaments and
aponeurosis).

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 HISTOLOGY LECTURE NOTES

1. Tendons are composed almost only of white fibrous tissue. Fibres are densely
packed in parallel bundles. Fibroblasts alone are present in small numbers. These are
called tendon cells. The tendon cells are quadrangular and so highly flattened that in profile
they appear as thin linear structures. They occur between tendon bundles around which they
have extensions.

2. Aponeuroses have same structure as tendons but are broad sheets. Fibres run in
superimposed layers those of one layer at an angle to adjacent layers. The layers may
interweave.

3. Ligaments are predominantly of white fibrous tissue but a few are composed
entirely of elastic fibres. The yellow elastic ligaments are formed of parallel cou rsing yellow
fibres, bound together by small amount of areolar tissue (e.g. ligamentum nuchae where
elastic fibres are very large and ligamentum flava).

SPECIAL CONNECTIVE TISSUE

Reticular Tissue :

It is composed of cells and reticular fibre network. The reticular tissue forms the frame
work of organs like lymphatic glands, spleen, red marrow of bones. The cells have stellate
shape with processes which extend in all directions and join with adjascent cells and
wrapped by reticular fibres.

The reticular cells line the lymph sinuses in lymphatic glands or blood sinusoids in
spleen, liver, red marrow etc. These cells are actively phagocytic and form part of the
reticulo-endothelial (RE) system.

Primitive or Non-phagocytic reticular cells: It appears likely that the reticular cells
may be of different types which cannot be distinguished by morphological means alone.
Some of them have retained their embryonic or developmental tendencies and in red marrow
give rise to erythrocytes and granulocytes; in lymphoid tissue to lymphocytes etc.

Adipose Tissue :

Fat is regarded as arising from specific cells called steatoblasts. Fat cells are found
isolated or in groups in all areolar connective tissue. But in adipose tissue, largest deposits of

30
 HISTOLOGY LECTURE NOTES

fat are found. Eg. subcutaneous connective tissue (panniculus adiposus), in the kidney
region, in mesenteries, in mediastinum and in the cervical, axillary and inguinal regions.

Fat is different from other connective tissues, in that the cells and not the intercellular
substance make up the bulk and determine the nature of the tissue. The large ovoid or
spherical cells consist of peripheral cytoplasmic membrane enclosing a large fat droplet. The
nucleus is flattened and surrounded by a small amount of cytoplasm and is usually found
pressed against the cell wall. In sections, when the fat is dissolved out, the cells appear as
empty rings or have a signet ring shape when the plane of section passes through the
nucleus. This is so when they occur isolated or in small groups. But in denser masses due
to pressure of adjacent cells they become polyhedral. Fat is usually arranged in groups or
lobules, each lobule being separated from the other by areolar connective tissue. Delicate
connective tissue passes between the cell and serves as bed for capillaries.

Chemically fat contains esters of glycerol and certain fatty acids. It is not soluble in
cold alcohol or water, but readily dissolves in xylol. That is why fat dissolves in ordinary
preparations. But by special methods fat can be stained (e.g. black by Osmic acid, red by
Sudan red III).

Brown fat tissue: This is different from the common or white fat tissue. In rats and
other rodents, it is highly developed and forms yellowish, lobated masses in certain parts of
the body - between the scapulae, on the neck, in the mediastinum, in the inguinal region and
elsewhere. Microscopically, latter name was given because the brown fat contains a pigment
which gives the tissue its colour. The fat cells assemble in groups separated by this network
of collagenous or reticular fibres and numerous capillaries. The microscopic structure
suggests an endocrine gland. While the common fat tissue loses or accumulates neutral fat
with changes in the nutritional condition of the animal, these factors do not seem to affect the
brown fat tissue. This tissue is believed to play a peculiar role during hibernation.

Pigmented Connective Tissue :

The cells of this tissue occur in chorioid, iris of the eye and in the corium of dark
skinned animals. The cytoplasm is filled to a varying degree with brown or black pigment
which is usually melanin. These specialized cells are called as melanocytes, irregular in
outline with cytoplasmic processes.

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 HISTOLOGY LECTURE NOTES

Blood and nerve supply of Connective tissue:

Connective tissue received its blood supply from the vessels which pass through
them and for which they form supporting bed. For dense connective tissue the supply is
scanty. Nerves are numerous in connective tissue. Some of them terminate in the tissue
itself, but other go to epithelium or muscle. Tendons, fibrous membranes and periarticular
connective tissue have special nerve endings.

Reticulo Endothelial System

This name is given to the system more on physiological and pathological

consideration than on anatomical structure. The cells of this system do not form true
endothelium.

All highly phagocytic cells of the body, except leucocytes belongs to this system.
The only method of identification of these cells is by injection of non -toxic, inert particulate
matter like carbon particles or Indian ink into the living animal. Injection of trypan blue or
lithium carmine into living animal is called vital or intra-vital staining. The macrophages
engulf the particulate matter like carbon particles. After vital staining the cells containing the
dye are found in the cells in loose connective tissue, in the reticular tissue or in the blood
sinuses of certain organs. In the connective tissue they correspond to the hist iocytes.
Kupffer cells of liver, reticular cells of lymph nodes, spleen and bone-marrow, cells lining the
sinusoids of spleen and microglia cell of the nervous system are examples of this system.

CARTILAGE

Cartilage consists of the cells termed as chondrocytes, matrix (ground substance)

and fibres. The matrix is solid and has marked elastic properties. It withstands considerable
pressure and tension.

According to the nature and visibility of fibrillar elements, cartilage is subdivided into
three varieties: 1.hyaline, 2. elastic, 3. white fibrous. Of these, hyaline cartilage is the most
widely distributed type of cartilage.

Hyaline cartilage:

This appears as a bluish-white translucent mass when fresh. It forms the articular
cartilage of joints, costal cartilages and cartilages of nose, larynx trachea and bronchi.

32
 HISTOLOGY LECTURE NOTES

In the foetus, nearly entire skeleton is first laid down as hyaline cartilage and later replaced
by osseous tissue in the formation of bone. The tissue is organized into plate-like masses
which with the exception of articular cartilages, are invested by a fibrous membrane, the
perichondrium. Each plate consists of cells and a homogenous matrix, the connective tissue
fibrils being masked by ground substance. There is neither blood supply nor nerve supply for
the cartilage.

Cartilage cells (chondrocytes) are situated in smooth-walled empty spaces called

lacunae. Each cell has a large central spherical nucleus with one or more nucleoli. The
cytoplasm is finely granular and contains fat, glycogen and occasionally pigment. In life ,
chondrocytes fill up the lacunae, but after fixation in ordinary methods, fat and glycogen are
dissolved and the shrunken cells are separated by spaces from the walls of the lacunae, give
the cells a branched appearance.

In the centre, the cells are arranged in isogenous groups, each group representing
the offspring of one parent cartilage cell. The cells are spherical or ovoid, flattened on
adjacent sides, their long axes directed radially towards the surface of the plate. Whereas, at
the periphery, the cells become flattened loose their definite grouping and in the
subperichondrial layers they appear as rows of narrow elongated cells with their long axes
parallel to the surface. Cartilage here merges insensibly with the fibrous connective tissue of
the perichondrium.

Ground substance: It appears homogenous in fresh condition or in ordinary

preparations. Only the walls of the lacunae stand out as mere refractive rings intensely
staining with basic dyes. These rings are called capsules. These are not of the nature of
cell membranes but represent condensation of the matrix surrounding the cells. The ground
substance or matrix masks the fine collagenous fibres, which are present in the homogenous
intercellular substance because both have the same refractive index. The ground substance
is strongly metachromatic with toluidine blue and is slightly basophilic but the superichondrial
layer is acidophilic.

Elastic cartilage:

In this, elastic fibres are present and embedded in the matrix. These fibres branch
and run in all directions and anastomose to form a dense network. In the peripheral layers,
the fibres are thin and the network wide meshed. In the deeper portions, they are thicker and

33
 HISTOLOGY LECTURE NOTES

more closely packed. Elastic cartilage is present in the external ear, the eustachian tube, the
epiglottis and in a part of aretynoid cartilages.

White-fibro cartilage :

This is a combination of dense collagenous fibres and cartilage cells. The cells are
surrounded by capsules which in turn are surrounded by variable amounts of matri x.
Frequently, the cells enclosed in capsules, lie in rows between which are dense wavy
bundles of collagenous fibres. Fibrous cartilage occurs in intervertebral, discs, interarticular
cartilage, the glenoid and cotyloid ligaments and the ligamentum teres of the femur. It is
found at certain attachment of the ligaments or tendons to bone.

Fig.3.12 Hyaline cartilage (T.S)

Fig.3.13 Elastic cartilage (T.S)
1. Perichondrium
1. Lacuna
2. Fibrocyte
2. Chondrocyte
3. Collagenous fibres
3. Central dense elastic fibres
4. Blood vessels
4. Peripheral less dense elastic fibres
5. Mesenchymal cells
6. Chondroblasts
7. Chondrocyte
8. Capsule
9. Lacuna
10. Cell nest 34
11. Matrix
 HISTOLOGY LECTURE NOTES

Fig.3.14 Whitefibro cartilage (T.S)

1. Chondrocyte s
2. Collagen fibres
3. Matrix

Perichondrium: Surface of cartilage is invested by a connective tissue membrane

called perichondrium which consists of an inner chondrogenic and outer fibrous layer.

BONE (OSSEOUS TISSUE)

Bone or osseous tissue consists of cells, collagenous fibres, and an intercellular

matrix of organic substances. The organic matrix is chiefly made up of osseo mucoid and in
this matrix, collagen fibres are embedded and the fibrils form a major portion of the
intercellular substance. The inorganic matter is composed of calcium phosphate, calcium
carbonate and small amount of other minerals. The minerals are present in the form of
crystals of hydroxyaptite embedded in the matrix and are arranged in a regular manner in
relation to the collagen fibres.

The bone cells or osteocytes are irregularly oval cells with fine cytoplasmic
processes, a large oval nucleus and faintly basophilic cytoplasm. These cells occupy spaces

35
 HISTOLOGY LECTURE NOTES

called lacunae in the solid intercellular substance and numerous canaliculi radiate from the
lacunae and are occupied by the cytoplasmic processes of osteocytes.

The intercellular substances of bone are in the form of thin plates or lamellae
between or within which are the lacunae and canaliculi occupied by osteocytes and their
processes. The arrangement of these lamellae is different in compact and cancellated bones.

Fig.3.15 Different bone cells

1. Osteoblasts
2. Osteocytes
3. Osteoclast
4. Bone matrix
5. Lacuna
6. Canaliculi

Organisation of bone tissue :

Grossly two types of bone may be distinguished the spongy or cancellous and the
dense or compact. The spongy and compact varieties do not represent histologically distinct
types of osseous tissues, but differ only in the degree of porosity. In the spongy bone, the
space is large and the bony matter reduced to a network of slender bars of bone. In compact
bone, the bony tissue is abundant and more densely packed. The histological character of
the cells and intercellular substance are the same in both.

36
 HISTOLOGY LECTURE NOTES

In all bones, there is an outer layer of compact bone enclosing the cancellated bone
and within the cancellated bone is the bulk of the bone, except in the diaphysis of long bones
and certain flat bones of the skull.

Compact bone:

This is composed of numerous systems of concentrically arranged bony lamellae,

around a central canal, the Haversian canal, which run longitudinally and encloses blood
vessels. Each concentric system is known as a Haversian system or Osteone. Haversian
canals of adjacent systems are inter connected by transverse channels and volkmann’s
canals connect haversian canals with periosteal and endosteal surfaces.

In a cross section of the bone, the Haversian canals are seen to be surrounded by a
varying number (8 to 15) of concentric lamellae. These concentric lamellae and the central
canal constitute a Haversian system or Osteone. In the periphery, the lamellae are not
concentric but run parallel with the surface and form a relatively thin outer layer of the bone,
the outer circumferential or general lamellae. Similarly arranged inner circumferentia l
lamellae separate the Haversian systems from the marrow cavity. Finally, the intervals
between the Haversian lamellae are occupied by more irregular layers of bone which
constitute the intertitial lamellae. Adjacent lamellar systems are as a rule sharpl y delimited
from each other by a dark staining thin layer of modified matrix (cement membrane, cement
line).

Cancellated bone :

This consists of a network of columns of lamellae enclosing spaces which contain

bone marrow. There are no Haversian systems.

Periosteum and Endosteum:

Periosteum is a fibrous membrane investing the bones except at their articular

surfaces. Periosteum consists of two layers, the outer of which is composed of course fibrous
connective tissue containing few cells but numerous blood vessels and nerves. The inner
layer is more cellular and less vascular and contains many elastic fibres. In the growing
bones, the inner layer of periosteum is osteogenic and in the adult, this layer is converted
into a row of flattened cells. Periosteum serves as a supporting bed for blood vessels and
nerves going to bone and for anchorage of tendons and ligaments. Some of the periosteal

37
 HISTOLOGY LECTURE NOTES

fibres pass into the bone, either obliquely or at right angles to the long axis of bone and are
termed as perforating fibres of Sharpy.

Endosteum lines the surface of the cavities within a bone (Haversian canals) and
also the surfaces of trabeculae in the marrow cavity. The cells of endosteal layer are like
those of the periosteum and rest on a thin layer of connective tissue.

Fig.3.16 Compact bone organization

1. Fibrous layer of periosteum 2. Sharpey’s fibres
3. Osteogenic layer of periosteum 4. Concentric lamellae
5. Canaliculi 6. Interstitial lamellae
7. Lymphatic vessel 8. Artery
9. Nerve 10. Vein
11. Osteone 12. Osteocyte
13. Medullary membrane 14. Inner circumferential lamellae
15. Osteone with osteocytes, lacuna and canaliculi
16. Lacuna with canaliculi 17. Haversian canal
18. Outer circumferential lamellae 38
 HISTOLOGY LECTURE NOTES

Fig.3.17. Fibre of Sharpey

(C.S)
1. Osteocytes
2. Bone matrix
3. Fibroblasts
4. Sharpey’s fibres
5. Periosteum

Bone marrow

This is a soft tissue which occupies the medullary cavity of the long bones and the
space between trabeculae of spongy bone. It consists of a delicate reticular connective
tissue in the meshes of which are many kinds of cells. Two varieties of marrow are seen, red
and yellow.

Red marrow: It is the only type found in foetal and young bones, but in the adult it is
restricted to the vertebrae, sternum, ribs, cranial bones and the epiphyses of long bones.
It is the chief blood forming organ of the adult body being the normal source of the red blood
corpuscles and granular leucocytes.

Yellow marrow: Consists of mainly fat cells which have gradually replaced the other
marrow elements.

Blood vessels and Nerves of the bone : Bone is richly supplied with blood vessels
and nerves which pass into it form the periosteum. An artery, the medullary artery runs
through the nutrient canal, through the nutrient foramen into the marrow cavity. In its course
through compact bone, the branches of the medullary artery communicate with the vessels of
the Haversian canal. The medullary artery divides into ascending and descending branches

39
 HISTOLOGY LECTURE NOTES

to supply all part of marrow and terminates in a network of sinusoidal capillaries. Veins
arising from these accompany the medullary artery and communicate with veins of the
Haversian canals. Others enter the compact bone through Volkmann’s canals and run in the
Haversian canals. They enter marrow cavity and anastomose by branches with the
capillaries of the medullary artery.

Nerves enter through Volkman’s canals and ramify in the Haversian canals and the
marrow. Pacinian corpuscles occur in the periosteum, whereas osseous tissue is insensitive.

BLOOD AND LYMPH

Blood is the connective tissue consisting of free cells and a fluid intercellular
substance or plasma. The structural elements of Mammalian blood include red blood
corpuscles (erythrocytes) white blood corpuscles (leucocytes) and blood platelets.

ERYTHROCYTES (Red Blood Corpuscles)

They are highly differentiated and specialized for the function of transporting oxygen.
In the lower vertebrates, the erythrocyte is nucleated but in mammals, it loses it nucleus,
Golgi apparatus, mitochondria and centroiles before entering blood stream. The erythrocytes
are acidophilic biconcave discs and round in all mammals except in camel and llama where
they are elliptical. (Birds, reptiles and fishes have elliptical nucleated erythrocytes).
On surface view, the erythrocyte is circular in outline and the central depression appears as
a lighter area depending on the focus.

Histologically, the erythrocytes appear entirely homogenous. The covering is a

delicate plasma membrane formed by a condensation of the lipo -protein complex of the
corpuscular substance. The contents of the corpuscle are present as a relatively fluid
colloidal mixture.

Chemically the erythrocyte consists of a protein and lipoid colloidal complex, of which
the most important element is hemoglobin. Hemoglobin is the pigment which gives the red
colour to the erythrocytes..

The average diameter of the erythrocyte is as follows in microns: Elephant 9.0; man
7.3; rabbit 6.5; cow 6.1; cat 6.0; horse 5.7; sheep 5.0; goat 3.7; musk ox 2.5.

40
 HISTOLOGY LECTURE NOTES

Number (in millions per cubic m.m.) Goat 17.3, Sheep 11.5, cat 7.2, Horse 7, Dog
6.1, Ox 6, Man 5; Guinea pig 5; Rabbit 4.6, Chicken 2.9.

The number, the size and shape of the erythrocytes are correlated to the metabolic
activity of the different animal groups. High metabolic activity is correlated with small size an d
circular shape and large number of cells conversely; lower metabolic rate is correlated with
larger size and small number of cells.

LEUCOCYTES (WHITE BLOOD CORPUSCLES)

These are nucleated, (true cells) and may be divided into non-granular
(agranulocytes) and granular (granulocytes) varieties. The cytoplasm of granular leucocytes
shows numerous granules. In many, these granules are fine and slightly refractive but in
others they are coarse and highly refractive.

Total number of leucocytes in thousand Per cubic mm Chicken 19.8; cat 17.2; swine
14.9; goat and guinea pig 12; dog 11.3; sheep 9.2; horse 9; man 9; ox 8; rabbit 8;

Granulocytes:

These are divided into three classes according to the nature of the staining reaction
of the granules present as neutrophils, eosinophils and basophils.

Neutrophils: They are large cells 8 to 12 microns in size. The nucleus shows 3 to 5
lobes connected by fine strands and is in the form of 2 or horse shoe. Granules are
neutrophilic as they have affinity towards the neutral stain of the Romanowsky compound
and take a light purple stain. In the rabbit, guinea pig and chicken they take acid stain. In
different species, based on staining reaction, these cells are also called heterophils.

They are highly phagocytic and show great amoeboid motility. After the migration
from blood stream, the neutrophils phagocytose bacteria or other small particles. They have
been cells that engulf larger particles. The neutrophils also elaborate powerful proteolytic
enzymes which may act within the cell as in the digestion of phagocytosed bodies liberated
and act outside the cell body. Chemotactically attracted by devitalized tissue bacteria or
other foreign bodies the cells reach these places by amoeboid movement and they liberate
the proteolytic enzymes either by secretion or most commonly by disintegration and rupture
of the leucocytes themselves. These cells form the so called pus corpuscles.

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 HISTOLOGY LECTURE NOTES

Number of neutrophils in percentage of total number of leucocytes; Dog 74; man 72;
Cat 59; horse 58; swine 53; guinea pig and rabbit 40; goat 35; ox 30; chicken 24.4; sheep 24.
In man, dog cat, horse and pig, the neutrophils are in majority. Whereas in ruminants,
chicken and laboratory rodent, the lymphocytes are in majority.

Eosinophils: These are larger than neutrophils and the size is about 10 to 14
microns. It contains spherical and highly refractive granules which stain with eosin and other
acid dyes. The nucleus generally has only two lobes. The granules are especially large in
horse. They show amoeboid movement but are not actively phagocytic. Their number is
greatly increased in allergic states, bronchial asthma, skin diseases and parasitic
infestations.

Number of eosinophils in percentage of total number of leucocytes Ox.10; guinea pig

8; horse 7; goat 5.8; cat 5; sheep 4, swine 4; dog 2, chicken 1.9; rabbit 1.5.

Basophils: These are rare in adult blood being only 0.5 percent of leucocytes except
in the chicken (3 percent). Their size is 8 to 10 microns. The nucleus is large irregularly
polymorphous. The coarse granules are basophilic, intermediate in size between eosinophil
and neutrophil types and are soluble in water. They resemble the mast cells of connective
tissue.

Non-granular leucocytes or agranulocytes:

These include the lymphocytes which are small about the size of erythrocytes, and a
group of largest cells, the monocytes which have more cytoplasm and a more indented
nucleus.

Lymphocytes: The size of the small lymphocyte varies from 6 to 9 microns majority
being 7 to 8 microns. The nucleus is large and stains intensely. The basophilic cytoplasm
forms a narrow rim around the nucleus. The large lymphocytes measures from 10 to 12
microns. The larger lymphocytes also have a deeply stained spherical nucleus and a
basophilic cytoplasm, but there is greater proportion of cytoplasm than in the small
lymphocyte. The basophilia of the cytoplasm in both is due to diffuse distribution of free
ribosome or RNA granules.

The lymphocytes migrate through walls of capillaries into the connective tissue where
they are believed to perform a number of functions. They are believed to act as precursors

42
 HISTOLOGY LECTURE NOTES

of monocytes and macrophages in the connective tissue. Following a primary antigenic

stimulation, the lymphocytes give rise to a line of cells which d ifferentiate into plasma cells,
which produce the specific antibodies. They probably have a detoxifying action and are
concerned in protection against foreign substance including response against grafts of
foreign tissue.

Number of lymphocytes in percentage of the total number of leucocytes:- Sheep 68;

chicken 62; goat 57; rabbit 55; ox 52; guinea pig 45; swine 38; horse 28; man and dog 20.

Monocytes: These are large cells, size varying from 12 to 15 microns, but they may
occasionally be up to 20 microns. The nucleus is ovoid, kidney or horse-shoe shaped and
eccentrically placed. It stains less intensely than lymphocytes. Cytoplasm is basophilic and
abundant. They migrate through the capillary wall into the surrounding connective tissue,
become actively phagocytic and transform into macrophages, indistinguishable from the
macrophages present in the connective tissue. They serve to combat certain types of
bacterial infection like tuberculosis.

Number of monocytes in percentage of total number of leucocytes: Chicken 9.4; ox 7;

guinea pig 6; horse 5; man 5; swine 4; sheep 3; rabbit and cat 2.5; goat 2.2.

Blood Platelets: They are colorless about 3 microns biconvex discs but appear
spindle shaped when seen sideways. They have no nucleus. They show a central granular
mass which stains deeply with basic stains called the chromatomere and a peripheral hyaline
lightly stained zone called the hyalomere. In mammals they are not cells but are only
fragments of cells derived from the giant cells in red marrow. Birds an d other lower
vertebrates have true thrombocytes which are nucleated and have basophilic cytoplasm.
The platelets play an important role in coagulation of blood.

LYMPH

Lymph consists of a fluid, in which various corpuscular elements are suspended.

Red blood corpuscles and platelets are absent. The chief cellular elements are lymphocytes
with few granulocytes.

RED MARROW (Myeloid tissue): It is composed of stroma of reticular cells and

fibres with free cells located in the meshwork. The stroma also shows varying number of fat
cells. Majority of free cells in bone marrow are the developmental stages of the granulocytes

43
 HISTOLOGY LECTURE NOTES

and a few lymphocytes, monocytes and plasma cells. Sinusoids are numerous and form
anastomosing networks.

Haemocytoblasts: They are large cells up to 15 microns in size. The nucleus is large
and rounded and the cytoplasm is deeply basophilic. They constitute normally 0.3 to 0.5
percent of the cells of marrow.

Erythroblasts: These are the precursors of erythrocytes and undergo a series of

divisions and transformation before the erythrocytes is formed. The important stages in
erythrocyte formation are (1) Proerythroblasts or Basophilic erythroblast
(2) Polychromatophilic erythroblasts (3) Normoblast and (4) Erythrocyte.
Proerythroblasts (Basophilic erythroblasts): These are large round cells with
spherical nucleus containing angular particles or chromatin and the cytoplasm is
homogenous and is intensely basophilic.
Erythroblasts: These arise by mitotic division of proerythroblasts. There is an
increase in the haemoglobin in successive stages so that erythroblasts at successive stages
show less of basophilia and more of acidophilia. Hence these cells are called poly -
chromatophilic erythroblasts. Chromatin becomes more regularly arranged. The cells are
smaller than proerythroblasts and larger than normoblasts.
Normoblasts: These are derived from polychromatophilic erythroblasts after a
number of divisions. The cytoplasm has more haemoglobin and stains like erythrocytes.
The cells are slightly larger than erythrocytes. Their nucleus is smaller and dense in
chromatin and stain intensely. The normoblasts extrude their nuclei to become erythrocytes.
The youngest erythrocyte shows a delicate reticulum and so it is called reticulocyte.
The reticulum disappears before reticulocytes leave the marrow.
Myelocytes: These differentiate from myeloblasts. These constitute 12 percent of
the marrow cells. These cells show an increase in the granules and in crease in the density
of chromatin of the nuleus. The earliest myelocytes are known as Promyelocytes. They are
large cells with rounded or oval nucleus and basophilic cytoplasm. The cytoplasm contains a
few granules of one of the specific types (neutrophilic or acidophilic) but because of the few
granules promyelocytes are not classified into three groups. The promyelocytes proliferate
and differentiate into myelocytes proper. The cells become smaller in size show an increase
in the number of granules, decrease in the basophilia of cytoplasm and increase in the
density of the chromatin of the nucleus. Neutrophilic, esoinophilic and basophilic myelocytes
are distinguished. The myelocytes after a number of generations differentiate into

44
 HISTOLOGY LECTURE NOTES

metamyelocytes. They have an indented or kidney shaped nucleus and the cytoplasm
showing numerous specific granules of each type. The metamyelocytes differentiate into
mature leucocytes, which enter the blood stream.
Megakaryocyte: These are also called as giant cells and are large (30-100 microns)
rounded cells but the cytoplasm may show irregular processes. The nucleus consists of
many lobes connected by short stalks. Processes of cytoplasm of their cell extend through
the walls of sinusoids and by constriction and segmentation.

MUSCLE

Three types of muscle viz., smooth, skeletal and cardiac are distinguishable
histologically. Smooth or non-striated muscle lacks the cross striations seen in other two
muscles. The smooth muscle is involuntary, being especially associated with viscera which
are not under the voluntary control. Skeletal muscle is striated and voluntary making up the
bulky muscles of the body. Cardiac or heart muscle is striated but involuntary.

SMOOTH MUSCLE

Consists of fusiform or spindle shaped cell 15 to 500 microns in length with abundant
cytoplasm in whose central thickest position the nucleus lies. The nucleus is oval elongated
or rod-shaped. The nuclei of contracted muscle cells usually have a folded outline.
The cytoplasm contains the usual organoids and appears more or less homogenous in
routine sections. By treatment with dilute nitric acid, longitudinally arranged myofibrils can be
made out. The cytoplasm in routine preparations takes the eosin stain. A plasma membrane,
similar to those in other cells is present.

As a rule the cells are gathered into dense sheets or bands, but they may occur as
isolated units scattered among the connective fibres in a tissue like the dartos of the scrotum.
Within the bands the cells are parallel to each other but are packed together so that the
narrower position of one lies against the wider portions of its neighbors. Delicate reticular
and elastic fibres surround individual cells in a bundle. Between larger bundles there is
collagenous tissue.

The smooth muscle cells increase in the pregnant uterus where the length may
exceed half m.m. (500 microns). Smooth muscle is not richly supplied with blood vessels.
Capillaries are seen in between groups of cells. There are nerve endings in smooth mus cle

45
 HISTOLOGY LECTURE NOTES

but motor termination for every cell is not seen. The contraction of smooth muscle is slow
and sustained, in contrast to the skeletal muscle which contracts rapidly.

Smooth muscle is found in the wall of the alimentary canal from the stomach to anus,
gall bladder, trachea, bronchial tree, ureter, bladder, prostate and Cowper’s gland , oviduct,
uterus, vagina, blood vessels and larger lymphatic vessels, spleen and lymph nodes. Smooth
muscle occurs in skin in connection with hair follicles. In the eye, it is found in the Iris and
ciliary body.

Fig.3.18Smooth muscle
A. Visceral smooth
muscle tissue
B. Multiunit smooth
muscle tissue
1. Autonomic
neurons
2. Nucleus
3. Muscle fibers

SKELETAL MUSCLE

The independent units of skeletal muscle are called as muscle fibres. The fibres have
many nuclei and are larger than most cells. Fibres are grouped together into bundles of
fasciculi. Larger muscles are composed of many fasciculi.

The skeletal muscle fibres are elongated and cylindrical; 1.5 cm in length and
10-15 microns in diameter. The thickness varies with species, individual stage of nutrition,
age, size and especially function of muscle. With growth or exercise the size increases.

Surrounding the fibre is a covering or sheath called Sarcolemma. This sheath is

elastic, transparent, homogenous and difficult to see except when broken in the fresh

46
 HISTOLOGY LECTURE NOTES

preparations. It can be seen in stained longitudinal sections. Inside the sarcolemma, are the
nuclei and a cross striated substance composed principally of myofibrils. Surrounding fibrillae
is sarcoplasm which corresponds to cytoplasm of other cells. The skeletal muscle fibre has
many nuclei, several hundred appearing within a fibre of average size. The nuclei are directly
under the sarcolemma. The nuclei are flattened and oval. They show a loose network of
chromatin.

Fig.3.19A Skeletal muscle fibre (molecular level)

The skeletal muscle fibre shows a characteristic striated appearance du e to regular

alternation of dark and light stripes or striations across. They are produced by alternating
light and dark segments of longitudinally arranged elements, by myofibrillae. The latter are
1.2 microns in diameter. They are visible within the fibres of teased fresh muscle as well as
in fixed preparations. Similar segment of adjacent fibrillae are placed side by side in an intact
fibre to form characteristic cross striations seen in the fibre as a whole.

47
 HISTOLOGY LECTURE NOTES

In a cross section of a fibre the cut ends of myofibrils are visible as definite areas with
individual size and shape. They are separated from each other by narrow spaces which
represent the sarcoplasm. In routine preparations the fibrils appear to be arranged in
irregular groups known as Cohanheimsfields.

In the cross striations of the fibrillae the dark band is called the A or Q band
(anisotropic). The light band is designated I or J band (isotropic). Each of these bands is
bisected by a narrow line, that in the 1 band stains deeply and is design ated ‘Z” line and the
line bisecting the A (q) band is pale deeply and is called H zone. The Z line is also called
Krause’s membrane. The portion of a fibril between the two successive ‘Z’ line is called a
sarcomere. In the relaxed condition, the sarcomere is 3 microns.

Each myofibril is made up of thin, thread like elements myofilaments. Two types of
myofilaments have been observed. One is thick and extends from one end of the A band to
the other. These are myosin filaments. The thin filaments from either side of the ‘Z’ line
extend across the adjacent I band into the ‘A’ band as far as the ‘H’ zone. These are made
up of the actin and tropomyosin. Thus the cross striations seen with the light microscope are
related to the distribution of myofilaments which can be seen only under electron
microscope.

Sarcoplasm fills in the spaces between myofibrils is clear cytoplasmic ground

substance. It contains Golgi apparatus, mitochondria, endoplasmic reticulum and has few
ribosomes. It also contains glycogen and fat droplets. Fibres rich in sarcoplasm appear
darker with the naked eye or low magnifications. These are called dark fibres or red fibres.
Those poor in sacroplasm are light fibres. Both kinds are intermingled in the muscles of
mammals.

Changes during contraction: During contraction the fibre as a whole becomes

shorter and broader, when it contracts. Each sarcomere also becomes are stimulated to
contract by about 50 per cent. The H zone of A band usually disappears at the same time.
The total length of A band remains constant usually during contraction and relaxation. It is
believed that during contraction the thin actin filaments slide over the thick myosin filaments,
the actin filaments being more or less pulled into the “A” band or that the actin fi laments fold
or coil up during contraction. During contraction an acto-myosin complex is formed.

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 HISTOLOGY LECTURE NOTES

Fig.3.19B Changes in Skeletal muscle fibre during function

Muscle tendon attachment: At the ends of a skeletal muscle the fibres are attached
securely either to tendon or periosteum by some fibrous connective tissue structure. Muscle
fibrillae end abruptly and are not continuous with those of tendon or other connective tissue
structure. But the connective tissue of muscle (see below) is continuous with the connective
tissue of the tendon or other connective tissue structures to which muscles are attached.

Connective tissue of muscle: Surrounding each muscle fibre is a network of fine

reticular fibres the endomysium. Groups of muscles fibres are surrounded by collagenous

49
 HISTOLOGY LECTURE NOTES

connective tissue called perimysium. Thus a dozen or more muscle fibres are completely
surrounded by perimysium to form fasiculi or muscle. Fasciculi are in turn bound into larger
and larger orders of bundles by epimysium and the entire muscle has its outer investment
the deep fascia of gross anatomy. Endomysium, perimysium and epimysium are continuous
with one another in the order mentioned blood vessels and nerves ramify through the
connective tissue.

Blood and nerve supply: The larger branches of arteries penetrate the muscle by
following laminae of perimysium. The arterioles penetrate the fasciculi capillary supply is
very rich. Every skeletal muscle fibre receives at least one motor nerve ending from different
nerves. The sensory or afferent fibres are associated with specialized endings called
neuromuscular spindles (Refer Nervous tissue).

CARDIAC MUSCLE

Myocardium or heart muscle is composed of an interlacing network of muscle fibres

(cells). The muscle net is irregularly and incompletely divided into bundles and laminae which
wind about the heart in the form of spirals. The fibres within a bundle are mostly parallel but
the bundles themselves course in different directions so that a section of myocardium shows
fibres cut in longitudinal, transverse and oblique planes.

The muscle substance is composed of fibres or cells which show cross striations like
skeletal muscle and are 9 to 20 microns in diameter, connected with each other by
anastomosing branches. The myofibrils resemble that of skeletal muscle but the cross
striations are closer together and not very distinct. These fibrillae are thicker and more
closely packed together at the periphery of the fibre but may be so fine and sparse in the
centre that there appears to be an axial core of sarcoplasm. In cross section the
arrangement of cut ends of fibrillae frequently suggests short parallel bands or the spokes of
a wheel Nuclei are in the interior near the central axis. They are oval and quite large. There
is an accumulation of sacroplasm around the nucleus which contains mitochondria, fat
droplets etc. A sarcolemma or plasmalemma is presented similar to that of the skeletal
muscle.

Cardiac muscle also shows intensely staining transverse bands, at certain levels and
these are known as intercalated discs. These are 0.5 to 1 micron thick that is less than a
cross striation or sarcomere. They are strongly refractile in fresh muscle, but deeply stained

50
 HISTOLOGY LECTURE NOTES

in fixed material. They run in a straight line across but frequently are irregular or broken into
“step formations”. They are the cell membranes of adjacent cells meeting each other at cell
junctions.

Connective tissue of cardiac muscle: A network of reticular fibres and fine

collagenous fibres surrounds each muscle fibre. It corresponds to endomysium of skeletal
muscle. Between the bundles of muscle fibres there are coarser collagenous fibres and
elastic fibres. These regions corrsponds to the perimysium of skeletal muscle.

Blood vessels and nerve supply: An extensive plexus of blood and lymph
capillaries is found in the connective network surrounding each muscle fibre. Branches of
sympathetic and parasympathetic (vagus) nerves terminate in fine free endings on the
muscle fibres.

Conducting system: Some of the cardiac muscle fibres are modified to form the
impulse conducting system, whose function is to regulate successive contractions of atria
and ventricles. It extends from the right atrium into the ventricles and is known as the atrio -
ventricular bundle of his. The AV bundle and its branches consist of modified cardiac muscle
fibres called purkinje fibres.

They are distinguished form cardiac muscle fibres by (1) the reduced number of
myofibril which are restricted to the periphery of the fibre (2) greater amount of sarcoplasm
(3) more rounded nuclei, which occur in groups of two or more innermost of the cells.

Regeneration of muscle: With slight injury the smooth and skeletal muscle fibres
may proliferate but a larger defect of the muscular tissue is made good only by a connective
tissue scar. The cardiac muscle has little regenerative capacity and healing is by formation of
scar tissue.

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 HISTOLOGY LECTURE NOTES

NERVOUS TISSUE

The nervous tissue is specialized to receive stimuli from the environment to transform
them into nerve impulses and to transmit them to the nerve centers, from where appropriate
response is transmitted to another organ or part of the body which reacts to the response of
the original stimulus.

Nervous tissue consists of neurons and supportive cells called neuroglia. Nervous
tissue constitutes the nervous system which is divided into the central nervous system and
the peripheral nervous system. The central nervous system (CNS) includes the brain and
spinal cord. The peripheral nervous system (PNS) consists of cranial and spinal nerves,
including associated nerve roots and ganglia. Nerves and ganglia that innervate viscera are
designated the autonomic nervous system (ANS).

NEURON

The neuron is the structural and functional unit of the nervous system. Although
neurons show the greatest variation in size and shape of any group of cells in the body, they
can be grouped into three general categories.
Morphologically, neurons feature elongated processes, termed axons and dendrites,
that extend variable distances from the cell body (perikaryon). Metabolically, neurons are
actively involved in maintaining their structural integrity and in synthesizing, packaging,
transporting, and releasing secretory products. Neurons specialize in excitability and they
communicate by releasing chemical agents (neurotransmitters, neuromodulators, or
neurohormones).

Neuronal structure:

Neuron consists of Cell body (Perikaryon) consisting of nucleus, Neuronal

processes namely axon and dendrites.

Cell body

The cell body (perikaryon; soma) of a neuron consists of the nucleus plus the
surrounding cytoplasm and plasma membrane. In routinely stained histologic sections, the
cell body is the most identifiable feature of a neuron. Cellular constituents are synthesized
within the cell body and then flow distally into neuronal processes (axon and dendrites).

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 HISTOLOGY LECTURE NOTES

Cell bodies ranging from less than 10 micron to more than 100 micron in diameter.
Cell body size is proportional to neuronal total volume, although the cell body itself
represents a minor portion of the total volume.

Nucleus is pale staining and vesicular but the nuclear membrane stains dark with
haematoxylin- chromatin is fine and dispersed. Nucleolus is single larger very prominent and
may be basophilic or acidophilic. In sympathetic ganglia, the mitochondria, Golgi apparatus
chromophil substance or Nissl bodies and neurofibrils. The neurofibrillae are fine cytoplasmic
fibrils which form bundles are interlace in the cell body but are parallel in axon and dendrites,
demonstrable by special techniques Neuroplasm is the undifferentiated part of the cytoplasm
which surrounds and separates the neurofibrillae. It is also called inter fibrillar substance.

The chromophil substance also called as Nissl bodies are characteristic of nerve
cell body. They found in the form of irregular clumps or granules in the nueroplasm between
the bundles of neurofibrils. They are basophilic. They are seen in the dendrites also but not
in axons and axon hillocks and larger in motor neurons. E.M. studies reveal that Nissl bodies
are composed of dense network of rough surfaced endoplasmic r etriculum with RNA granule
on the outer surface of the double membranes. Their presence indicates high metabolic
activity in synthesizing neuro transmitter proteins, essential for replenishment of cytoplasmic
material in the axon. Following repeated stimulation, during degeneration and regeneration
after an injury to an axon or in other pathological conditions, the Nissl material undergoes an
apparent reduction or disappearance (chromatolysis).

Nerve cells may also show some inclusions or pigments under cer tain conditions.
Two kinds of pigments are seen lipofuchsin and melanin. Lipofuchsin is yellowish or brown
insoluble in usual fat solvents and appear in the form of granules dispersed throughout the
cell. This pigment is not present in this newborn but appears in increasing amounts with
advancing age.

Melanin appears in the form of dark brown granules in nerve cells of the olfactory
bulb, in the floor of IV ventricle, in the substantia nigra of midbrain etc. Its significance is not
known.

Dendrites appear as unmodified extensions of cell body. These branch repeatedly

and terminate near the cell body to produce terminal arborization (telodentria). They contain
neurofibrillae, nissl bodies and neuroplasm, like the cell body.

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 HISTOLOGY LECTURE NOTES

Fig.3.20 A multipolar neuron

1. Cell body
2. Dendrites
3. Nissl substance
4. Nucleus
5. Axon hillock
6. Axon
7. Myelin sheath
8. Node of Ranvier
9. Sheath of Schwann cell
10. Nucleus of Schwann cell
11. Motor end plates

Axon or Axis cylinder: The single axon arises from a special part of the periphery of
cell body called axon hillock. This area is devoid of nissl bodies and Golgi apparatus. Length
of axon varies from a fraction of a millimeter to several feet. Branches arise at right angles
from axons and are called collaterals. Axon is made up of fibrillae embedded in axioplasm
and covered by a membrane axolemma.

Nerves fibres are formed by the axon and its sheaths. All peripheral nerve fibres
consist of the axon covered with a thin protoplasmic sheath called neurolemma or sheath of
Schwann. Another sheath myelin or medullary sheath seen in some nerve fibres is next to
the axis cylinder and nuerolemma. These latter fibres showing myelin sheath are called

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 HISTOLOGY LECTURE NOTES

myelinated or medullated nerve fibres. Those without myelin sheath are amyelinated or non
medullated nerve fibres.

Neurolemma or Sheath of Schwann: This is a thin tube like membranes or sheath,

containing a nucleus surrounded by cytoplasm and is also known as Schwann’s cell.
The neurolemma forms sheath of the axon in all peripheral nerves and inside the central
nervous system there is no neurolemma but the nerve fibres have a neurolemma and axon
known as myelin or medually sheath. In those nerve fibres where there is no myelin sheath ,
neurolemma closely invests the axon.

Medullated or Myelinated nerve fibres: The medullated nerve fibres of the

peripheral nervous system consist of the axon, myelin or medullary sheath and neurolemma.
In the central nervous system, instead of the neurolemma, there is an irregular covering
made of glial cells.

Myelin forming the myelin sheath is glistening while when fresh, but with osmic acid
fixation becomes grey or black. In ordinary preparations, the myelin is dissolved by fat
solvents and in its place a clear space is left. This space is bridged by fine trabeculae-
neurokeratin network. Myelin sheath is not continuous but interrupted at intervals of 80 to
600 microns by constrictions called the nodes of Ranvier. Myelin sheath is absent where the
fibre branches and also at the beginning and at termination of the fibre. In certain
preparations, the myelin between the node of Ranvier is broken up by oblique clefts or
fissures extending from the surface called clefts of Schemidt-Lantermann celft and the
segments between them are the Schemidt Lantermann segments. At the nodes of Ranvier
the neurolemma sheath dips in and come in contact with the axon.

Amyelinated or non-myelinated nerve fibres (fibres of Remak): There is no

medullary or myelin sheath Axis cylinder is very slender and the thin neurolemma sheath
contains scattered nuclei of Schewann cells. There are no nodes of Ranvier, up to a dozen
or more of these nerve fibres are enclosed by the cytoplasm of the same sheath of Schwann
cells. Majority of the axons of autonamic ganglia and the processes of many of the smaller
cerebrospinal ganglia have no myelin sheath. In the CNS there are many non -myelinated
nerve fibres (without neurolemma).

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HISTOLOGY LECTURE NOTES

Fig.3.21 Neurolemma
1. Neurofibril
2. Myelin sheath
3. Nucleus of Schwann cell
4. Neurilemma of Schwann
cell
5. Axon
6. Node of Ranvier
7. Axolemma

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 HISTOLOGY LECTURE NOTES

Fig.3.22Stages in
formation of Myelin
sheath by a
Schwann cell
1. Schwann cell
2. Axon
3. Nucleus
4. Cytoplasm
5. Axolemma
6. Neurofibril
7. Neurilemma
8. Myelin sheath

Types of Neuron:

1. Sensory neurons convey impulses from receptors to the CNS. Processes of these
neurons are included in somatic afferent and visceral afferent nerve fibers. Somatic afferent
fibers convey sensations of pain, temperature, touch, and pressure from the body surface. In
addition, these fibers convey pain and proprioception (nonconscious sensation) from organs
within the body (e.g., muscles, tendons, and joints) to provide the brain with information
related to the orientation of the body and limbs. Visceral afferent fibers transmit pain
impulses and other sensations from internal organs, mucous membranes, glands, and blood
vessels.
2. Motor neurons convey impulses from the CNS or ganglia to effector cells.
Processes of these neurons are included in somatic efferent and visceral efferent nerve
fibers. Somatic efferent neurons send voluntary impulses to skeletal muscles. Visceral
efferent neurons transmit involuntary impulses to smooth muscle, cardiac conducting cells
(Purkinje fibers), and glands.

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 HISTOLOGY LECTURE NOTES

3. Interneurons, also called intercalated neurons, form a communicating and

integrating network between the sensory and motor neurons. It is estimated that more than
99.9% of all neurons belong to this integrating network

Fig.3.23 Types of neurons

A – Purkinje cell
B – Short neuron
C – Spinal ganglion cell
D – Pyramidal cell
E – Motor neuron
F- Bipolar neuron

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 HISTOLOGY LECTURE NOTES

Blood supply of nerve fibres: Nerves are profusely supplied by blood vessels.
There are inter-fascicular, perineurial, intra-fascicular arteries and arterioles. Endoneurium
contains a capillary network.

Physiological properties of the nerve fibre: The nerve fibre is essentially a highly
irritable conductor. Along it the dynamic nervous excitation propagates in waves, faster in
large than in small axons. During the conduction of excitation the activity of one portion of
the axon, serves as a stimulate activating the next portion, and so on. As the nerve fibres
become active it changes in its electric potential, the outside of each active portion becoming
negative relative to resting portion. Muscle action currents the flow of impulse between active
and resting regions. Conduction of nervous impulse is influenced by the permeability of axon
membrane, permitting sodium ions to enter the axon and potassium ions to migrate from it.
Nerve fibres can be classified into three types A, B and C according to the speed of
conduction. The ‘A’ fibres, with high velocity of conduction have a la rge diameter, a thick
myelin sheath and long internodes. The ‘B’ fibres are small myelinated fibres with relatively
short internodes. The ‘C’ fibres with the lowest conduction rate, have little or on myelin.

Nerve Terminations

Axons which form peripheral fibres terminates in peripheral structure to which or from
which they convey nerve impulses. Efferent fibres terminate in tissues which are excited
into activity by the nerve impulses; somatic efferent fibres terminates in voluntary, striated
muscle and visceral efferent fibres terminates in smooth muscle or secretory epithelium.
These are called effector organs. Afferent fibres end freely in tissues or in specially
organized structures. In either case they receive stimuli which cause them to carry nervous
impulses to the CNS. These nerve endings are called receptors.

Effectors:

Motor end plate (somatic efferent fibres): Nerve fibre enter the perimysium and
branch to pass to individual muscle fibres where they end in structures called motor end
plate. At the end plate region the myelin is lost and endoneurium becomes continuous with
endomysium of muscle fibre. The axon devoid of its sheath divides to fine terminations
which are in close contact with the sarcolemma. The nerve ending does not penetrate into
sarcolemma. Beneath the end plate region, there is an increase in the number of muscle
nuclei and this region is known as the sole of motor end plate.

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 HISTOLOGY LECTURE NOTES

Visceral efferent fibres: These are myelinated fibres terminating in cardiac muscle,
smooth muscle of viscera, blood vessels, hairs and glands. The nerve fibres form plexuses
around the muscle bundles. From these plexuses fine fibres pass to individual muscle fibre.
In glandular epithelium, the nerve fibres from plexus beneath the basement membrane
through which fibres pass to end on gland cells.

Receptors: are of many kinds those concerned with general bodily sensibility or
somaesthetic sensibility touch, pressure, heat, cold, pain, temperature, position, movement),
and those which form organs of special senses (smell, sight, taste, hearing and head position
and movement). They may also be classified as exteroceptors the receptors, affected by
external stimuli (touch, light, pressure, cutaneous pain and temperature, smell, sight and
hearing) proprioceptors those affected by stimuli arising within the body wall, especially those
of movement and posture; visceroceptors those affected by stimuli arising within the Viscera.
All receptors, depending upon their terminal arborizations, come under two classes: F ree or
encapsulated.

Free or Nonencapsulared endings: These are found practically in all epithelia,

connective tissue, in muscle and serous membranes. Here the fibres after losing their myelin
sheath form a subepithelial plexus, from which the axons send their branches between cells
and end in knob like terminal swellings.

Encapsulated endings: They include end-bulbs, Meissner’s corpuscles, pacinian

corpuscles, muscle spindles and muscle tendon spindles. These are all characterized by a
capsule of more or less flattened connective tissue cells, which enclose the terminations of
the axon.

Ends bulbs (Krause’s end bulbs): Spherical or oval in shape, consists of a thick
lamellated capsule of connective tissue cells and fibres surrounding a central cavity or i nner
bulb. Within this end bulb, the naked axons of one or more myelinated fibres end either
simply or in the form of skeins. End bulbs are found in lips, tongue, cheeks, soft palate,
epiglottis, nasal cavities, lower end of rectum, peritoneum and other serous membranes
tendons, ligaments, connective tissue of nerve trunks, synovial membranes of certain joints,
glands penis and clitoris. These are receptors for cold.

Meissner’s corpuscles or tactile corpuscles: Occurs in hairless portion of skin and

especially most numerous in finger tips, palm of hand and sole of feet. They lie within the

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connective tissue of dermal papillae. They are oval in shape and composed of flattened,
horizontally arranged connective tissue cells and lamellae surrounded by connecti ve tissue
capsule. One myelinated fibre is distributed to each corpuscle. After losing the myelin the
naked axons pass into the corpuscle branch, and pursue a spiral course among the
connective tissue elements. These corpuscles are concerned with the sen se of discriminative
touch.

Pacinian corpuscles: These are lamellated elliptical structures. They are large and
visible to naked eye. Each corpuscle is formed of a large number of concentric lamellae of
connective tissue fibres, lined by a single layer of connective tissue cells. The spaces
between the lamellae filled with the fluid or semi-fluid substance. There is an inner bulb in
the centre. Each corpuscle is supplied with a single myelinated naked axon extends through
the centre of the inner bulb and ends in a knob like expansion. Fine capillary networks are
seen within the lamellae but they do not enter the inner bulb, pacinian corpuscles are found
in deeper subcutaneous connective tissue of hand and foot, peritoneum, pancreas,
mesentery, penis, clitoris, urethra, nipple, mammary gland, connective tissue in the vicinity
of tendons, ligaments, joints. They are stimulated by deep or heavy pressure.

Muscle spindles (Neuromuscular spindles): In voluntary muscle, sensory nerves

end in end bulbs and muscle spindles. The muscle spindles, is a cylindrical elongated
structure within which are one or several muscle fibres, connective tissue blood vessels and
myelinated nerve fibres. The whole is enclosed in a connective tissue capsule, which is
pierced, at various points by one or more nerve fibres. The thicker myelinated sensory fibres
lose their myelin, as they branch repeatedly within the spindle. The axons terminate in close
opposition to the sarcolemma of the enclosed muscle fibres. The muscle fibres of the muscle
spindle are thinner, richer in sarcoplasm and have more nuclei.

Muscle tendon spindle (Organ of Golgi): Found at the junction of muscle and
tendon, spindle-shaped, composed of tendon bundles covered by a connective tissue
capsule. Into this enter one or several afferent nerve fibres which terminate in arborization
around bundles.

Muscle spindles and muscle tendon spindles are proprioceptors of position and
movement receiving stimuli due to constant or varying tensions of voluntary muscle and their
attached tendons.

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Fig.3.24 Encapsulated endings

The Synapse: The relations between two neurons are of the nature of contact rather
than complete structural continuity. This junction of two neurons is called a synapse.
The synapse may be defined as a point of contact of the cell membrane (plasma membrane)
of an axon terminal with the cell membrane of another neuron. The synapse may be between
the axon termination of one neuron and the cell body of another (axo -somatic synapse) or it
may be between the axon terminals and the dendrites of another neuron (axo-dendritic

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 HISTOLOGY LECTURE NOTES

synapse). In the CNS, interwoven telodendria of Axon and dentrite arborization forms a
dense feltwork of fibres which termed neuropil.

Fig.3.25 Muscle tendon

1. Axon
2. Sarcolemma
3. Sarcoplasm
4. Subneural cleft of motor
end plate
5. Mitochondrion in axon
6. Mitochondrion in muscle
7. Synaptic vesicle

The axon terminals of a single axon may from synapses with both the body and
dendrites of another neuron. Various methods of contact occur among which are (1) by
neuropodia or terminal buttons (2) by gemmules, (3) by parallel opposition. In the first
method the axon terminals end in small bulb like expansions or neuropodium consists of
neurofibrillae. Loops embedded in perifibrillar substance, some times there are simply small
neurofibrillar rings. In the gemmule type of synapsis axon processes running at right angles
to the dendrites of another neuron appear to come in contact with their gemmules. In the
third type the axon terminal comes into length wise opposition with the dendrite cell body or
in some cases, the amyelinated portion of the axon of the receiving neuron. It is obvious that
one axon may carry impulses to a number of neurons and that each neuron may receive
impulses from the axons of a number of other neurons.

GANGILIA

Ganglia consist of aggregations of neurons situated outside the central nervous

system. Each ganglion is surrounded by a connective tissue capsule which is continuous
with the epineurium and perineurium of the peripheral nerve. Connective tissue trabeculae
from the capsule extend into the ganglion to form a framework. Within the ganglion the
nerve cells are separated into irregular groups by strands of connective tissue and by
bundles of nerve fibres.

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 HISTOLOGY LECTURE NOTES

Each ganglion cell is enveloped by a double layer of cells. The outer part of capsule
is composed of flat fibroblasts and connective tissue fibre which are continuous with the
endoneurium of the associated nerve fibre. The inner part consists of a layer fusiform of
satellite cells or amphicytes, which derived from the same source as the ganglion cells
themselves.

Cranial and spinal ganglia: They are made up of pseudo-unipolar or ‘T’ bipolar
neurons with prominent nucleolus. Chromophil substance may either be scattered
throughout the cytoplasm as diffuse granules or it may form rather small finely granular
substance. Nissl bodies which are concentrically arranged around the nucleus. Most of
these cells have one Principal myelinated process which at some distance from the cell
body, divides into a peripheral nerve and a central branch which enters the CNS as a
sensory root fibre.

Fig.3.26 Ganglion cell

1. Cell
2. Amphicytes

Autonomic ganglia: The majority of the sympathetic and parasympathetic ganglia

have a connective tissue framework and multipolar neurons with numerous nerve fibre.
The cell bodies vary in size from 20 to 45 microns. The nucleolus is relatively large and
vesicular round or oval and has one or more well defined nucleoli. The nucleus is often
eccentrically situated. Binucleated neurons are not uncommon and multi nucliated neurons
have also been seen. Chromophil substance or Nissl bodies may be distributed uniformly
throughout the cytoplasm or may be confined to the perinuclear zone or to peripheral
cytoplasm. In large ganglia, stellate cells are present. The dendrites either terminate within
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 HISTOLOGY LECTURE NOTES

the capsule or pass through capsule to form intercellular plexus of fibres depending on their
length. Often two ganglion cells share a single capsule in smaller terminal ganglia such as
those in the intestinal wall, many of the ganglion cells lack capsule parasympathetic ganglion
cells are found in almost all visceral structures.

NEUROGLIA

This is the interstitial tissue of the nervous system. In the brain and spinal cord the
connective tissue is limited to the enveloping membranes the meninges and a slight amount
accompanies the blood vessels. The supportive framework the interstitial tissue is formed by
a special group of cells and fibres called neuroglia.

In ordinary preparations of CNS many nuclei are seen which belong neither to nerve
cell nor vascular tissue. These nuclei which do not possess nucleoli belong to Neuroglia
cells. The neuroglia (often simply termed glia) may be divided into the following classes.
(1) Astroglia or Astrocytes (2) Oligodendrgolia (3) Microglia or mesoglia and (4) Ependyma.
Except microglia all of the above are of ectoderm origin.

Special and often difficult methods should be employed to study details neuroglia.
What are described below will not be seen in ordinary preparations.

1. Astrocytes: These are stellate cells with many processes. They are of two
varieties protoplasmic astrocytes of numerous processes which extend from the cell body in
all directions and branch frequently. The nucleus is oval and has scan ty chromatin and has
no nucleolus. Small granules present in the cytoplasm of cell body and processes are called
Gliosomes. They are found principally in the grayt matter of brain and spinal cord. They
partly envelop the neuron bodies to form satellite cells. One or more of the processes of
these astrocytes have peculiar terminal expansions called foot plates or perivascular feet
which are applied walls of capillaries or small blood vessels. The fibrous astrocytes differ
from the protoplasmic astrocytes having fewer processes. But these are most straight and
longer. They possess gliosomes and coursing through their cytoplasm are fit straight,
unbranched fibres the neuroglia fibres. The fibrous astrocytes have perivascular feet and are
found chiefly in the white matter.

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 HISTOLOGY LECTURE NOTES

Fig.3.27 Astrocytes
1. Nerve cell
2. Foot plate
3. Blood vessel

2. Oligodendroglia: These are smaller than astrocytes. Their processes are few
smaller and more delicate. They have no perivascular feet but th eir cell body may be applied
to surface of a capillary as perivascular satellite. They are present both in grey and white
matter. In grey matter they form perinueuronal satellite to the neuron bodies. In white matter
they lie in row between myelinated nerve fibres to form an investment around the myelin.
They are known as inter-fascicular glia. Their relation to myelin sheath in CNS is similar to
that of Schwann cells in PNS.

3. Microglia: These are small cells with deeply staining small nuclei often elongated
or irregular shape they resemble fibroblast nuclei. They have neither gliosomes nor fibres.
They are found both in the grey and white matter but a less in numbers in the latter. In the
grey matter, they form perineuronal satellite and although they dont have perivascular feet
they form perivascular satellites.

4. Ependyma: In ordinary preparations appears to consist of closely packed

elongated nuclei lining the central canal of spinal cord and ventricles of brain. Their long axis
is perpendicular to the cavity and they present the appearance of columnar epithelium. Their
process ramifies more or less deeply in the wall of the neural tube. They show neuroglia
fibres and hence they are regarded as neuroglial cell. They may have in certain places, cilia
which protrude into the neural cavity.
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 HISTOLOGY LECTURE NOTES

Fig. 3.28 Neuroglia

A. Oligodendroglia
B. Microglia
1. Nucleus
2. Cell body
3. Process

Fig.3.29 Ependyma
1. Epithelial cell
2. Cilia
3. Process

Repair and Regeneration of Adult tissues:

(a) Epithelium: In contrast to the more highly differentiated tissues like the nerve
cells and skeletal muscle fibres epithelium has a capacity for cell division and repair if the
injury is not very extensive.

(b) Connective tissue proper has the greatest powers for regenerations and repair
and is the tissue involved in the healing process of wounds and the regenerating tissue is
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 HISTOLOGY LECTURE NOTES

known as granulation tissue is known as granulation tissue. Regeneration of cartilage is a

slow process and possibly does not occur at all in many instances. Osseous tissue has
powers of regeneration and repair.

(c) Smooth muscle has limited powers of regeneration that of skeletal muscle is
restricted, while the regenerative capacity of adult heart muscle is very slight.

(d) In nervous tissue in the adult, when nerve cell body dies, there will be no
regeneration or replacement by mitotic division of existing cells. But when the cell body is
intact the nerve fibre or axon alone is injured distal to the injury there is degeneration leading
to complete break down and disappearance of the portion of the nerve fibre including its
terminal arborization (Wallerian or secondary degeneration). But proximal to the inj ury, after
certain initial degenerative processes, regeneration occurs leading to a new growth from the
injured end. Changes in the nerve cell body (e.g) Chromatolysis also occur and fibre
regeneration occurs only if the cell body survives, it depends also on the type of neuron,
nature and location of the site of injury etc.

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SPECIAL HISTOLOGY

4. CIRCULATORY SYSTEM

The circulatory apparatus consists of a cardio vascular system and lymphatic system.
Cardiovascular system consists of (a) a central propelling organ- the heart (b) a series of
efferent tubes - the arteries, which by branching constantly increase in number and
decrease in caliber and which serve to carry the blood form heart to the tissues, (c) minute
anastamosing tubules - the capillaries into which the arteries empty and through the walls of
which the interchange of elements between blood and the other tissue takes place, (d) a
system of converging tubes (the efferent tubes) - the veins which receive blood form the
capillaries, unite to form large vessels as they approach the heart; and serve for draining the
blood from that organ.

The lymphatic system consists of lymphatic glands or nodes and lymphatic

capillaries and various sized lymphatic vessels, which ultimately drain into two main trunks,
the thoracic duct and right lymphatic duct.

BLOOD VASCULAR SYSTEM

The entire system the heart, arteries, veins, capillaries has a common lining, which
consists of a single layer of cells forming the endothelium. In the capillaries this single layer
of cells forms the entire wall. In the heart, arteries and veins the endothelium is invested with
accessory coat of muscle and connective tissue.

Capillaries:

These are delicate tubes whose average diameter is about 7 to 9 microns. Capillaries
branch extensively without change in caliber and these branches anastomose to form
extensive networks whose meshes vary in size and shape in different tissues and organs.
The greater the metabolic activity of the tissue, the denser is the capillary network.

The wall of a capillary consists of a single layer of flattened endothelial cells (simple
squamous epithelium) separated by narrow intercellular spaces, which are filled with a little
intercellular cement. On surface view the cells appear as a delicate mosa ic, which can be
demonstrated by precipitation of silver in the intercellular clefts. The cell borders are usually
serrated or wavy. Cells are arranged with their long axes parallel with the long axis of the

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 HISTOLOGY LECTURE NOTES

tube. In capillaries of small caliber the cells are very narrow and their ends pointed. In wider
capillaries cell are oval with blunt ends. Cytoplasm is clear or finely granular and nucleus is
central and elongated. Cells are thicker in the region of the nucleus. In fixed preparation,
there is pronounced bulging of nuclei into the lumen, due to shrinking of cytoplasm.
The nucleus is flattened and surrounded by a small amount of cytoplasm and is usually
found pressed against the cell wall. Even one is sufficient to form the circumference of a
small capillary. For the larger ones, 3 to 5 cells are required.

The part of a capillary, which is nearest to the arteriole, is called an arterial capillary
and that part nearest the venule is called a venous capillary but these designations are
based on topography and function.

The sinusoids or sinusoidal capillaries are irregular blood filling spaces viz., liver and
other organs. These are larger, abou 25-30 microns in diameter. Lining cells have processes,
which extend into the lumen of the sinusoids. The connective tissue which usually surrounds
capillaries is so greatly reduced in amount in sinusoids that the lining cells of the latter are
closely applied to the parenchyma of the organs, being separated form the latter by a fine
network of reticular fibres. Lining cells of sinusoids are either phagocytic or non-phagocytic
cells. The former belonging to the reticulo-endothelial system.

The capillary walls furnish the membrane for diffusion, filtration and osmosis of fluids
to and from the blood stream. The passage of fluid through the walls of the capillary is
partially dependent on the blood pressure within the capillaries and on the colloid osmotic
pressure of the blood which play a significant role in increasing capillary permeability.

ARTERIES

The wall of an artery is composed of three tunics or coats which are (1) innermost
coat - the Tunica intima consisting of endothelium continuous with that of capillaries, (2) the
middle coat - Tunica media consisting mainly of smooth muscle with varying amounts of
elastic and collagenous fibre and (3) the outer coat - Tunica adventitia or externa
composed of loose connective tissue. The structure of these coats varies according to the
artery and it is convenient to distinguish (a) small arteries or arterioles, (b) medium-sized
arteries and (c) large elastic arteries.

Small Arteries (Arterioles)

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 HISTOLOGY LECTURE NOTES

The three coats are distinguishable as an endothelial intima, a muscular media and
an adventitia of connective tissue. An internal elastic membrane is present and seen e asily in
arterioles of 40 microns diameter and upward. This marks the boundary between the tunica
intima and media. The tunica media is made up of smooth muscle fibres circularly arranged
formed of one or two layers only in arterioles and gradually increasing in thickness, as the
caliber of the vessel increases. The Tunica externa is made up of flattened fibroblasts and
longitudinally arranged collagenous fibres and as the caliber of the vessel increases elastic
and reticular fibres are also present. These vessels serve more for distribution of blood to
various capillary beds and to control blood pressure.

Medium sized (Muscular) arteries

These include all named arteries excepting very large ones. The walls of these
vessels are very thick due to large amount of muscular tissue in the media. These are called
muscular arteries. They are also called ‘distributing’ arteries because they distribute blood
to all organs and regulate the supply according to functional needs. These Tunica int ima
exhibits three definite layers endothelium, intermediate layer and internal elastic membrane.
The intermediate layer consists of delicate elastic and collagenous fibrils and a few
fibroblasts.

The internal elastic membrane is a thick fenestrated band. It marks the boundary
between the intima and media owing to the large amount of muscle in their walls. The
postmortem contraction of arteries throws the elastic membrane into longitudinal folds hence
in transverse sections it has the appearance of a corrugated or wavy band. The med ia is the
thickest coat and has layers of circularly disposed muscle fibres. There circularly disposed
elastic nets in the media.

The adventitia consists of a thick layer of connective tissue containing collagenous

and elastic fibres disposed off longitudinally. Between the adventitia and the media is a very
well defined external elastic membrane. The outer layer of adventitia blends gradually with
the surrounding connective tissue when attaches the artery to other structures.

Larger arteries (Elastic arteries)

Also called conducting arteries because they conduct blood from the heart to the
muscular arteries. The wall is relatively thin for the size of the vessels. The chief
representative of this type is the aorta. The T. intima is made up of endothelium and
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 HISTOLOGY LECTURE NOTES

endothelial cells are short and polygonal. The deeper portion of intima is made up of coarse
collagenous fibres; some longitudinally oriented smooth muscle fibres and longitudinally
arranged elastic fibres. The elastic membrane is usually split into two or more lamellae,
which merge with other similar membranes, both in the media, and hence it is difficult to
identify it. The muscle tissue is relatively reduced in amount. The adventitia is very thin and is
composed of connective tissue mostly of collagenous fibres arranged in longitudinal spirals.

VEINS:

The caliber of veins is as a rule much larger but the walls are thinner than arteries
due to a great reduction of the muscular and elastic elements. The collagenous connective
tissue is present in much larger amount and constitutes the bulk of the wall. The three coats
are present but their boundaries are often indistinct. The whole wall is flabbier and tends to
collapse when not filled with blood.

The histological structure of the veins varies greatly and the variations are not always
related to the size of the vessels but depend on local mechanical conditions. The structure
may be quite different in veins of same caliber and even in different portions of the same
vein.

Small veins or venules:

These are endothelial tubes. Tunica intima surrounded by collagenous fibres and
fibroblasts, with a few isolated, circularly disposed, plain muscle fibres forming the Tunica
media. As the caliber of the vessel increases, circular smooth muscle fibres forms a
continuous layer and may also become many layered, the individual layers being separated
by loose connective tissue. The Tunica adventitia is relatively thick and is made up mostly of
longitudinally disposed collagen fibres.

Medium sized veins

These include all the named veins and their immediate branches excepting the main
trunks. The Tunica media is composed of circularly arranged plain muscle fibres and
collagen fibres. The media is thickest in the veins of the limbs and is very thin in the views of
the head and abdomen. The Tunica adventitia is well developed and forms bulk of the wall. It
consists of collagenous and elastic tissues.

Large veins
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 HISTOLOGY LECTURE NOTES

The Tunica intima is made up of endothelium. The Tunica media is usually thin with
the reduction in circularly arranged smooth muscle fibres. Tunica adventitia is very thick and
among collagenous and elastic fibres, there is dense arrangement of longitudinal bundles of
plain muscle.

Valves: Veins over 2 m.m. in diameter are provided with valves at intervals. These
are semilunar flaps, which project into the lumen, their free margin being projected towards
the heart. These are derived from the intima and consist of loose connective tissue covered
by a single layer of endothelium.

LYMPH VASCULAR SYSTEM

Besides the blood vessels, the body contains a collateral system of channels, lined by
endothelium which collect the tissue fluid and return into the blood stream. The fluid in these
vessels is lymph. Unlike blood, the lymph circulates in one direction only, from the peripher y
towards the heart. The lymphatic capillaries starts blindly from the tissue collect the lymph
and freely anastomose along their course and lymph passes through one or more lymph
nodes in the course of lymph vessels before entering into the blood stream.

Lymph capillaries: Are like blood capillaries but are larger. Lymphatic capillaries are
present in most tissues and organs except the central nervous system, bone narrow, eyeball,
internal ear and foetal placenta.

Lymph vessels: These are like veins in structure but their walls are thinner than
veins of corresponding caliber. They contain numerous valves, which are directed in the
direction of lymph flow.

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5. GLANDS

Certain cells of the body in addition to carrying out some metabolic processes
necessary for their own existence, also manufacture specific substance not for their own use,
but to be extruded, from the cells and used elsewhere in the body – secretion (e.g. gastric
juice) or discarded - excretion (e.g. urine). Such cells are known as gland cells or glandular
epithelium. These cells are usually grouped into small units, enclosed and supported by a
connective tissue framework.

A gland may consist of a single cell, as for example, the goblet cell or the unicellular
glands of invertebrates. Most glands of the body are composed of more than one cell
(multicellular gland).

Classification of glands:

The glands of the body are classified as exocrine or endocrine. Exocrine glands have
ducts which carry out the secretion to an epithelial surface, while endocrine glands are
ductless and pour their secretion into the blood or lymph.

Exocrine glands

The exocrine gland consists of a secretory portion and a duct. The secretory portion
is arranged as a cluster of secretary cells known as a secretory unit or acini, which has a
central lumen, which opens into the first portion of the ductular system. The exocrine glands
can be classified in several different ways based on the shape and form of the secretory unit,
nature of secretion and mechanism of secretion.

Based on the shape of the secretory unit:

If the secretory units are tubular in shape it is a tubular gland. If secretory units are
more rounded, then they are called alveolar or acinar glands. Glands which have
characteristics of both these, or which have tubular portions with terminal alveoli, are called
tubuloalveolar glands.

Based on the ductular system:

Glands are classified into Simple and compound glands in this view. The ductular
system of a gland may consist of simple epithelial tube draining a secretory unit and opening

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at the epithelial surface or it may be in the form of an extensively branched system, the initial
smallest ducts draining the secretory units, uniting to form larger ducts at various levels.
Finally forming the main duct, which conveys the secretion from the gland to an epithelial
surface. Based on the ductular system glands are classified as simple glands, which has an
unbranched duct system, and compound glands which has a branched duct system.

A. Simple glands:

(1) Simple tubular glands: The glands consist of simple tubular secretory unit which
opens on the surface. All the cells may be secreting or only the deeply situated cells alone. In
the later case the more superficial portion of the tubule serves merely as a duct. The more
highly developed simple tubular glands consist of a mouth opening upon a surface, a neck
usually somewhat constricted and a fundus or deep secretory portion of the gland.

Fig.5.1 Epithelial glands

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 HISTOLOGY LECTURE NOTES

Simple tubular glands are divided according to the structure of the fundus into
(a) straight, (b) coiled and (c) branched tubular glands.

(a) A Straight tubular gland is one in which the entire tubule runs in a straight
unbranched course. e.g. glands of the large intestine. (b) A coiled tubular gland is one in
which the deeper portion of the tubule is coiled or convoluted. The sweat glands of the skin
are the most typical examples. (c) A branched tubular gland is a simple tubular gland in
which the deeper portion of the tubule branches, the several branches being lined with
secreting cells and opening into a superficial portion which serves as a duct e.g. fundic,
pyloric and cardiac glands of the gastric mucous membrane.

(2) Simple alveolar glands: These are found in branched form, e.g., Sebaceous and
Meibomien glands.

(3) Simple tubulo-alveolar or tubulo–acinar glands: These are found only in the
branched form e.g. Brunner’s glands of the duodenum.

B. Compound glands

These contain a large number of branching duct systems, which finally join to form a
common or main duct, which coveys the secretion from the glands. Based on the nature of
secreting units compound glands are classified into:

(1) Compound tubular glands: The terminal ends of the duct systems end in tubular
secretary units, which are usually tortuous or coiled e.g. kidney, Testis.

(2) Compound alveolar glands: The terminal part of the duct system ends in alveoli
with dilated sac-like lumen e.g. mammary gland.

(3) Compound tubulo-alveolar or tubulo-acinar glands: In these the secretary

units are with terminal or lateral alveoli. E.g. parotid gland, pancreas etc.

Large amount of connective tissue present in a compound gland, which starting off
from the capsule divides the gland into lobes by interlobar connective tissue septa. Each
lobe is subdivided by connective tissues into lobules, which are usually microscopical in size
called interlobular connective tissue septa. Each lobule is made up of a number of secretin g
units or acini, with the ducts and inter acinar connective tissue.

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Parenchyma- The secreting units may be tubular, alveolar or tubulo alveolar and are
made of secretory cells. The acinar duct is the smallest duct continuous with the lumen of the
acinus and is usually lined by flattened or cuboidal epithelium. The acinar ducts unite within
the lobule to form intralobular ducts and several of this type unites to form an Interlobular
duct, located in the connective tissue between the lobules. Interlobular an d interlobar ducts
are similarly formed, the latter uniting to form the main duct of the gland. Interlobular ducts
are lined by columnar epithelium but have a thin connective tissue coat. As the duct
increases in caliber the connective tissue coat also proportionately increases and the
epithelium may become two layered columnar.

Based on the mode of secretion:

This classification is based on the manner in which the secretory cells of the glands
elaborate their secretion. Based on this glands are classified into Holocrine, merocrine and
apocrine gland. In holocrine glands, the secretory products are accumulated in the
cytoplasm of a cell, which disintegrates fully and constitutes the secretion secreted e.g.
sebaceous gland. Merocrine glands elaborate secretion without losing any part of the cell
e.g. salivary glands. In Apocrine glands, a little or the cytoplasm along the free secretory or
apical borders of the cells is lost e.g. mammary gland.

Fig.5.2 Gland types based on

their mode of discharge

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Based on the nature of secretion:

Glands may be classified as cytogenous and non-cellular. Cytogenous gland

produces living cells as their secretion. Eg. Testis, Ovary. Non-cellular glands are further
classified into serous, mucous and mixed glands based on the consistency of their secretion.

Based on the consistency of the secretion:

Based on the consistency of the secretion glands are classified into Serous, mucous
and Mixed glands. Serous glands produce a whey-like serous secretion. Mucous gland
produces a slightly more viscid fluid. The glands that secrete mucin, glycoprotein which when
mixed with water become mucous are termed as mucous glands. Any gland that produces a
mixture of serous and mucous secretions is called a mixed gland.

Serous secretory unit: The nucleus of a serous secretory cell is usually rounded
and lies towards the base of the cell. At the base of the cytoplasm contains secretory
granules called zymogen granules. Cytoplasm and Zymogen granules stain with eosin
(acidophilic).

Mucous Secretory unit: The nuclei of the mucous secretory cells are flattened and
located at the base of cells that contain them. There is less chromoideal substance. Above
the level of the nucleus the cytoplasm contains mucin droplets, which impart a vacuolated
appearance to the very light staining cytoplasm, since with ordinary staining techniques
mucin is dissolved. Cytoplasm stains only lightly with haematoxylin. Mixed glands posses
both mucous and serous secretory units.

Endocrine glands

Certain glands are lacking in ducts and are termed endocrine glands. The endocrine
glands secrete specific chemical substance called hormones which have specific effects on
the other tissue or organs of the body e.g., thyroid, parathyroid, pituitary etc. The secretions
of these glands are conveyed directly into the blood stream. To facilita te this endocrine
glands have numerous sinusoids forming an extensive network, surrounding the secretory
cells, which are usually arranged in small groups, follicles or anatomizing cords of cells.

☺☺☺☺☺

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6. DIGESTIVE SYSTEM

The digestive system consists of the alimentary tract and other associated structures
like the tongue, teeth, salivary gland, liver and pancreas.

SALIVARY GLANDS

The Salivary glands may be classified on the basis of their secretion and the
epithelium of their end-pieces as serous, mucous and mixed glands. Of the three pairs of
salivary glands in mammals, the parotid glands is the serous type, the mandibular and
sublingual glands are of the mixed type.

The salivary glands are true compound tubuloalveolar glands, consisting of large and
small lobules. The glands are enveloped by a connective tissue capsule, which sends
trabeculae into the interior of the organ to form interlobar, interlobular connective tissue
framework.

1. Serous gland

Pyramidal cells resting on a basement membrane and enclosing a narrow lumen line
the serous alveoli. The cell boundaries are indistinct. Secretory capillaries extend between
the cells and communicate with the narrow lumen of the alveolus (intercellular secretory
canaliculi). Continuation of these from the so-called intracellular secretory canaliculi, which
are infoldings of the cell membrane. The spherical nucleus lies in the proximal half of the
cells but not against the wall. The serous cells show inclusions of strongly refractive
secretory zymogen granules which vary in number and arrangement in the cytoplasm which
is generally acidophilic.

Between the secreting cells and the basement membrane is a layer of flat, star
shaped myoepithelial cells. These are joined to each other by their cytoplasmic processes
to form a basket like shell around the secretory cells. Therefore they are called basket cells.
These are believed to assist in the discharge of secretion into the ducts by their contraction.

Ducts: The alveoli open into narrow acinar ducts lined by cuboidal epithelium. In
salivary glands, the acinar ducts are also called as intercalated ducts. Within the lobule, the
acinar ducts unite to form Interalobular ducts, which are lined by simple columnar epi thelium,
with intensely eosionophilic cells, which show basal striations. These are believed to posses

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a secretory function (secretion of calcium salts) and these ducts are also referred as striated
tubules or salivary ducts. A thin connective tissue layer, containing collagen and elastic
fibres and few muscle cells, surrounds the simple columnar epithelium resting on a basement
membrane.

The interlobar ducts are also lined by simple columnar epithelium, but the cells do not
show striations. The larger ducts show a two layered columnar epithelium and the terminal
portions show stratified columnar epithelium, which changes to the stratified squamous type
at the openings of the ducts. There is gradual increase in the connective tissue layer as the
ducts become two layered. In certain instances, like parotid gland of ruminants, mast cells
are found around the salivary ducts.

Fig.6.1 Histology of salivary gland

1. Serous acinus (L.S)
2. Mucous acinus (T.S)
3. Serous demilune
4. Mucous acinus (L.S)
5. Inter-calated duct
6. Striated duct
7. Basket cell

2. Mucous gland

Pyramidal or polygonal cells resting on a basement membrane line the mucous

alveoli. The cell boundaries are distinct and the nucleus is flattened lies against the base of
the cell and stains deeply. Secretory capillaries are absent. With haematoxylin and eosin
staining, the cytoplasm of the mucous cells stains blue. When full of secretion, mucous cells
are often markedly distended and occupied, except for a narrow strip of basal cytoplasm by
mucinogen droplets, demonstrable only by special methods. They may be stained with mucin

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stains (e.g. mucicarmine). The pressure of the contents flattens mucous cell and nucleus is
pushed against the basal wall.

At the base of the mucous alveolus, a crescent shaped groups of serous cells,
referred to as the demilunes or crescents of gianuzzi. These may have direct access to
the lumen of the alveolus or may communicate with it by means of intercellular secretory
canaliculi, which pass between the mucous cells.

In addition to the true demilunes there are also groups of non -secreting mucous cells
called Stohr’s crescents. These lack secretory capillaries.

The mixed glands either have separate serous alveoli or the same alveolus may
contain both mucous and serous cells singly or in groups. In the second case, secretory unit
is usually elongated and the serous cells usually occupy the terminal blind end of the
alveolus, with mucous cells nearer the exit.

TONGUE

This organ consists essentially of a lining of stratified squamous epithelium and

striated muscle, arranged in a number of layers. The connective tissue attaches the tough
mucous membrane of the tongue to the muscular mass. The epithelium of the tongue is
thickest and has the heaviest stratum corneum on the dorsal surface. The tongue bears
various papillae, which are named for their characteristic gross morphology. The papillae are
macroscopic projections formed with a central core of connective tissue and a covering layer
of stratified squamous epithelium. The connective tissue core may give rise to small
microscopic papillae (commonly referred to as papillary bodies) over which the epithelium is
moulded. According to the shape, the (macroscopic) papillae of the tongue are divided into
various types.

(1) Filliform, (2) Fungiform (3) Circumvallate (present in all animals) and (4) Foliate
(present in horse, donkey, rabbit).

(1) The filliform papillae consist of a connective tissue core derived from the lamina
propria and an epithelial covering characterized by a heavy stratum corneum. The papillae
are usually slender and pointed. In most species the papillary core does not extend above
the level of the glossal epithelium. The visible projection is made up entirely of epithelium.

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Large conical papillae, whose core projects beyond the surface of the tongue, occur in all
domestic mammals except the horse and donkey.

(2) The fungiform papillae: These are relatively few in number and interspersed
among the filiform palpillae. They have rounder and broader summits and narrow attached
ends. The connective tissue core is rich in nerves and characterized by papillary body, have
relatively less cornified epithelium containing taste buds. Taste buds may or may not be
present.

(3) The circumvallate papillae: These are very few in number and are arranged in
’V’ shaped row nearer the posterior part of the tongue. They resemble fungiform papillae but
are much larger and are surrounded by a cleft (moat) lined with epithelium. They project
above the lingual epithelium only slightly or not at all. Their connective tissue core bears
microscopic papillae and is rich in nerves and lymphocytes. The epithelial surface facing the
moat contains many taste buds. Deeper to that papillae lie groups of serous glands called
Van Ebner’s glands whose excretory ducts open into the moat at various levels.

(4) The foliate papillae: These are present only in some animals and each consists
of a series of parallel connective tissue leaves, rich in nerves and bearing secondary papillae
that project into the covering stratified squamous epithelium. Gustatory furrows separate
them from each other. The epithelium covering the sides of the leaves bears taste buds.
Deeper to the papillae lie serous glands whose ducts empty into the gustatory furrows.

Fig.6.2 Papillae in the tongue

A. Filiform papilla
B. Fungiform papilla
C. Circumvallate papilla
D. Foliate papilla
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Taste buds:

These are microscopic structures found in the epithelium of the fungiform, foliate and
circumvallate papillae of the tongue; they are also found widely separated in the soft palate;
epiglottis and the free edge of the vocal folds. They are ellipsoid bodies embedded in an
upright position in the epithelium of the mucous membrane. The taste bud is made up of
supporting or sustentacular cells and neuroepithelial cells. The peripheral supporting
cells form the outer layer of the taste bud. These are curved narrow cells with an ellipsoid
nucleus, which surrounds the central supporting cells which are shorter and straighter than
the peripheral ones and rounded off on the distal end. In some species the taste buds also
contain basal supporting cells. These lie deep to the other supporting cells and are
connected to them by means of processes.

The neuroepithelial cells lie among the supporting cells. These are slender cells
thickened slightly in the region of the nucleus and resemble the central supporting cells.
Each is characterized externally by a fine hair-like process called taste hair which projects
through a minute opening in the epithelium called the taste pore. The inner end of the cells
tapers to a fine process, which may be single or branched. The sensory nerve fibre which
convey gustatory impulses end within the taste buds in a network of varicose fibres called
intragemmal fibres. The terminations of the intragemmal fibres end directly on the cells.
Some of the termination also end between the cells (inter gemmal fibres). Dissolved
substances stimulate the sensory cells.

The lingual glands are situated partly below the mucous membrane, partly in the
inter-muscular tissue, and are especially abundant on the root, on the margins and near the
circumvallate papillae. Those in the region of circumvallate papillae are the serous glands of
Van Ebner. The remaining glands are all mucous glands.

Species difference:

1. Filliform Papillae: In ruminants the connective tissue core gives rise to several
small secondary papillae, whereas the epithelial coat is raised into a single cornified one. In
the horse, donkey and pig, the papillary core is an enlarged similar to papilla, from the top of
which a cornified thread projects above the epithelial surface. In carnivores, the connective
tissue core extends above the surface epithelium and bear papillae of unequal sizes.

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2. Fungiform papillae: Taste buds are very few in the Ox and horse. They are
numerous in sheep, goat, pig etc.

3. Circumvallate papillae: These bear numerous taste buds in pigs, and dogs and
less so in cats. In the horse and pig, the taste buds are present over the entire papillary wall
of the moat and are restricted to the bottom only in carnivores.

4. Foliate papillae: Are absent in ruminants, rudimentary without taste buds in cats
but are present in the horse, dog and rabbit.

5. Taste buds: Those in the horse are melon-shaped and in Ox and Sheep they are
Ovoid. In pigs, they are spindle-shaped and may extend into the tunica propria in the dog.
They are more or less spherical in the cat and they are poorly defined.

6. Lingual glands: Mixed glands are present in the margin of the tongue in the horse,
in the root of the tongue in Ox and horse.

In the horse, there is a fibrous cord in the middorsal region of the tongue located
between the muscles. It is composed of fibrous and elastic tissue interspersed with fibrous
and hyaline cartilage, fat and some striated muscle fibres, lingual muscles are inserted to it.

In the dog there is Lyssa, a collagenous sheath enveloping adipose tissue.

DIGESTIVE TUBE

General structure:

The digestive tube is lined internally by a continuous mucous membrane, which

begins at the mouth and ends at the anus. Beginning with the oesophagus, the alimentary
canal has a wall made up of four layers or tunics, which have some common general,
features and specific regional characteristics. The four coats are from within outwards.

1. Mucous membrane 2. Submucosa. 3. Muscularis. 4. Serosa. The general features

of these four coats are described.

1. Mucous Membrane: This consists of three layers (a) Epithelium, (b) Lamina
propria and (c) Muscularis mucosa.

a) Epithelium: The type of epithelium varies in relation to the function of that part of
the tube, it lines. In some parts, it is primarily protective, in other places it is secretory and in
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some other places it is absorptive in function. It is of the stratified squamous type in

oesophagus, rumen, reticulum and omasum of the ruminant stomach and simple columnar
from stomach to large intestine and the anus being again lined by stratified squamous
epithelium.

b) Lamina Propria: This is a layer of loose connective tissue over which lies the
epithelium. It is made up of fine interlacing fibres and contains fibroblasts and macrophages.
It is frequently infiltrated with lymphocytes and plasma cells. From the stomach to large
intestine, simple tubular glands are found in the lamina propria, isolated lymph nodules
referred to as Solitary glands may also be present. In the small intestine minute projections,
with a core of lamina propria covered by epithelium called villi are formed.

(c) Muscularis mucosae: It is a thin layer of plain muscle fibre and when present
separates the lamina propria from the submucosa. It may be arranged in two layers,
longitudinal and circular, but the two layers are not always clearly recognizable. It consists of
only isolated bundles of plain muscle fibres in the oesophagus except near its termination,
but is distinct from stomach onwards. It is absent in Rumen and Reticulum.

2. Submucosa: It consists of loose connective tissue and connects the mucous

membrane to muscularis. Glands may be present in this layer as in oesophagus and
duodenum and in ileum dense aggregations of lymphoid tissue called Peyer’s patches are
present.

3. Tunica Muscularis: The muscular coat consists of the layers of plain muscle fibres
i.e outer longitudinal and inner circular layers. In the oesophagus it is made up of v oluntary
muscle, except near its termination in some species.

4. Tunica Serosa: This consists of mesothelium (simple squamous epithelium with

an underlying layer of loose connective tissue) which is adherent to the deeper muscular
coat. Since these are reflections of the serous membranes lining the body cavities (pleura of
thoracic cavity and peritoneum of abdominal and pelvic cavities), no serous membrane will
be covering organs not contained in these cavity (oesophagus in the cervical region) and a
fibrous coat of loose connective tissue will replace the serosa.

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OESOPHAGUS

The wall of oesophagus consists of a mucous membrane lined by stratified squamous

epithelium and a muscular coat, which in the cervical region is covered by a loose fibrous
adventitia. In the thoracic region, the latter is replaced by a serous membrane.

Mucosa: It is lined by stratified squamous epithelium and is thrown into

longitudinal folds. The lamina propria is made up of closely woven collagenous fibres with
some elastic fibres interspersed in it. Its well-developed papillary body is overlaid by
superficially cornified, stratified squamous epithelium. The musclaris mucosae is made up of
longitudinal smooth muscle fibres, which may consist of a part of few muscle fibres, only in
the initial part in some species.

Submucosa: It is composed of loose network of coarse collagenous fibres, which

permit formation of longitudinal folds. Mucous glands occur in the submucosa and they are
made up of typical mucous alveoli with short ducts, which pass through the submucosa and
muscularis mucosae and open between the two adjacent connective tissue papillae. The
initial portions of the duct are lined by cuboidal epithelium but this change to a stratified
epithelium in the lamina propria. The development of the submucosal glands varies in
different parts of the oesophagus and also in different species.

Muscularis: It is made up of striated muscle with some smooth muscle at the caudal
end. It consists of two layers, which course spirally at the beginning and become distinctly an
inner and outer longitudinal layer gradually.

Fibrosa: This is composed of loose connective tissue which binds the oesophagus to
the surrounding structures in the cervical region. In the thoracic region there is serosa,
covering the muscular coat.

Bloods vessels & nerves: Vascular trunks run longitudinally in submucosa. The
branches ramify and form capillary networks in layers. Nerve cells occur in plexuses located
in submucosa and between the main layers of tunica muscularis.

Species differences:

The muscularis mucosa is complete in man but in ruminants, solipeds and cat it
consists at first only of isolated bundles. In the dog and pig it is entirely lacking in the initial
portion in the caudal half, muscle bundles made their appearance but form a continuous
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layer only in the vicinity of the stomach. The submucosal glands are present only in the
pharyangeo-oesophageal junction in the horse, ruminants and cat. In the dog, the glands
form a continuous stratum up to the stomach and in pigs the glands extend to about middle
of the oesophagus. In the pig numerous lymph nodules are present adjacent to the glands.

STOMACH

The wall of the stomach consists of a mucosa, submucosa, muscularis and serosa
tunics. The true gastric mucosa is characterized by the presence of gastric glands. Only the
stomach of man and carnivores are lined by gastric mucosa exclusively. In solipeds and
swine the oesophageal portion of the stomach bears a cutaneous mucosa (stratified
squamous epithelium). Ruminants have a stomach consisting of four compartments, three
nonglandular diverticula comprising the fore stomach and a true glandular stomach.

The forestomach consists of three compartments (1) Rumen, (2) Reticulum, and (3)
Omasum.

Forestomach of Ruminant

The wall of the fore stomach consists of a non-glandular mucous membrane, a two-
layered muscular tunic, and a serosa.

(a) Rumen:

The mucosa forms large tongue shaped or conical papillae. The mucosa has neither
gland nor lymph nodules. The stratified squamous epithelium is of the cornifying type and the
stratum corneum forms relatively thick layer on the summits of the papillae. The stratum
granulosum and stratum lucidum are more or less continuous. Cells of the stratum lucidum
often swells up to become nucleated vesicles, with a cornified wall and non-stainable
cytoplasm. The lamina propria consists of a dense, fine collagenous and many elastic fibres.
Muscularis mucosa is absent. Submucosa is loose and blends with the lamina propria
without any line of demarcation.

Tunica muscularis has two layers – an outer longitudinal and inner circular (of plain
muscle). The serosa bridges the ruminal grooves, where the subserous connective tissue is
thick with fat, nerves and blood vessels.

(b) Reticulum:

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Macroscopically the mucous membrane forms permanent folds enclosing 4 or 6 sided

spaces or cells. Smaller folds subdivide the cells. These folds bear microscopic papillae on
their sides. The folds and the papillae have a central core of connective tissue and are lined
by stratified squamous cornified type of epithelium. A dense layer of stratum corneum covers
the tip of the papillae and the folds. In very large fold a band of smooth muscle fibres occurs
running in the same direction as the fold itself. Muscularis mucosae are otherwise absent.
The two layers of muscular tunic (of plain muscle) follow an oblique course and cross at right
angles. A tunica serosa is present.

(c) Omasum:

The mucosa forms numerous folds or Omasal laminae of different sizes. These folds
or laminae are studded with numerous papillae. Each lamina includes the entire mucosa and
submucosa. The mucous membrane is lined by stratified squamous epithelium and dense
capillary nets are found under the epithelium.The muscularis mucosae are distinct and
extend into the folds and may occasionally send fibres into the papillae on the laminae. The
larger folds are composed of tissue, which resembles mucous connective tissue. The
muscular coat consists of two layers of plain muscle – an outer thin longitudinal layer and
inner thick circular layer. From the inner layer, bundles extend into the folds so that the large
folds show on section, epithelium on both sides deeper to which lies the muscularis mucosae
on each side. A thin stratum of submucosa separate the muscularis mucosae on each side
form the central band of muscular layer derived form the inner circular layer of muscularis. At
the free edge of the folds the central layer of muscle fibres fuses with the marginal thickening
of muscularis mucosae. A serosa is present.

GLANDULAR STOMACH

The wall of the stomach is composed of a mucosa, submucosa, muscularis and

serosa. Tunica mucosa consists of (1) Surface epithelium, (2) the glandular lamina propria,
(3) muscularis mucosae.

1. Mucosa: (i) Surface epithelium: It begins abruptly at the jagged border of

stratified squamous epithelium of the oesophageal mucosa. It consists of a single layer of
high columnar cells. The oval nucleus lies in the basal part of the cell. No distinct striated
border can be seen with light microscope. The surface epithelium is continued into

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depression in the gastric mucosa - the gastric pits, bottom of which show shorter and
broader cells. The glands open into the bottoms of gastric pits, which correspond to ducts.

(ii) Lamina propria: Contains gastric glands supported by delicate connective tissue
framework. The connective tissue contains fibroblasts and histiocytes and there is diffuse
infiltration with lymphocytes. There is less connective tissue between the glands than
between the pits. The gastric glands are simple branched tubular glands. Several of them
open into one gastric pit. These glands are of three types: (a) fundic (b) pyloric and (c)
cardiac glands.

(a) Fundic glands: They are distributed through the greater part of gastric mucosa.
Each gland consists of a body or main part, which ends below in a blind and dilated extremity
(the fundus) and is continued upwards into a constricted portion, the neck, which opens into
the bottom of a gastric pit. The body of the gland contains two kinds of cells- Chief and
Parietal cells.

Chief cells are cuboidal or pyramidal and contain coarse secretory (zymogen)
granules. In H and E preparations the cytoplasm of chief cells stain blue. Nuclei are spheroid
and are at the basal end, Chief cells secrete pepsin.

Parietal cells are larger than chief cells, and are oval or polygonal; the finely granular
cytoplasm stains deeply with acid dyes (eosin). Nucleus is spherical and centrally located in
the cell. These cells lie outside the Chief cells or between them. They maintain connection
with the lumen by intercellular canaliculi, which extend between the chief cells. Intracellular
canaliculi are also present. The parietal cells/ oxyntic cells secrete hydrochloric acid.

The neck of the gland is made up of mucous cells, which may be interspersed among
parietal cells. They are cuboidal with oval nuclei situated at the base of the cell.
The cytoplasm stains light blue in ordinary preparations. These cells secrete mucous, it is
believed that the mucous secreted by these cells contains the intrinsic factor, which
enhances the absorption of extrinsic factor (Vitamin B12) necessary for haemopoiesis.

Argentaffin cells: (Chromaffin cells) which show fine cytoplasmic granules, staining
black by silver impregnation technique may occur as isolated cells between the basement
membrane and chief cells. They are said to contain serotonin, a vasoconstrictor substance,
which stimulates the contraction of plain muscle.

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Fig.6.3 Histology of fundic gland

1. Surface epithelium of gastric
mucosa
2. Isthmus
3. Gastric pit
4. Mucin
5. Mucoid neck cells
6. Mucoid material
7. Parietal cells
8. Chief cells
9. Neck
10. Fundus

(b) Pyloric glands: These are coiled tubular glands. In sections mostly transverse
and oblique sections of the tubule are seen. The ducts of the glands are longer but the body
is shorter and the cells of the body resemble mucous cells. The glands secrete mainly
mucous. Between the cells of the gland, narrow eosinophilic Stohr’s cells are often found.
The gastric pits or ducts are longer (or deeper) and are lined by eosinophilic columnar cells.

(c) Cardiac glands: They occur at the transition zone, between esophagus and
stomach in simple stomached animals. In horse, it is restricted to the narrow zone close to
the Margoplicatus junction. In ruminants, cardiac glands are located close to the omaso -

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abomasal junction. These are highly branched and coiled, tubular glands. The ducts are ver y
long i.e., the pits are very deep. The body has a relatively wide lumen and lined by clear
pyramidal or cubodial cells with a basally located nucleus. They secrete mucous and these
glands pass over gradual transition into typical gastric glands.

(iii) Muscularis mucosae: Plain muscle fibres interwoven or stratified forming a thin
layer fibres extend into the lamina propria.

Fig.6.4 Structure of
gastric glands
1. Cardiac region
2. Body
3. Pyloric region
4. Intermediate region

2. Submucosa: This is made up loose connective tissue.

3. Tunica muscularis: This is made up of two layers of plain muscle fibres - an outer
longitudinal and inner circular layer. A third and innermost oblique also occurs in certain parts
of the stomach.

4. Tunica Serosa: The serous coat consists of a layer of loose connective tissue
which is covered by mesothelium.

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Blood vessels and nerves: Refer-Intestine.

Species differences: The anterior oesophageal region of the stomach in horse is

lined by a cornified stratified squamous epithelium. Behind this there is fundic gland area and
then pyloric gland area. Between the oesophageal region and fundic area, there is a narrow
cardiac area.

The abomasum of ruminant consists of cardiac, fundic and posterior pyloric gland
areas.

In the carnivores there is a fundic and pyloric area. Near the cardia there is a narrow
zone of cardiac gland area. A lamina subglandularis intervenes between muscularis
mucosae and the blind ends of the glands. In old animals, it becomes stratified into an inner
stratum granulosum rich in cells and an outer stratum compactum consisting of network of
dense, hyaline collagenous substance.

INTESTINE

Intestine consists to two parts - small and large. The small intestine is divisible into
three portions, - duodenum, jejunum and ileum, which gradually pass into one another.
Their general structure is similar although each division has certain distinctive features.

The intestinal wall consists of a mucosa, submucosa, muscularis and a serosa.

1. Tunica mucosa: This may be divided into the following layers:

(a) epithelium; (b) lamina propria, which has glands and in the small intestines forms villi and
(c) muscularis mucosae.

(a) Epithelium: Surface epithelium is simple columnar with goblet cells distributed
among the columnar cells. Columnar cells are narrow and high and reach their greatest
height on the villi. Nucleus is oval and is situated in the lower half of the cell. The cytoplasm
is finely granular and usually acidophilic. These cells shown striated free border, which
appear under light microscope as fine striations in the cytoplasm. EM studies revealed that
the striated free border is actually made up of fine closely packed cytoplasmic filaments or
processes, which are now referred to as microvilli. The microvilli are considered as an
adaptation of cell surface for better absorption of digested food material from the intestine.

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Scattered among the columnar cells are mucous secreting goblet cells. They vary in
appearance according to the amount of secretion they contain. The cell at the beginning
contains a small amount of mucin near its apical border. As secretion increases the mucous
gradually displaces the cytoplasm and the nucleus is flattened pushed the nucleus towards
the base of the cell. The distended cell has the appearance of goblet and the cytoplasm
stains faintly basophilic or not at all but can be stained intensely by special mucin stains.
The secretion is discharged through the free surface (apocrine) and goblet cells go through
cycles of secretory activity.

(b) Lamina propria has a supporting framework of reticular tissue with elastic fibres
and delicate collagenous fibres. There is diffuse infiltration with numerous lymphocytes and
in certain places isolated lymph nodules known as solitary glands may be found in the
lamina propria. In the small intestine, projections of lamina propria covered by surface
epithelium called villi occur throughout. Both in small and large intestines, the lamina propria
contain simple tubular glands, called crypts of lieberkuhn or intestinal glands.

Intestinal Villi: These structures serve to increase the intestinal surface area
available for absorption. They are narrow and elongated projections, measuring on the
average of 0.5-1mm in length and 0.2 mm in width. In the center of each villus occur a
tubular lymph space known as a lacteal and is lined by endothelium. The reticular spongy
stroma of villi contains leucocytes, fat droplets, capillaries, elastic fibres and bundles of
smooth muscle fibres. These muscle fibres originate from muscularis mucosae.

Intestinal glands or Crypts of Lieberkuhn: These are simple tubular glands found
in the lamina propria. They open between the villi and extend to the lamina propria as far as
the muscularis mucosae. They are surrounded by reticular fibres and consist of glandular
epithelium resting on a thin basement membrane. The epithelium is made-up of columnar
cells as the surface epithelium with which it is continuous but the cells of gland are shorter
and their striated border becomes less distinct until they finally disappear, in the deeper
portions of the gland. Goblet cells are more abundant in the deeper portions than in the
surface epithelium.

In the small intestine the gland fundus contains the specialized granular cells of
Paneth. These are serozymogenic having the characteristic striated chromophilic material in
the basal region and large refractile acidophilic granules apically. They show activity during

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digestion. It is believed that these cells may be producing some digestive enzymes but the
exact functional significance is not known.

Enterochromaffin or argentaffin cells also occur among the crypts of the intestinal
glands. They occur usually singly and contain specific granules in the basal part of the cell,
which are stained by silver and chromium salts. They contain serotonin but their exact
functional significance is not known.

c) Muscularis mucosae consist of smooth muscle fibres, which are in two layers
perpendicular to one another. They may interweave.

2. Submucosa – Consists of loose connective tissue and elastic fibre nets. In

contains large blood vessels, lymphatics and nerve plexus in the submucosa.

3. Tunica muscularis – This is well developed, made up of plain muscle fibres and
consists of thinner outer longitudinal layer and thicker inner circular layer. The two layers are
connected by inter-muscular connective tissue.

Blood Vessels to Stomach and Intestine

The arteries reach the gastro-intestinal trunk through the mesentery or omentum,
gives off small branches to serosa and pass through the muscular coats to submucosa
where they form an extensive plexus of large vessels called Heller’s plexus.

From this plexus, one set of branches passes outward to supply the muscular coats
and another set passes inward to the mucous membrane. Short branches supply the
muscularis mucosae, while long branches enter the lamina propria and form periglandular
capillary networks. In the small intestine, each villus receives one small artery, which passes
to the summit of the villus gives off a network of capillaries, which lie immediately under the
epithelium.

Veins correspond to the arterial branches. The capillary network from the mucous
membrane and muscular coats are drained by veins, which form a submucous venous
plexus where large veins are formed. They accompany the arteries into the mesentery.

Nerves to Stomach and Intestine

The nerve supply is from the autonomic nervous system and consists of non -
medullated sympathetic fibres and medullated preganglionic parasympathetic fibres. They
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reach the intestinal wall through the mesentery and form a myenteric plexus or plexus of
Auerbach in the inter-muscular connective tissue, in association with the ganglion cells of
the parasympathetic system. From the plexus, fine branches are given off to the muscular
coat. Fibres form this plexus also pass to the submucosa, where they form a submucosal
plexus from which fine branches are distributed to the muscularis mucosae and glands of the
mucous membrane.

Lymphatics to Stomach and Intestine

Lymph capillaries begin as blind tubules in the lamina propria in the stomach and as
the lacteal in the small intestine, which occupies the center of the long axis of each villus.
These small lymph capillaries form a plexus in deeper part of the lamina propria, from which
large vessels arises. Pass through the muscularis mucosae and forms a submucous plexus.
There is a third plexus, found in the inter-muscular connective tissue. Large vessels arise
form the submucous plexus pass through the inner muscular coat and serosa, to enter the
mesentery accompanying the arteries and veins.

Regional differences in small intestine

Duodenal Mucosa: The villi are broad spatula shaped and thin. In the epithelium,
columnar cells preponderate over goblet cells. Submucosa - Branched tubuloalveolar glands
are found in the connective tissue known as Brunner’ glands or Duodenal glands. These
glands are lined by low columnar cells, which resemble pyloric gland. The nucleus is
flattened and is located at the base of the cell and the cytoplasm is faintly basophilic. These
glands secrete mucous and the ducts, which are lined by columnar cells, open between the
villi or into the crypt of Lieberkuhn.

Species differences: The duodenal glands begin at the pylorus but its extent varies
in different animals. In man 10 to 15cm, Sheep 60 to 70cm, Goat 20 to 25cm, Pig 3 to 5
meters, Ox 4 to 5 meters and in Solipeds 5 to 6 meters. In the pig and horse lobules occur
which are purely serous or have both serous and mucous cells by side. In the horse, Ox and
Sheep, goblet cells have been found in the glandular epithelium.

Paneth cells described in the epithelium are absent in carnivores and pigs.
Enterochromaffin or Argentaffin cells are found in all domestic animals along the entire
gastro-intestinal tract.

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Jejunum Mucosa: Villi are cylindrical, elongated and filliform. Goblet cells are
proportionately more numerous than in the duodenum. Submucosa – It is made up of loose
connective tissue and does not present any special features.

Ileum Mucosa: The villi are short and tongue shaped. Goblet cells are more
numerous than in jejunum. In Submucosa – Aggregations of lymph nodules called peyer’s
patches occur in the submucosa of the ileum. They always occur on the side opposite to the
submucosa of the ileum. They always occur on the side opposite to the attachment of
mesentery. Each patch may contain ten to seventy nodules arranged side to side and the
individual nodules may be discrete and well defined by connective tissue septa or they may
coalesce except at their apices. They may extend into the lamina propria and interrupt the
muscularis mucosae. The mucous membrane over these patches may be devoid of villi, a
layer of columnar epithelium alone covering the nodules. Payer’s patches are more
prominent and numerous in young and growing animals.

Fig.6.5 Peyers patches

1. Villus
2. Peyers patch
3. Remnant of muscularis
mucosa
4. Surface epithelium

LARGE INTESTINE

The large Intestine consists of caecum, colon and rectum and the general structure of
these parts are similar. The structure of large intestine resembles that of the small intestine
except for the following differences.

Mucosa: There are no villi. Intestinal glands or crypts of Lieberkuhn are present
throughout and they are longer and straighter. The surface epithelium consists of tall
columnar cells but the Goblet cells far exceed in number than the columnar cells. The crypts
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of Lieberkuhn also contain numerous goblet cells and towards the distal portions of the
intestine the crypts appear to be lined entirely by goblet cells.

The mucous membrane of the rectum is usually thrown into a number of longitudinal
folds. At the anus, the simple columnar epithelium is replaced by stratified squamous
epithelium, which becomes continuous with the epidermis beyond the anal orifice.

The submucosa and muscularis do not present any special features. The serous coat
is absent in the terminal portion of the rectum and is replaced by a fibrous coat.

Species difference:

The large intestine of man, solipeds and pigs, possess flat bands of longitudinal
muscle known as taeniae.

In the dog and pig at recto-anal junction, tubulo-alveolar anal glands are present. In
pigs, it produces a mucous secretion and in dogs it produces a fatty secretion. In the dog,
circumanal glands occur at the site where anal mucosa becomes continuous with the skin.
These consist of a sebaceous portion, which opens through a patent duct into an adjacent
hair-follicle and a non-sebaceous portion, which exhibits no evidence of any secretory
activity.

Lateral and ventral to the anus in carnivores are the anal sacs. The wall of anal sac
is covered by stratified squamous epithelium. In the loose subepithelial layer are apocrine
tubular glands in the dog and in the cat, sebaceous glands are present in addition.
The excretory ducts of anal sacs also contain tubular and sebaceous glands.

LIVER

The Liver is the largest gland in the body and is an importan t metabolic organ of
many functions. It has also an exocrine function, the secretion of bile, which is conveyed to
the intestine by a system of ducts. The organ is invested with a connective tissue sheath
called the Glisson’s capsule most of which is covered by peritoneal serous membrane.
From the connective tissue sheath fibrous bands or septa arises and enter into the
substance of the gland and divide the gland into numerous hexagonal or polygonal units, the
hepatic lobules. These connective tissue septa from the capsule are collectively termed as
the Glisson’s capsule and the interlobular columns of connective tissue convey blood vessels
and ducts and are also referred to as the portal canals. The interlobular connective tissue is
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seen in all domestic animals and men, except in the pig and camel. In the liver of pig and
camel, the lobule is very well seen, as each hexagonal or polygonal lobule is distinctly
circumscribed by connective tissue.

At the hilus of the liver, the Glisson’s capsule forms a sheath for the hepatic artery
and portal vein which enter the liver and the hepatic duct which comes out of the liver.
The branches of these structures are conveyed by the interlobular connective tissue septa or
portal canal and these three (i.e. interlobular branches of hepatic artery, portal vein and bile
duct) are fanned at the periphery of the lobules together, forming the portal triad.

Hepatic lobule

Each lobule consists of (1) a central vein, (2) cords or laminae of hepatic cells,
radiating away from the central vein, 3) hepatic sinusoids, which converge from the periphery
toward the central vein and (4) bile canaliculi or capillaries, formed between the opposing
surfaces of hepatic cells representing the initial portions of bile ducts.

Hepatic cells: In sections they appear as cords of cells radiating away from the
central vein. The cells are arranged in regular laminae or plates extending radially form the
central vein to the periphery. The plates are one cell thick and curved. They anastomose by
interlaminar bridges and between them are broad, irregular spaces-lacunae, containing
sinusoids. Liver cells (hepatocytes) are polyhedral in shape and have a large round nucleus,
which is vesicular and show a few chromatin and one or more prominent nucleo li, cytoplasm
presents a variable appearance, depending upon the functional status. Glycogen and fat are
both dissolved in the usual preparations. Some cells have two nuclei.

Bile Canaliculi and ducts: Between the opposing faces of hepatic cells are minute
channels, the bile canaliculi. In sections they appear as fine round openings between the
two opposed hepatic cells. They are like the intercellular secretory capillaries of other glands .
They are formed by indentations or grooves on the opposing faces of two or three adjacent
cells. The bile capillaries form continuous networks within the liver plates and interlaminar
bridges. Because they are always enclosed between adjacent cells, they are as far removed
as possible from sinusoids. The hepatic cells therefore have two different physiological
surfaces through which they discharge their products. On the surface of the lobules the bile
capillaries pass by the canal of Herring into the interlobular bile duct. The canals of Herring
are lined by cuboidal epithelium. The interlobular ducts are lined by columnar epithelium. The

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interlobular bile ducts unite to form the hepatic duct which leaves the liver at the hilus. The
large ducts are lined by high columnar epithelium and have a connective tissue coat.

Hepatic Sinusoids:- The plates of liver cells are separated form one another by the
sinusoids. These are irregular tortuous blood spaces, which pursue a radial course in the
lobule and connect the ends of the interlobular portal veins with the intralobular centra l veins.
They also receive blood form the branches of hepatic artery. They fill all the interlaminar
space and anastomose irregularly.

The sinusoids are lined by irregularly alternating cells of two different kinds. One of
these is endothelial cell which has small dark nucleus. Its cytoplasm extends as a thin film
along the sinusoid. The other lining cells are larger and have processes, which appear to
extend from one wall to other of the sinusoid or extend into the sinusoid. These have large
oval vesicular nuclei and prominent nucleolus. These are referred to as stellate cells of
Kupffer. They are phagocytic cells and form a part of the reticulo endothelial system. There
is potential space between the sinusoid and hepatic cells. It is called as space of Disse,
which is traversed by a reticular fibre network. It is considered by some workers that lymph
might pass through these spaces to the lymphatic vessels in the portal canal, as within the
lobule no lymphatic capillaries are present.

The hepatic artery entering at the hilus divides into branches to supply the capsule of
Liver. Some enter the portal canals and by divisions becomes interlobular arteries which
supply capillaries to interlobular connective tissue and then become continuous with the
intralobular capillaries of portal vein. Some of these may open directly into the sinusoids.
Similarly, the portal vein enters the hilus and divides into number of branches, which pass
the portal canals as the interlobular veins. Branches of interlobular veins enter the periphery
of the hepatic lobules and break into a brush of capillaries. These intralobular capillaries
anastomose to form a sinusoidal network, the hepatic sinusoids between the laminae of
hepatic cells. These sinusoids converge towards the center of the lobule where they unite to
form the central vein. The central vein is the efferent vessels from the lobule. Several central
vein join to form a sublobular vein. Sublobular veins are found in the interlobular connective
tissue between the opposed faces of lobules. Several sublobular veins join to form a
collecting vein, which inturn join to form hepatic veins which pursue independent of the portal
venous system and open in to the posterior vena cava.

GALL BLADDER
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It is a diverticulum of the hepatic duct developmentally to store bile It is absent in

horse, elephant, rat and pigeon. Duct of gall bladder is called cystic duct, which unites with
hepatic duct and forms the bile duct.

Structure: Similar to large bile ducts. The wall consists of the following la yers.
1) Mucosa consisting of a surface epithelium and lamina propria. 2) a layer of smooth muscle
3) a perimuscular connective tissue and 4) a serous layer.

Mucosa is thrown into folds. Epithelium consists of tall columnar (which has goblet
cell in the Ox.) The lamina propria contains lymph nodules and glands. The carnivores have
few glands while ruminants have many mucous and serous glands. The muscular layer
consists of smooth muscle running in different directions, circular and oblique, with the
circular predominating, in some place regular inner circular and outer longitudinal layers can
be seen. The perimuscular connective tissue layer is usually thick and outside this is the
subserous layer of connective tissue containing blood vessels and nerves. On its free
surface it is not attached to the liver, it is covered by serosa (Peritoneum).

PANCREAS

It is a dual gland, which consists of an exocrine portion that secretes certain

digestive juices and endocrine portion that secretes hormones. Both these functions of
pancreas are carried out by distinctly different groups of cells. It is compound tubuloalveolar
gland.

The pancreas is covered by a thin layer of loose connective tissue (which does not
form a distinct capsule) from which septa pass into the gland, subdividing into many lobes.
The acini are all of the serous type and are lined by pyramidal cells resting on a basement
membrane.

The pyramidal secretory cells are arranged in a single layer. They resemble serous
acini but are distinguished by the presence of two zones. The inner zone is coarsely
granular, while the outer (basal) zone appears almost homogenous or may show fine radial
striations. The spheroid nucleus lies in outer basal zone but in such a manner as to project
into the inner zone. The nucleus contains one or two prominent acidophilic nucleoli.

The acini open into the acinar or intercalated ducts, which are fairly long and are lined
by cuboidal epithelium and have all the characteristics of intercalated ducts. It should be

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noted that in pancreas all the ducts within the lobule (inter -calated and intralobular) are lined
by cuboidal epithelium only and there are no striated ducts. The interlobular ducts are lined
by simple columnar epithelium and there is a connective coat, the thickness of which is
directly proportional to the size of the duct.

The cuboidal cells of the intercalated ducts sometimes extend into the lumen of the
acinus as a projection and these extensions appear in sections as one or more small cell
appearing within the lumen and lying in contact with apical ends of acinar cells. These are
called the centro-acinar cells of Langerhans. These differ from secreting acinar cells in
having lightly stained clear cytoplasm, devoid of secretory granules or baso philic substance.

Islets of Langerhans: The endocrine part of pancreas consists of cellular

aggregations interspersed irregularly among the acini. These cell groups are called islets of
Langerhans. Tthey have no functional communications with the duct system of the gland and
their secretions are poured directly into the blood stream. In ordinary hematoxylin eosin
preparations they appear as lightly stained cell groups, between the deeply stained acini.
The size of the islets and their number vary in different portions of the gland and also in the
different animals. In routine preparation all the cells in the islets appear to be similar, but by
special staining techniques different types have been identified.

Three types are commonly recognized 1) A or alpha cell 2) B or beta cell and 3) D or
delta cell. By Mallory azan technique - Alpha cells shows bright red granules, Beta cells
shows brown to orange granules and Delta cells shows blue granules in the cytoplasm. ‘A’
cell contains fine granules and cell membranes are distinct. ‘B’ cells contain coarse granules
and cell membranes are indistinct. With chrome-haematoxylin-phloxine method of staining-
A cells show red granules and B cell show blue granules. The relative number of A and B
cells constitute 20, 75 and 5 percent. (In the dog B and D cells produce insulin and A cells
are produce glucagon which has an antagonistic action to that of insulin. The D cells produce
somatostatin.

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Fig.6.6 Histology of pancreas

1. Exocrine acini
2. Islet cells
3. Centro-acinar cells
4. Intercalated duct
5. Capillaries

Pancreatic juices contain several proteolytic enzymes (trypsinogen), Lipase (steapsin)

and are important for normal digestive processes. The endocrine secretion, Insulin plays an
important role in carbohydrate metabolism.

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7. RESPIRATORY SYSTEM

This system serves mainly for intake of oxygen and elimination of carbon dioxide. It
may be divided into conducting and respiratory portions. The former are air conducting tubes
which connect external air with that portion of lungs where exchange of gases takes place.
The tubes are hollow and comprise nose, pharynx, larynx, trachea, bronchi of various sizes.
The bronchi divides within the lung to smaller and smaller branches, the smallest division
finally ending in alveoli, where exactly the exchange of gases takes place.

TRACHEA

Trachea is thin walled, rigid tube continuing with larynx and terminates in two chief
bronchi. The following layers are recognized: (1) Mucosa (2) Submucosa (3) a layer of
fibroelastic membrane with hyaline cartilage rings and (4) adventitia

1. Mucosa: It is lined by pseudostratified ciliated columnar epithelium with numerous

Goblet cells between the ciliated columnar cells. The basement membrane is distinct.
The lamina propria contains numerous elastic fibres and a fibro-elastic membrane may be
formed in the deeper zone, separating the lamina propria form the submusosa.

2. Submucosa: It consists of loose connective tissue and contains, mixed or mucous

glands.

3. The framework of trachea consists of series of regularly arranged C-shaped rings

of hyaline cartilage. The open segment of C is dorsal and between the free cartilages,
smooth muscle bundles (trachealis) are present. Most of the muscle fibres are arranged
transversely. A fibroelastic membrane, which blends with the perichondrium, extends
between the adjacent cartilages and also between the open ends of the cartilaginous rings
enclosing the smooth muscle bundles. This fibro elastic membrane is called the tracheal
annular ligament.

The adventitia has collagenous and elastic tissue with adipose tissue and contains
blood vessels and nerves.

Species differences: In the horse, ruminant and pig, the smooth muscle fibres lie
inside the ends of rings of cartilage, but in the dog and cat they lie outside the ring. In man, it
connects the two ends.

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LUNGS

The lungs are paired organs located in the thoracic cavity. The surface of each lung is
covered by a serous membrane (pleura). Each lung is divided into a number of lobes and
pleura, dips into interlobar tissue and covers interlobar surfaces. Each lobe is sub-divided by
interlobular tissue into lobules. Each lobule is pyramidal with apex towards the hilus and
base towards serous membrane Lobulations are visible as polygonal areas on the surface of
lungs. The lobulation is not so distinct in many animals, because there is less of interlobular
connective tissue. The septa of interstitial tissue contain bronchi, vessels and nerves. Each
lobule contains the respiratory structures arising from a terminal bronchiole.

Conducting division:

Trachea divides into two chief bronchi one for each lung, entering at the hilus, the
bronchus divides into smaller bronchi which gives rise to several orders of bronchioles.
The general structure consists of a mucosa, muscularis, a fibro-elastic membrane which
encloses a cartilage and loose collagenous peribronchial layer. Gradual changes in structure
occur, as the main bronchus, divides into smaller bronchi, several order of bronchioles till the
segment called terminal bronchiole is reached. These changes consist of the following.

The mucosa is lined by pseudo-stratified columnar cilated epithelium, in the larger

bronchi. The epithelium gradually decreases in height and the goblet cells become fewer.
The epithelium becomes simple columnar ciliated with no goblet cells in the terminal
bronchiole. The glands in the submucosa progressively decrease and finally disappear.
There is a relative increase of smooth muscle till the terminal bronchiole. The hyaline
cartilage does not occur in the form of rings as in trachea but as isolated plates in bronchi,
which also disappear, in bronchioles.

Bronchi:

Mucosa is lined by Pseudo stratified columnar ciliated epithelium with numerous

goblet cells. In submucosa mucous glands are present. Plain muscle lies inner to cartilage
plates and forms a continuous cuticular layer (unlike in the trachea). Hyaline cartilage occurs
only in the form of isolated plates.

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Fig.7.1 Parts of respiratory passage

1. Trachea
2. Primary bronchus
3. Secondary bronchus
4. Bronchiole
5. Terminal bronchiole
6. Respiratory bronchiole
7. Alveolar duct
8. Alveoli

Bronchioles (small unit):

The mucosa is lined by simple columnar ciliated epithelium with Goblet cells. No
glands occur in the submucosa. Smooth muscle forms a continuous cirular layer. Cartilage
plates are absent.

Terminal Bronchioles:

The mucosa is lined by simple columnar ciliated epithelium. No goblet cells are
present and no glands occur below the mucosa. A relatively thicker smooth muscle is
present, in the wall but cartilage is absent. The terminal bronchiole divides into two or more
respiratory bronchioles.

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Respiratory division:

From the respiratory bronchioles onwards, the structures contain alveoli in their walls
and serve both as conducting and respiratory passages. The epithelium is initially simple
cuboidal but become flattened or squamous later. No Goblet cells or glands occur in any
part. The respiratory structures consist (1) Respiratory bronchiole (2) Alveolar duct
(3) Alveolar Sac (4) Alveoli.

(1) Respiratory bronchiole: These bear alveoli in their walls and between the
alveoli, the wall is lined by simple cuboidal epithelium, which may be ciliated in the initial
portion. Smooth muscle and elastic fibres are present, though they do not form a thick layer
as in terminal bronchiole. They are arranged in a spiral direction forming a loose elastic and
contractile network. The respiratory bronchioles branch into alveolar ducts.

(2) Alveolar ducts: These are long and branch repeatedly. Their walls are formed by
alveoli, without intervening patches of cuboidal epithelium. Smooth muscle bundles are
present and are concentrated around the openings of alveoli. There is no separate
epithelium other than that of the alveoli which consists of flattened or exceedingly thin
squamous cells. The alveolar ducts open into a variable number of alveolar sacs.

(3) Alveolar sacs are extremely thin walled sacs made up of a number of alveoli. The
wall of alveolar sacs does not contain smooth muscle but a framework of elastic and reticular
fibres is present. The epithelial lining is the same as that of the alveoli.

(4) Alveoli: These are cup-shaped structures, through the wall of which interchange
of gases between the blood and air takes place. An alveolus may open into the lumen of
respiratory bronchiole, or alveolar duct or alveolar sac. Between adjacent alveoli, an inter
alveolar septum is formed.

The wall of an alveolus contains a network of fine elastic and reticular fibres and in
the meshes of this network is a rich plexus of capillaries. The lumen of the alveolus is lined
by simple squamous epithelium made up of very thin attenuated cells. The cells shows such
a thin layer of cytoplasm that except at the region of the nucleus, no outlines can be made
out in routine preparation under light microscope.

EM studies have shown that this epithelium rests on a basement membrane

wherever a capillary lies close to the alveolar wall, the basement membrane of the capillary

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endothelium and that of the alveolar epithelium fuse together. This arrangement results in a
minimum of tissue intervening between air in the alveolus and blood in the capillary, thus
facilitating gaseous exchange between the two.

In the alveolar wall, in addition to the lining epithelial cells and endothelial cells of the
capillary, connective tissue cells often referred to as septal cells are present. These
correspond to macrophages and have the potency to become actively phagocytic in function.

Blood vessels: The Capillary nets described above are derived from intralobular
arteries which are branches of pulmonary artery (functional blood) and which go along with
the terminal bronchiole. The interlobular arteries (nutrient arteries) are branches of bronchial
artery and supply bronchial wall and interstitial tissue. The satellite veins are pulmonary and
bronchial veins respectively.

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8. URINARY SYSTEM

The urinary system consists of the glandular organs the kidneys and the excretory
passages, the ureter, the Urinary Bladder and the Urethra.

KIDNEYS

The kidneys are compound tubular glands concerned with excretion of urine. It
consists of excretory or uriniferous tubules, which produce urine, con ducting tubules, which
convey urine form the kidney to the renal pelvis, and blood vessels. The kidneys are
embedded in fat and each kidney is covered by a capsule of dense collagenous fibres and a
elastic fibres.

The glandular part consists of an outer cortex and an inner Medulla. The Medulla is
in the form of renal pyramids whose bases are in contact with the cortex and apices form the
renal papillae. The glandular part surrounds a cavity adjacent to the hilus called the renal
sinus in which lies the renal pelvis, the dilated origin of ureter. The renal pelvis divides into
primary divisions or Major calyces, which in turn divide into minor calyces. Each minor calyx
receives a renal papilla and the tip of the papilla shows small openings called the area
cribrosa.

The cortex forms the outer zone but cortical tissue also projects down into the
medulla between the bases of the renal pyramids as the renal columns of Bertini.
The cortical region is subdivided into a pars convoluta or cortical labyrinth containing
convoluted tubules and glomeruli and a pars radiata or medullary rays, which are columns
of straight tubules, which radiate outward from the medulla.

The parenchyma of the kidney consists of uriniferous tubules and related blood
vessels and collecting tubules between which are a scanty amount of interstitial tissue. The
structural and functional unit of the uriniferous tubule is called a nephron. Each nephron
begins as a spherical expansion known as Bowman’s capsule, which encloses a tuft of
capillaries – the glomerulus. The Bowman’s capsule and the glomerulus together form the
Renal or Malphigian corpuscle. These are distributed in the pars convoluta of the cortex.
Apart from the Bowmans’ capsule, the nephron consists of the following segments
(1) Proximal convoluted tubule - a highly tortuous tube in the cortical labyrinth, which passes
down into the medulla as (2) the Descending limb of Henle which extends for varying

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distances into the medulla and then forms (3) the loop of Henle after which it passes up into
the cortex as (4) the Ascending limb of Henle. This reaches the renal corpuscle of its
nephron, attaches itself to its vascular pole and is then continued as the (5) Distal convoluted
tubule in the cortical labyrinth. Here ends the nephron or secretory segment of the renal
tubule and opens into arched collecting tubule in the cortical labyrinth. These arched
collecting tubules open into straight collecting tubules, located in the pars radiata of the
cortex, which pass down the medulla and reach the apex of the renal pyramid and form the
papillary ducts of Bellini, which open through the area cribrosa of renal papilla into a minor
calyx.

Structure of the parts of a nephron:

Renal corpuscle: The glomerulus consists of a number of separate capillaries

connecting an afferent arteriole with an efferent arteriole. These vessels are usually closer
together where they enter and leave the glomerulus and this end is known as the vascular
pole of the renal corpuscle.

The endothelial cells of the glomerular capillaries are extremely thin and the
cytoplasm is fenestrated. The endothelium rests on a basement membrane, which fuses with
the basement membrane of the visceral layer of Bowman’s capsule, which closely invests
the glomerulus. The Bowman’s capsule consists of two layers of epithelium, a visceral layer
closely investing the glomerulus and a parietal layer made up of simple squamous
epithelium. Between the two layers of capsule is a space where provisional urine, first formed
as glomerular filtrate collects. The visceral layer of the capsule is made up of cells whose
nuclei project into the capsular space and their cytoplasm shows numerous foot-like
processes, which are in contact with the basement membrane. Hence these cells are termed
as podocytes. The arrangement of podocytes with the fenestrated endothelium of
glomerular capillaries, leave only the basement membranes as the barrier between the blood
in the capillaries and capsular space.

The afferent arterioles of the glomerulus show special features. It lacks a distinct
tunica adventitia and there is no internal elastic membrane. In the tunica media instead of
plain muscle are myoepithelioid cells having pale staining afibrillar cytoplasm. These cells
exhibit granules demonstrable by special techniques. These are referred to as the Juxta-
Glomerular cells or apparatus (JG cells). It is now established that these JG cells secrete
a substance known as Renin, which acts on hypertersinogen in the blood to form
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hypertensin (angiotensin), which produces vaso-constriction of blood vessels and increases

blood pressure.

Neck of the tubule: Here the epithelium is of the cuboidal variety. It is a very short
segment with a diameter of 34 microns.

Proximal convoluted tubules (PCT): It is the longer and broadest portion of the
renal tubule arising as a continuation of the neck. In the pars convoluta goes towards the
surface of the cortex runs a tortuous course and then enters medullary ray to pass down into
the medulla runs straight course (termed as the straight or medullary) segment of PCT to be
continued by the descending limb of Henle.

The diameter of PCT is about 45-60 microns and is lined by high pyramidal cells
with granular cytoplasm and spherical nucleus located in the basal part. The la teral margins
of the cells interdigitate so that cell boundaries are indistinct. The apices of the cells show a
brush border (stereo-cilla), which is indistinct in ordinary preparations.

Descending limb of Henle (Thin segment of loop of Henle): It is the continuation of

the medullary segment of PCT and extends down into the medulla for varying distances.
The extent of the thin segment and length of the loop varies greatly in different tubules.

The diameter varies form 10-17 microns. It is lined by flattened squamous cells with
bulging nuclei (resembling a capillary) with a faint staining cytoplasm. The nuclei of the
opposite sides alternate.

Loop of Henle: It is present in the medulla and may show the structure of
descending or ascending segments.

Ascending limb of Henle (ALH): (thick segments of Henle’s loop) is 25-40 microns
in diameter. From the medulla, it passes up to the cortex (in pars radiata), reaches the renal
corpuscle of its nephron, attaches itself to the vascular pole and is then continued as the
distal convoluted tubule.

The ALH is lined by cuboidal cells with granular cytoplasm and spherical nuclei. Cell
outlines are distinct.

Distal convoluted tubule (DCT): It is present in pars convoluta of the cortex and
begins at the vascular pole of the Renal corpuscle and after passing a winding course

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becomes continuous with an arched collecting tubule (in the pars convoluta itself). It is much
less convoluted than the PCT and the diameter varies form 35-53 microns. It is lined by
cuboidal cells with granular cytoplasm stains intensely than the cells of PCT. The cell borders
are fairly distinct and they do not show a brush border. In cross sections cells are more
numerous, nuclei are placed close and lumen is wider than that of the PCT.

Fig.8.1 Histology of renal medulla

1. Thick segment of loop of
Henle
2. Thin segment of loop of Henle
3. Collecting duct
4. Capillary

At the Vascular pole of the Renal corpuscle, the wall of the DCT is in close contact
with afferent arteriole with JG cells. This wall of the tubule shows numerous nuclei close
together and the cells are also taller and this zone of the tubule is referred as macula Densa.
Between it and the glomerulus proper in the concavity between the efferent and afferent
arterioles is a collection of cells with pale nuclei. This group of cells is referred to as
Polkissen cells. The functional significance of Macula Densa and polkissen is not clear.

This DCT terminates the nephron or secretory segment of the renal tubule.

Arched collecting tubule: 22-60 microns in diameter. It starts form the periphery of
the medullary ray and reached the apex of pyramid. The diameter progressively incr eases.
The epithelium is cuboidal gradually increasing in height with clear lightly stained cytoplasm
and deeply stained nuclei with definite cell out lines.

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Fig.8.2 Juxta glomerular

apparatus
1. Glomerulus
2. Efferent arteriole
3. Afferent arteriole
4. Distal convoluted tubule
5. Macula densa
6. Rennin granules
7. Messangial cells

Papillary ducts of Bellini: Diameter 200-300 microns. Columnar cells with clear
basophilic cytoplasm line these. Near termination at the renal papilla the epithelium become
two layered and it may become transitional as the duct opens into a minor calyx.

Boold vessels:

The branches of renal artery at the hilus extend radialy between the pyramids as the
interlobar arteries. At the cortico-medullary junction they bend sharply and form short arches,
the arcuate or arciform arteries. The arcuate arteries give a number of interlobular arteries,
which ascend vertically through the cortical labyrinth towards the surface. From these
interlobular arteries small branches enter the glomeruli as the afferent vessel. The presence
of JG cells in the afferent glomerular arteriole has been described with the structure (Renal
Corpuscle).

The efferent glomerular arteriole after leaving the corpuscle divides into a system of
capillaries - the peritubular plexus that form a dense network around the tubules of the
cortex. The arterial supply to the medulla is furnished by efferent glomerular vessels of those
renal corpuscles, which lie close to the medulla. These vessels, the arteriolae rectae spurlae
pursue a straight course into the medulla and give rise to capillary nets, which extends to the
apex of the pyramids.

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The blood from the cortex is collected into small venules, which unite beneath the
capsule to form stellate veins. From these arise interlobular veins, which receive small veins
in the cortex and open into arcuate veins. Straight veins form the medulla open directly into
arcuate veins. From the arcuate veins large interlobular veins pass between the medullary
pyramids to form the renal vein.

Excretion of urine

The kindneys perform their functions by (1) Filtration of Blood plasma in the
glomeruli (2) selective reabsorption by the tubules of substances which the body needed to
retain (3) active excretion by the tubules of certain substances to be added to the urine and
(4) exchange of Hydrogen ions and formation of ammonia as a part of the process of acid
base regulation.

Species Differences

In the Ox and pig, numerous renal pyramids are present with the formation of minor
and major calyces. In the horse, sheep and dog the apices of the pyramids do not appear
separate and the inner part of medulla forms the renal crest (results of fusion of papillae in
the embryo), which shows numerous openings the area cribrosa. The renal crest projects
into the renal pelvis the dilated origin ureter. No minor and major calyces are formed. In the
cat the cells in the proximal convoluted tubules contain many fat droplets and a simi lar
condition is also terminal portions of the tubule in the dog.

The Excretory Passages are renal calyces pelvis, ureter, urinary bladder and urethra.

From the renal papillae urine passes into the renal pelvis of the renal pelvis of the
kidney, hence it flows through the ureter into the urinary bladder and leaves the body by way
of urethra.

Renal pelvis and calyces: These are lined by transitional epithelium resting on loose
connective tissue proper. The mucularis consists of two ill-defined layers of plain muscle and
sphincter-like arrangement of the circular layer is found in each minor calyx at the base of
each papilla and at the beginning of the ureter. A connective tissue coat containing fat cells,
large blood vessels and nerves covers the muscular coat.

In the horse: Goblet cells occur in the epithelium and tubulo-alveolar mucous gland
in the lamina propria of the renal pelvis. A distinct submucosa is present in the horse and ox.
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Ureter: It has connective tissue coat externally. The muscular coat consists of an
outer circular and an inner longitudinal layer of plain muscle fibres. In the lower third there is
an additional external longitudinal layer. The mucous membrane is thrown into folds and is
lined by transitional epithelium.

Fig.8.3 Ureter - cross section

1. Lumen
2. Transitional epithelium
3. Adipose tissue
4. Submucosa
5. Lamina propria
6. Circular muscle
7. Longitudinal muscle

Urinary bladder: The anterior part of the bladder is covered by serous membrane.
The muscular coat is very thick and consists of three layers, an outer longitudinal, middle
circular and inner longitudinal layers made up of plain muscle. The mucous membrane is
lined by transitional epithelium, which is thick when the bladder contracted and thin when
distended with urine. In the distended condition, the cells are thin, flattened and stretched
parallel to the wall resemble narrow spindle. There is no distinct submucosa. The deeper
layers of propria have a looser arrangement and so help to form thick folds.

Blood Vessels: In addition to capillary nets in the muscular coat and propria a rich
capillary plexus is seen immediately under the epithelium. Capillaries also enter the
epithelium. In the bladder there is a submucosa separated form propria, by bundles of plain
muscle running longitudinally.

Note: The Urethra will be described with the genital organs.

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9. SKIN

The function of skin is to protect the body and to serve as an organ of heat regulation,
excretion and sensation. It consists of an epithelial part - epidermis and a connective tissue
part - the dermis and other structures, such as hair follicles, sebaceous glands, sweats
gland etc. The hair, the horny portion of the hoof, nails or claws, on the digits, the horns of
cattle, etc., are also derived from the skin, by special modification and development.

The skin or Cutis is divided into the epithelial epidermis and the connective tissue
corium of dermis. The underlying loose subcutis attaches the skin to the subjacent organs.
The thickness of the skin varies considerably in different parts of the body. The relative
proportions of epidermis and dermis also vary. Hair follicles are generally present but there
are some non-hairy regions and the epidermis show considerable thickening in these
regions.

Epidermis:

This is a stratified squamous epithelium covered by a stratum corneum and rests on

the papillae of dermis. The free surface may be smooth or show elevation caused by
underlying papillae. Epidermis consists of two main layers - a deep layer stratum
germinativum or stratium Malphigi and a superficial horny layer. The deep layer is subdivided
into two and superficial into three. So starting from the corium one can distinguish the
following layers:

1. Stratum Cylindricum: consists of a layer of columnar cells resting on a thin

basement membrane.

2. Stratum spinosum: composed of polygonal cells arranged in a number of layers.

The cells show spherical nuclei and basophilic cytoplasm. They are arranged more
transversely, towards the surface. In routine preparation examined under light microscope
fine processes may be seen extending between the cells. T hese were referred to as
“intercellular bridges” and since the cell showed spines or processes projecting form their
surfaces, they were called “Prickle Cells”. EM studies have shown that these intercellular
bridges are artifacts. The membranes of the cells normally in close apposition but tend to pull
apart except at the region of desmosomes due to the shrinkage caused by technical
procedures.

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Numerous mitotic figures may be seen in this layer and both stratum cylindricum and
spinosum form the stratum germinativum.

3. Stratum granulosum: consists of a few layers of flattened rhomboid cells. These

cells show first signs of cornification in the form of basophilic granules of Keratohyalin which
increase in size and number toward surface and lend a granular app earance to this layer.
With increase of granules, nucleus disintegrates and becomes pale.

4. Stratum lucidum: is a shiny acidophilic layer of homogeneous appearance. Here

the Keratohyalin granules have liquefied to form eleidin, which is uniformly distributed
throughout the many layers of cell. The cells show neither nucleus nor cell boundaries.

5. Stratum Corneum: Consists of many layers of flat elongated cornified cells. The
material forming the cells is keratin or true horny substance. Nucleus is absent. The
superficial cells are dried horny plates, which are constantly shed off. New ones from the
deeper layers replace the cells thus lost. The superficial layer, which is constantly
desquamated, is known as stratum disjunctum.

Corium or Dermis:

The dermis varies in thickness and is composed of dense irregularly arranged

connective tissue. It contains the connective tissue fibres, fibroblasts and histiocytes. It can
be divided into a superficial or subepithelial papillary payer and a deeper reticular layer,
though the two layers blend without any distinct line of demarcation.

1. Papillary layer: This is just below the epidermis and there is a dense interweaving
of fine collagenous bundles. This layer bears on its surface contain papillary bodies, over
which the epithelium is moulded. These papillae may contain capillary loops or sensory
nerve endings of tactile sense (Tactile or Meissner’s corpuscles). Between the bases of
papillae, there may be downgrowths as inter-papillary pegs of epithelium.

2. Stratum reticulare: The reticular layer is deeper and thicker, not sharply marked
off from papillary layer. These are characterized by a network of coarse collagenous and
elastic fibres. The fibres form an extensive feltwork with rhomboid meshes, the direction of
fibres being parallel to the surface of the skin. The elastic fibres form basket -like capsular
condensations around hair-bulbs, sweat and sebaceous glands.

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Subcutis or hypodermis:

Consists of loose collagenous tissue which contains many elastic fibres cross with
each other to form a meshwork. Smaller bundles subdivide the meshes. A homogeneous
adhesive ground substance converts the fibre nets into thin membranes. The degree to
which skin can be displaced or folded depends on the development of subcutis i,e., the
thickness, length, extensibility of fibre bundles. The spaces of subcutis are filled with adipose
tissue forms a flat cushion (panniculus adiposus) which in well conditioned animals
extends to the deeply placed cutaneous muscles. There is no clear demarcation between
corium and subcutis.

HAIRS

The hairs are flexible, horny threads developed form the epidermis. They are placed
in deep narrow epidermal pits, which traverse the dermis to varying depths, and usually
extend into the subcutaneous tissue. Each hair consists of a shaft which projects above the
surface and a root embedded within the skin. At the root presents a knob-like expansion, the
hair-bulb, which is in close association on its under surface with a conical elevation of
connective tissue (form dermis) known as the papilla of the hair. Enclosing the hair root is the
hair follicle which consists of an epidermal and dermal parts.

Structure of hair:

The hair is composed entirely of epithelial cells, which are arranged in three definite
layers. (1) Medulla (2) Cortex (3) Cuticle.

Medulla: It forms the central axis of hair and consists of two or three layers of cells
which vary in appearance in different parts of the hair. In the root, cells are cuboidal with
rounded nuclei. In the shaft the cells are cornified and shrunken and the nuclei are
rudimentary or absent. The intercellular spaces are usually filled with air. The medulla is
absent from the finer, shorter hairs (lanugo) and is also absent in the hairs of scalp. If often
does not extend the whole length of the hair.

Cortex: Makes up the main bulk of the hair and consists of the several layers of cells.
In the root, it is composed of cuboidal cells with nuclei of normal appearance, in the lower
part. At higher levels the cells become progressively flattened and in the shaft the cells
become cornified, elongated and shrunken with degenerated nuclei. In coloured hair,

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pigment granules are found in and between the cells. Air accumulating in the intercellular
spaces also modifies the hair colour.

Cuticle: It is exceedingly thin and is composed of a single layer of clear cells. In the
deeper part of the root, the cuticular cells are nucleated but in the upper part of the root and
on the shaft, the cuticular cells are clear, scale like and non-nucleated. The cells overlap
giving the surface of the hair a serrated appearance.

Hair follicle: The follicle consists of the inner and outer epithelial root sheaths
derived form the epidermis and connective tissue sheaths derived form dermis.

The inner epithelial root sheath (1) Cuticle of root sheath similar to the cuticle of
hair and lies against it. (2) Huxley’s layer consisting of several rows of elongated cells, the
cytoplasm containing eleidin-like granules. (3) Henley’ layer-a row of flattened clear cells.

The outer epithelial root sheath is a direct continuation of stratum germinativum to

which it corresponds is structure.

The connective tissue sheath is derived from dermis and consists of three layers. An
inner layer of Vitreous membrane, a homogenous, narrow hyaline band, a middle layer of
connective tissue fibres arranged circularly and an outer layer of loosely arranged
collagenous fibres, running longitudinally.

All these different layers can be clearly seen only in the deeper portions of the root, a
little above the bulb but below at the level of the bulb and above in the upper part of the root
and shaft the different layers cannot be clearly demarcated.

The Hair bulb is not organized into layers but constituted a matrix of growing
multiplying cells, which superficially become transferred into the horny cells of the hair and
inner root sheath. It is in the form of bulbous thickening which surrounds the dermal papilla.
Laterally, cells of the bulb become continuous with the outer root sheath.

The papilla of the hair is similar to other papillae and consists of delicate elastic and
collagenous fires, cellular elements and capillary loops.

Muscles of the hair follicle:

The errectores pilorum, the errectors of the hair are oblique bands of smooth
muscle fibres, which arise in the subepithelial tissue and are inserted to the connective tissue
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Fig.9.1 Histology of skin

1. Stratum corneum
2. Stratum granulosum
3. Stratum spinosum
4. Basal layer
5. Arrector pili muscle
6. Duct of sweat gland
7. Hair bulb
8. Blood vessel
9. Fat cells
10. Capillary
11. Sebaceous gland
12. Sweat gland
13. Nerve ending and sensory corpuscle
14. Hair
15. Melanocyte

sheath of the hair follicle about middle of the follicle. The muscle usually arches around the
sebaceous glands, which fill the angle between the muscle and hair. The muscle is poorly
developed in certain regions like axilla, face etc. and are absent in eyelashes etc.

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The contraction of the muscle caused the hair to become more vertical to the surface
and a small depression appears in the skin at the place where the muscle is attached. This
gives rise to the so called “goose flesh”.

Colour of hair:

It is due to the presence of pigment granules found in and between the cells of the
cortex of the hair. Presence of air also modifies the colour to a certain exten t and hair in
which pigment has faded and the medulla has become filled with air appears silver white.

Skin Glands:

Sebaceous glands: These are associated with hair follicles and open into neck of
hair follicle. In certain places of the body they occur independent of hair (e.g. glans penis,
prepuce, labia vulvae, anus, external ear canal and tarsal glands of eyelids). Sebaceous
glands are simple or branched alveolar, holocrine glands. The alveolus has a basement
membrane and is filed with epithelial cells. Of these, the one nearest periphery is smaller and
cuboidal and the central ones are bigger and polygonal. The fatty secretion (sebum) in the
cytoplasm of cells is dissolved in ordinary preparations leaving honeycombed appearance.
Ducts are lined by stratified squamous epithelium.

Sweat Glands: Two kinds of these are seen, merocrine and apocrine.
The secretory portion of merocrine glands is a tubule rolled into a ball and lined by cuboidal
epithelium (coiled tubular glands). The tubule has a basement membrane. The duct has a
double layer of cuboidal cells with condensed, retractile borders. The secretory portion is
located in dermis and duct passes between the dermal papillae to open, at the free surface
of epidermis, at the sweat pore (Visible as minute pit to the naked eye). In the epidermis, the
duct has no wall of pit to the naked eye). In the epidermis, the duct has no wall of its own but
passes as spiral tunnel through the epithelium.

The secretory portions of apocrine glands are very wide. They are also coiled tubular
glands. Cytoplasm of the cell is basophilic, finely granular and contains fat droplets and
pigment. The apical portion of cells detached into the lumen during secretion. The cells after
discharge of secretion are lined with flattened nucleus. Ducts open into hair follicles above
the sebaceous glands. There is one apocrine gland per hair follicle. The secretion is fatty.

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Regional Differnce:

The epidermis is considerably thicker in the palm, sole and volar surface s of digits
and all layers are distinct. The stratum corneum is very thick and forms as major part. In
these regions the hair follicles are absent. Tactile corpuscles are numerous in the dermal
papillae in the skin, at the tip of fingers and palm.

In other region, the epidermis is thinner. All the layers are reduced and stratum
corneum and stratum germinativum are constantly present, the stratum granulosum and
lucidum being indistinct or totally absent. In the dermis the papillae are not prominent and are
less numerous. Most of the area of the skin over the body contains hair follicles.

The colour of the skin is primarily dependent upon the presence of melanin pigment
in the epidermal cells. Certain patches of skin are especially rich in pigment (nipples, a xilla,
circumanal region, scrotum, labia majora), while practically no pigment is present in some
regions (palm, sole).

Melanin is formed in specialized cells known as melanocytes. These are found in the
basal layers of stratum germinativum. The cell bodies of melanocytes appear clear, free of
granules located among the basophilic epidermal cells in routine preparations. The
processes of melanocytes are demonstrable only by special techniques, extend between the
epidermal cells. The melanin granules as they are formed are pushed to the periphery and
hence the processes of melanocytes will contain pigment but the cell body appears relatively
clear. From the processes of melanocytes, the pigment is distributed to the epidermal cells.
In white races the pigment granules occur only in the deepest cell layers and in coloured
races, the pigment is distributed in more layers, throughout the stratum germinativum.

Melanocytes stain black on treatment with Dioxy phenylalanine ( Dopa reaction) and
this reaction is attributed to the presence of the enzyme tyrosinase in the cytoplasm of
melanocytes. The formation of melanin is stimulated by ultraviolet radiation.

In the coloured races, pigmented connective tissue cells containing melanin, called
dermal chromatophores may also occur in the dermis, close to the epithelium.

Blood Vessels and Nerves:

From the larger arteries of the subcutaneous tissue, branches pass into the reticular
layer of dermis, where they form networks. From this network, branches pass into the
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papillary layer where they form secondary network. From this the papillae, hair follicles,
sebaceous glands etc., are supplied. The primary plexus supplies the sweat gland, fat
lobules etc. The venous capillaries form similar plexuses in the papillary, reticular layer and
finally in the subcutaneous tissue, form where veins arise and accompany the arteries.

The nerves to the skin are mainly sensory and sympathetic efferent supply to sweat
glands errectores pilorum and plain muscle in the walls of blood vessels. The sensory nerves
end in Pacinian corpuscles in the subcutaneous tissue, in tactile corpuscles of Meissner in
the dermal papillae and free nerve endings among the epithelial cells in the epiedermis,
pacinian corpuscles are receptors for pressure sense. Meissner’s corpuscles for tactile or
discriminatory touch and free nerve endings for pain.

Species difference:

The thickness of the skin varies with age, sex species and body region. The Ox has
the thickest skin and sheep has the thinnest skin. It is thickest on the back and on the
extensor surfaces of limbs than on the belly and flexor surfaces. The skin is very thick on the
tail of the horse, on the dewlap of the Ox and on the ventral aspect of the neck of the pig.

The wattles of the goat are appendages consisting of skin and subcutis. They include
a striated muscle, a bar of elastic cartilage, vessels and nerves.

The tip of the nose, foot pads and all muco-cutaneous junctions in all animals are
devoid of hair. The hair root is set vertically in sheep and obliquely in all other animals. The
medulla of the hair is lacking in the wool, hairs of ungulates and the hair of human scalp.

The hairs are evenly distributed in horse and ox and occur in groups in dog, cat and
pig. There are usually three hairs in a group. Of which, one is the main hair and is larger than
the other two. In carnivores, each of these three hairs is surrounded by six to twelve wool
hairs, the follicles of which branch off form the follicle of the main hair and thus a whole
bundle of hairs may project form a common follicular opening.

In tactile or sinus hairs, the connective tissue sheath is well developed and a blood
sinus lined by endothelium occurs between the outer and inner layers of dermal sheath. They
are present in the lips, nostrils and eyes.

Coarse hairs of great length occur in the mane of horse and the tail of the horse and
ox.
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Sebaceous glands are large in the horse and dog while those of the pig are
rudimentary. In ungulates two to six sebaceous glands empty into one follicle. Sebaceous
glands are largest at muco-cutaneous junctions but are absent from footpad, hooves, claws,
the planum naso labiale and teats of Ox and planum nasale of sheep, goats and carnivores.

Sweat glands: In man the merocrine glands are distributed over the entire skin
surface while the apocrine glands are restricted to a few areas such as axilla, in domestic
animal. The apocrine glands make up the majority of tubular skin glands, in sheep, pig, cat
and horse the secretory tubule is wound up (glomiform) whereas in the Ox, goat and dog, it
is serpentine. In the cat, the glands are poorly developed and are present only in a few body
areas (oral region, anus, lower jaw, foot pads).

The epidermis shows a high degree of specialization in forming the hooves, claws
and horns in the different domestic animals.

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10. GENITAL SYSTEM

Male Genital Syste m:

Consists of the testis, epididymis, vas deferens, penis and accessory genital glands.

TESTIS

Testis produces Spermatozoa (when the gland becomes mature and functional), and
male sex hormone. The testis is a cytogenous gland i.e. it produces living germ cells.
The testes are compound tubular glands. The gland is enveloped by a capsule of dense
fibrous tissue, the tunica albuginea and serous layer, which is visceral layer of tunica
vaginalis outside it. The capsule is rich in blood vessels. The capsule projects as thickening
into the gland as mediastinum testis at its posterior edge and extends through its long axis
from this mediastinum radiate septa - Septula testis towards the capsule dividing the gland
parenchyma into a number of pyramidal or cone-shaped lobuli testis, whose apices
converge towards the mediastinum testis.

Each lobule is occupied by the terminal portions of seminiferous tubules. One to three
of these occupy a lobule. They have an extremely tortuous course, rarely branch and are
called convoluted seminiferous tubules. Sperms are formed in the convoluted tubule.
The seminiferous tubule is 80-90cm. long and 100-200 microns thick with lumina of varying
sizes. They start blindly at the periphery continue as tubuli contorti and run towards the
mediastinum testis. At the mediastinum testis they become straight - the tubuli recti and
then form a system of irregular, anastomosing structure with cavernous spaces, the rete
testis. At the upper part of posterior edge 6 to 20 efferent ductules arise from the rete testis
and emerge on the surface of testis. Through many convolutions they form vascular cones,
whose apices point towards the mediastinum. The vascular cones form the head of the
epididymis. They fuse to form a single ductus epididymis. This duct is highly coiled and forms
the body and tail of epididymis. The duct gradually becomes straight to continue as ductus
deferens. Tubuli recti, rete testis, epididymis and ductus deferens forms the excretory portion
of the gland.

Seminiferous tubules:

Wall of the convoluted seminiferous tubule is composed of lamellar collagenous

connective tissue containing elastic fibres and condenses into a thin basement membrane.

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The seminiferous epithelium consists of two kinds of cells - spermatogenic cells and sertoli
cells. Spermatogenic cells through proliferation and transformation produce sperms.
They have indistinct boundaries and are not seen distinctly.

Sertoli cells are tall, irregularly columnar cells and placed perpendicular to the
basement membrane to which they are attached. They have a distal ragged edge and project
into the lumen of the tubule. Basal portion contains a light staining vesicular oval nucleus.
The cytoplasm shows a loose reticular structure. The spermatogenic cell at certain stages of
development embeds themselves into the cytoplasm of sertoli cells.

Spermatogenic cells occur in 3 to 7 layers and include several generations of cells

produced by division. The mother cells Spermatogonia make up the basal layer and by
division give rise to many generations of spermatogonia. These by growth, form
Spermatocytes which occupy a few layers. Each primary spermatocyte soon divides to form
two minute spermatids and make up the remaining layers. Spermatids become attached to
the cytoplasm of sertoli cells.

Spermatogonia have dark staining spheroid nuclei and are round Primary
spermatocytes also having round, dark staining spheroid nuclei. The secondary
spermatocytes are smaller than the primary spermatocytes and almost immediately divide
into two spermatids. The nucleus of the spermatocytes will present a variable appearance,
as the cells will be found in various stage of division. The division of primary spermatocyte i s
by meiosis or reduction division. Each primary spermatocyte gives rise to two secondary
spermatocytes by reduction or maturation division. Each secondary spermatocyte receives
only half the number of chromosomes (haploid number) present in the primary sp ermatocyte.
Each secondary spermatocyte divides into two spermatids. The spermatids are small cells
about half the size of secondary spermatocyte and have a round darkly staining nucleus.
Spermatids do not divide but by transformation form the specialized cells the Spermatozoa.

The tubuli recti and rete testis are lined by cuboidal or squamous epithelium. These
receive the sperm produced by seminiferous epithelium.

The interstitial tissue i.e. the thin collagenous tissue between the tubules contains
vessels, nerves and interstitial cells of Leydig. These cells are polyhedral. Nuclei are and
spherical with a distinct nucleolus. Their cytoplasm stains light, due to dissolution of lipoid
granules and droplets in ordinary preparations. They secrete testosterone or male hormone.

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In undeveloped testis only spematagoina are present. Only with onset of sexual
maturity several layers of sepermatogenic cells are seen.

EPIDIDYMIS

Epididymis is a highly convoluted tube to form a solid body so that in section the tube
is cut several times. The duct is enveloped by collagenous tunic, which contains circular
smooth muscle. The duct has a wide lumen and the mucosa has no folds. The epithelium is
pseudostratified columnar ciliated and consists of two types of cell-tall columnar cells with
sterocilia and small angular basal cells which do not reach the surface. Columnar cells show
elongated nuclei at different levels and contain secretory granules. Basal cells contain lipids
and so in ordinary preparations stain light. The lumen shows clumps of spermatozoa.

VAS DEFERENS

Ductus deferens or Vas deferens has a mucosa, muscularis and fibrosa. The mucosa
form longitudinal folds and is lined by pseudostratified columnar cells similar to that of ductus
epididymis. But the cells are low and stereocilia shows variable distribution, absent form
some cells and present in others.

Lamina propria is rich in elastic tissue. Muscularis is the thickest coat and shows
three layers - inner longitudinal, middle circular and outer longitudinal. The inner muscular
layer is thin. The middle circular layer is the thickest and in the stallion, bull and carnivores
the fibres of the three layers are interwoven so that the layers are less distinct. Outer fibrous
coat shows blood vessels and nerves.

ACCESSORY SEX GLANDS

These open into the urethra and include seminal vesicles, prostate and bulbourethral
or Cowper’s glands.

SEMINAL VESICLE

Seminal vesicle of bull is a thick-walled sacculated tube bent on itself several times in
a tortuous manner, to form lobules separated by heavy smooth muscular septa. The capsule
and the septa contain abundant plain muscle fibres, a feature characteristic of all accessory
glands. The mucosa is thin shows primary folds which branch into secondary and tertiary
folds. These folds project into the lumen and forms by anastomosis numerous cavities of

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different sizes separated by thin partitions of lamina propria, all open into the large central
cavity. Epithelium is pseudostratified columnar being composed of columnar cells and
irregularly shaped basal cells.

Seminal Vesicles are absent in carnivores. In horse they are true vesicles showing
mucosa, muscularis and adventitia.

PROSTATE GLAND

Prostate is a compound tubulo-alveolar gland surrounded by a capsule, which

contains abundant plain muscular tissue. From this capsule broad septa penetrate and form
a network in the interior of the gland. The septa and the abundant stroma, which separates
the alveoli, contain plenty of plain muscle fibres. The alveoli are lined by columnar
epithelium. They show folds, which subdivide the lumen into compartment. The columnar
cells may be tall or short, cytoplasm of cells is finally granular. Each lobule is traversed by an
axial duct. Ducts are lined by columnar epithelium, but near termination in the urethra it
becomes transitional.

COWPER’S GLAND

Each is a lobulated compound tubuloacinar gland covered by a fibrous

capsule overlaid by striated muscle. The stroma between the alveoli consists of fibro -elastic
tissue with a few plain muscle fibres. The secretory portions may be tubular or alveolar.
Lining epithelium shows variations depending on functional stage. It may be columnar or
cuboidal. Most of the columnar cells are of the mucous type, but the nucleus is spherical,
located at the base of cells. The cytoplasm stains basophilic but some cells may show a
granular acidiphilic cytoplasm. Smaller ducts are lined by simple columnar epithelium and
main ducts stratified columnar epithelium. Cowper’s glands are absent in carnivores.

URETHRA

Male: The urethra is a long mucous tube, which extends from the urinary bladder to
the glans penis. It consists of pelvic and extrapelvic or penile part. The pelvic part is lined
by transitional epithelium. The vas deferens, seminal vesicles, prostate glands are ope ning
into the urethra in its initial part and the ducts of bulbourethral glands open further behind.

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In the bull and boar, the pars disseminata of prostate gland forms a glandular layer in
the wall of entire pelvic urethra whereas in horse and carnivore, they are in the form of
scattered small glands.

The penile part of urethra is also lined by transitional epithelium, which changes to
stratified squamous epithelium near its termination. In the stallion and boar scattered glands
may be present as in the pelvic part.

PENIS

It consists of a body and glands. Body consists of (a) the erectile corpus
cavernosum penis, whose two crura are attached to the ischial arch. (b) corpus
cavermosum urethrae or corpus spongiosum penis surrounding urethra (c) muscles.
Glans penis forms the tip of penis.

Body of penis:

Consists of the corpora cavernosa, which have a capsule, a system of trabeculae and
the true erectile tissue. The fibrous capsule the tunica albuginea is a thick membrane of
dense collagenous tissue with elastic fibres. It gives off similarly constructed trabeculae
which are inter-connected to form a coarse frame work. The trabeculae form a median
septum between two corpora, between the trabeculae is the erectile tissue proper, which is a
vast-sponge like system of irregular vascular spaces intercalated between afferent arteries
and efferent veins.

It consists of a fine framework of lamellae and cords, continuous with the trabeculae
and albuginea and encloses communicating spaces called cavernous spaces. The spaces
are longitudinal and are best developed in the crura. The spaces are lined by endothelium,
which is continued into that of arteries and veins. They are bounded by the intercavernous
lamellae and trabeculae, which carry vessels and nerves. The spaces are largest in the
central zone of corpus cavernosum penis.

Erection of the body of penis is brought about by filling in of the cavernous spaces
with arterial blood and prevention of regress of blood through veins. Constriction of arteries
and slow emptying of cavernous spaces by contraction of smooth muscle and elastic fibres
terminate erection.

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The corpus spongiousum or corpus cavernosum urethrae have a similar structure but
the albuginea is thin and contains more elastic fibres. The trabeculae are thin l acunae are
more or less of uniform size. The urethra (penile part) passes through the corpus
spongiosum. The mucous membrane is thrown into folds and is lined by transitional
epithelium.

Species differences:

In man, three distinct bodies form penis and a median septum clearly demarcates the
two corpora cavernosa. The median septum is present only near the root of the penis in
ruminants and boar but is continuous throughout the body in dog. In stallion and cat, the
septum is not continuous.

The intercavernous framework is composed of fibroelastic tissue in ruminants and

boar and smooth muscle is also present in the horse and dog. Scattered adipose tissue
occurs in the penis of domestic animals. In the bull, the corpus cavernosum largely consists
of fibrous tissue but the erectile tissue is also present to a limited extent, which serves to
stiffen the penis rather to enlarge it.

FEMALE GENITAL SYSTEM

The female genital apparatus consists of the ovaries, oviducts, uterus, vagina and
vulva. The entire apparatus is in the scene of regularly recurring process of growth and
involution which constitutes the sexual cycle , which is interrupted only by pregnancy and old
age. Ovary like the testis is a cytogenous gland. It produces the female germ cells - ova
and also female sex hormones.

OVARY

This histological picture depends on the plane of section and phase of sexual cycle.

Ovary can be divided into two zones - cortex (zona parenchymatosa) and medulla
(zona vasculosa). It is surrounded by a tunica albuginea which itself is covered by a
germinal epithelium. In the young this germinal epithelium is a single layer of cuboidal or
columnar cells, which become flattened or squamous in adult. It is continuous with peritoneal
mesothelium. Tunica albuginea is rich in collagenous fibres. The cortical stroma is made up
of a special type of connective tissue, which is predominantly cellular. The connective tissue
cells are fusiform or spindle shaped with elongated nuclei and is placed in a network of
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delicate collagenous fibres. Elastic fibres are practically absent. Scattered in this stroma are
the glandular structure of ovary, the ovarian follicles. Each follicle consists of an ovum
surrounded by follicular epithelial cells.

In the embryo, germinal epithelium contains primordial germ cells. It forms cord-like
proliferation, which grows down into the cortex as egg tubes of Pfluger. It contains primitive
ova surrounded by a layer of small cuboidal cells. The cortical stroma grows in between
these primitive ova or oocytes, thus forming primary follicles. At birth the tunica albuginea
separates the germinal epithelium from the cortex. Thus at birth, the ovary contains all the
primary follicles and no new follicles are supposed to form after formation of tunica
albuginea.

In a mature or adult animal, the cortex will contain ovarian follicles, in various stages
of development and regressions. Smaller follicles with the ovum surrounded by a layer of
follicular cells are located near the periphery. They pass deeper into the ovary as they
become larger and develop further but the matured follicle again approaches the surface.
The follicles are described as primary, secondary, Graafian and mature Graafian follicles.

Primary Follicle: 30-35 microns in diameter. Each contains an Oocyte surrounded by

a layer of follicular cells and a basement membrane. The follicular cells are first flat later
becoming cuboidal or columnar. The follicular cells proliferate and become two layered and
then become multilayered surrounding the Ovum. These are the secondary follicles. The
Ovum increases size and may occupy an eccentric position and develop a homogenous
envelope- the zona pellucida between it and follicular cells and the stroma becomes more
distinct.

Further, growth of the follicle is characterized by proliferation of follicular cells which

form a stratified layer around the ovum. Irregular spaces appear in the follicular masses
which fuse to form a crescent shaped cavity the anturm folliculi. The cavity contains a fluid
called the Liquor folliculi. These follicles with the formation of cavity are called Graafian
follicles. The stromal cells surrounding the follicles form a sheath called Theca folliculi.

As the follicle increases in size and matures, the follicular cavity with its contained
fluid increases in size. In a mature follicle, the ovum is pressed to one side where it is
surrounded by an accumulation of follicular cells, it forms a definite projection into the cavity

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known as the germ hill or discus proligerus or cumulus oophorus. The Ovum with the
nucleus (germinal vesicle) and nucleolus (germinal spot) is surrounded by a thick

Fig.10.1 Sectional view of ovary

1. Primordial follicles
2. Primary follicles
3. Growing follicle
4. Maturing follicle
5. Cortex
6. Antrum
7. Thecae
8. Membrane granulose
9. Cumulus oophorus
10. Ovum
11. Corpus albicans
12. Tunica albuginea
13. Atretic follicle
14. Ruptured follicle
15. Medulla
16. Corpus luteum
17. Germinal epithelium
18. Regressing corpus luteum
19. Hilus
20. Blood vessel
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homogenous membrane the zona pellucida. Surrounding this are tall columnar follicular cells
arranged radially forming the corona radiata. The follicular cavity is lined by several layers
of cuboidal or polyhedral follicular cells forming the membrana granulosa. There is a
homogenous basement membrane surrounding the membrana granulosa. The theca folliculi
or the follicular sheath differentiates into a theca inter na containing numerous epithelioid,
connective tissue cells and extensive capillary network and a theca externa consisting
predominantly of fibres circularly arranged and closely packed and spindle shaped cells.

The mature Graafian follicle extends through the whole thickness of cortex and
bulges on the surface of the Ovary. At this place, the stigma and the oocyte become thin and
the follicle distended with fluid.

Ovulation is the release of the fully grown oocyte from the follicle at the surface of
the ovary. The projecting follicle ruptures at the stigma through the albuginea ova escapes
with the corona radiata cells.

Oogenesis: The ovum also undergoes maturation or reduction division, before it is

fertilized. The egg cells of primary follicles are often described as oogonia but are actually
primary oocytes, which have entered a period of rest.

It is believed that proliferation of oogonia and differentiation into primary oocytes are
completed before birth a period of rest.

The primary oocytes give rise by reduction division to a secondary oocyte (containing
the haploid number of chromosomes) and the first polar body. The polar body receives very
little cytoplasm and soon degenerates. The secondary oocyte again divides giving rise to a
ootid or mature ovum and a second polar body, which degenerates. Generally the first
maturation division occurs in the developing follicle and the second division is completed
after ovulation and at the time of or after fertilization.

Corpus luteum: After rupture, the follicular cavity closes over by healing and
becomes filled with a sero-fibrinous fluid, usually containing blood. This develops into
temporary glandular structure known as corpus luteum. The granulosa cells enlarge, the
nuclei become vesicular and stain lightly. The cytoplasm shows progressively greater
accumulation of lipid droplets and yellowish pigment granules. These are known as
granulosa lutein cells. The epitehlioid cells of theca interna also proliferate and show fatty
droplets in the cytoplasm and form theca lutein cells. These are located peripherally and
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are smaller than granulosa lutein cells and have more compact, darker staining nuclei.
A connective tissue fibre from the theca externa penetrates into the lutein mass and forms
interlacing septa, in which are numerous capillaries present. The follicular cavity is greatly
reduced and disappears as the corpus luteum enlarges further.

If the discharged ovum is not fertilized, the corpus luteum develops after an initial
period of development. This is known as cyclical corpus luteum or corpus luteum
spurium (menstruations). The cells gradually decrease in size become vacuolated and are
resorbed. The connective tissue increases in amount and finally there is transformation into a
small body of connective tissue – the corpus albicans.

It the ovum is fertilized and pregnancy occurs, the corpus luteum increases in size
and becomes corpus luteum of pregnancy or corpus luteum verum. It persists until later
months of pregnancy. After termination of pregnancy, it undergoes slow involution, becoming
transformed into corpus albicans.

Atretic follicles: Only a few of the oocytes in the follicle reach maturity. Most of the
occytes together with their follicles undergo artesia. This may set in at any stage of
development. The process involves degeneration of oocyte and invasion of the follicle by the
internal cells.

Medulla of the Ovary: There is network of fibro-elastic tissue with numerous blood
and nerve fibres.

Hormones of the Ovary: The ovary is under the direct influence of Gonadotrophic
hormones of anterior pituitary. The Follicular Stimulating Hormone (FSH) controls the
maturation of the follicle and the Leutinzing hormone ( LH) controls the formation of corpus
luteum. The maturing Graafian follicles of the Ovary produce oestrogen and the corpus
luteum produces progesterone. Since there is regular cyclical development of the follicle its
maturation and subsequent development of corpus luteum after ovulation, there is a
corresponding cyclical fluctuation in the levels of the two hormones secreted by the ovary.
These are reflected in cyclical variations in the structure of oviduct, uterus and vagina in
addition to changes in the behaviour of the animal. The cycle itself is referred to as Oestrus
cycle and considerable variations exist in different animals with regard to the duration of the
oestrus cycle, different stages of the cycle, its periodicity and the nature and extent of
changes in the various parts of the reproductive tract.

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Species difference:

In the mare, there is no division of the ovary into the cortex and medulla. The
peripheral part contains blood vessels (corresponding to the medulla of other animals)
except at the ovulatory fossa, located at the free border, which is covered by germinal
epithelium.

The number of follicle that ripens in one cycle is more or less fixed for each species.
Uniparous animals usually produce only one ovum at a time. The diameter of mature follicles
is as follows: Women- 9-12 mm., mare– upto10 mm., Cow– 20 mm., Ewe, Goat,
and Sow- 5 to 8 mm., Bitch and Cat- 2 mm.In carnivores and the sow, some follicles may
contain 2 to 6 oocytes (Giant follicles).

The lutein cells of the corpus luteum in the bitch and cat contain a little or no pigment
and the corpus luteum is pale in colour. In the ewe, goat and sow, it is grayish white or flesh
coloured because of the absence of pigment in the lutein cells. In the cow, the colour varies
from bright yellow to orange and brown as the corpus luteum develops and perists durin g
pregnancy, finally turning brick-red in a regressing corpus luteum.

OVIDUCT (Uterine tube or Fallopian tube):

Wall of the oviduct is composed of a mucosa, muscularis and serosa. Mucosa bears
a columnar epithelium, parts of which are psuedostratified in the rurminants.

The epithelium contains columnar cells some of which are ciliated, whereas others
are not ciliated. The mucosa forms large primary and small secondary folds. The lamina
propria is composed of a richly cellular connective tissue. The muscularis chiefly consists of
circularly arranged plain muscle fibres. The serosa has the usual structure of mesothelium.

The structure of oviduct shows gradual variations from its ovarian to the uteine end.
Mucosal folds are pronounced at the ovarian end gradually decrease and become negligible
at the uterine end. The muscular coat becomes thicker towards the uterine end.

The epithelium also shows changes during different stages of oestrus cycle.
The epithelium becomes taller with greater secretory activity just before and during the
oestrus period.

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UTERUS

Wall of the uterus is madeup of a mucosa, muscularis and serosa.

Muscosa or Endometrium: bears a columnar epithelium. The cells are secretory in

type. Lamina propria contains uterine glands. These glands are simple branched tubular
glands which are coiled towards their ends. The connective tissue stroma resembling
mesenchyme or embryonal connective tissue surrounds the glands. Epithelium of the glands
is simple columnar which are sometimes ciliated.

Submucosa: It is absent.

Muscularis or myometrium: consists of three layers of plain muscle fibres - an inner

longitudinal, a middle circular and a thin outer layer composed of both longitudinal and
circular fibres. The middle circular layer is thickest and forms the bulk of the muscular coat.
Between the middle and outer layers is layers is layer of connective tissue stratum vasulare
containing numerous blood vessels.

Serosa or Perimetrium: is continuous with the broad ligament and has the structure
of mesothelium.

The uterus undergoes considerable structural changes during the various stages of
oestrus cycle and during pregnancy. The cyclical changes consists of congestion, edema
and proliferation of glandular epithelium during pro-oestrus and oestrus, followed by the
secretory phase, when glands develop to the maximum become highly coiled and with high
columnar cells (during metoestrus and early diestrus). In the absence of fertilization and
Pregnancy, regressive changes appear in the mucosa accompanied by hemorrhage
(menstruation) occurring in woman, is not characteristic in animals.

During pregnancy, the endometrium undergoes considerable alterations and

contributes to the formation of placenta. The myometrium also shows pronounced
hypertrophy and hyperplasia of the muscle fibres.

Species difference:

The endometrium is line by stratified epithelium in ruminants and sow. The uterine
glands in carnivores show least branching and coiling. In the mare they are highly coiled and

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branched and in the ruminants and sow, they are much wider superficially and in the deeper
portion they are narrow, strongly coiled and branched.

In the ruminants, the mucosa bears non-glandular projections, the caruncles which
are small prominences in non-pregnant animals but develop into large, complex structures in
pregnant animals. They consist of highly cellular connective tissue covered by epithelium.
The uterine glands open on the intercaruncular mucosa.

VAGINA

The wall of the vagina consists of a mucosa, muscularis and fibrosa.

The mucosa is non-glandular and is thrown into folds. It is lined by stratified

squamous epithelium. The lamina propria consists of loose connective tissue especially rich
in elastic fibres. It contains diffuse infiltration of lymphocytes and solitary lymph nodules may
also be present. The muscularis consists of an inner thicker circular layers and a thin outer
longitudinal layer of plain muscle. The fibrosa consists of dense connective tissue with many
coarse elastic fibres and contain large blood vessels and nerves. At the anterior part there is
serosa which has the usual structure of serous membrane.

The vagina also undergoes structural changes during the various stages of oestrus
cycle. The changes are chiefly in the mucosa which increase in thickness and number of
layers of the epithelium during proestrus and oestrus. Cornification of epithelium occurs to
varying degrees in different species during oestrus, followed by desquamation at metestrus
and the epithelium being low in height at diestrus.

Species differences:

In the cow, the anterior end of the vagina has an epithelium which is lined by high
columnar cells and secrete mucous.

The cornfication of the vaginal mucosa is pronounced in rodents, carnivores and ewe
and is not distinct in cow.

MAMM ARY GLAND

The mammary gland is a compound tubuloaleolar gland. The capsule is fibroelastic

and from the capsule, tough connective tissue septa containing elastic fibres, smooth muscle
and adipose tissue enter the gland and separate the gland into lobe s and lobules. The
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amount of inter lobular connective tissue varies considerably with the functional status of the
gland, being greatly reduced in a lactating gland.

The parenchyma consists of secretory tubules with alveoli lined by short columnar or
cuboidal epithelium resting on a delicate basement membrane. The appearance of
epithelium depends on the phase of secretion. During the onset of milk formation the
granular cells are taller and show fat globules at the luminal end. After extrusion of secretion,
cells become flattened because the apical portions are cast off as secretion (apocrine gland).
The cell borders are indistinct. In a lobule, alveoli in different phase of secretion may be
seen, with high or flattened epithelium.

Ducts: (lactiferous ducts) smallest ducts are lined by secretory epithelium. Larger
ducts are lined by columnar and still larger ones by two-layered columnar epithelium.
Towards the termination there is stratified squamous epithelium.

Non-functional gland: Shows abundant interstitial connective tissue. Parenchyma

shows only ducts and few alveoli.

☺☺☺☺☺

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11. ENDOCRINE GLANDS

These are organs of internal secretion. They are rich in blood vessels. They have no
excretory ducts. They secrete specific substances called hormones.

THYROID GLAND

The gland is surrounded by a connective tissue capsule, which gives off septa of
varying thickness into the interior. These divide the organ into interconnected lobule consists
of vesicles of varying sizes called thyroid follicles. In young animals they are smaller than in
adult. Between follicles there is fine connective tissue fibres, which contain many blood
vessels and capillaries. The follicles are completely enclosed and surrounded by fine
reticular fibres. They are usually spherical or ovoid. Cells lining these are cuboidal with large
round nuclei. The cells may be flattened or squamous in distended follicles. The epithelium
rests on a thin, delicate basement membrane. The cavity of the follicle is filled with a
semifluid or gel-like substance, the thyroid colloid. The colloid generally stains acidophilic
but may be basophilic especially in activated glands. In young animals, between the follicles,
small groups of cells may be seen. These are the primitive or embryonic cells, which may
give rise to more number of follicles.

The secretion produced by the cells is stored in the vesicles as Thyroglobulin.

The thyroglobulin is hydrolysed by enzymatic action of cells and the thyroxine is secreted into
the capillaries at the base of the cells.

PARATHYROID GLAND

They are situated very close to or embedded in the thyroid glands. They have
connective tissue capsule, but when deeply embedded in the thyroid gland both may have a
common capsule. The parenchyma consists of anastomosing cords of cells between which
are numerous capillaries. Two types of cells are described.

(1) Chief cells: These are polygonal with lightly stained or clear cytoplasm. The cells
may be small, with darkly stained nuclei.

(2) Oxyphile cells: These are larger than chief cells but have smaller and darker
staining nuclei. The cytoplasm contains fine granules and stains deeply with acid dyes.

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The oxyphile cells are present only in man and ox, and only in adults or aged ones
among them. The secretion parathyroid glands (parathormone) play an important role in
regulation of calcium metabolism.

Fig.11.1 Histological
organization of
Endocrine gland
A. Cell cord sinusoid pattern
B. Thyroid follicular pattern
C. Neuro-secretory cells
1. Cell cords
2. Sinusoid
3. Follicle
4. Blood capillary
5. Colloid
6. Dendrite
7. Axons

ADRENAL or SUPRARENAL GLAND

The adrenal gland is divisible into two distinct different parts - Cortex and medulla that
are developmentally and functionally independent. It has a connective tissue capsule
containing a few elastic and smooth muscle fibres. From this many delicate trabeculae,

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 HISTOLOGY LECTURE NOTES

enclosing capillaries, penetrate into the cortex and the corticomedullary junction. The
framework of cortex consists of a network of reticular fibres.

Cortex:

The cells which occupy greater part of the gland are divided into three zones.

1. The outer thin Zona glomerulosa, next to capsule.

2. A middle thick Zona fasciculata

3. An inner moderately thick Zona reticularis which abuts on the medulla.

Zona glomerulosa consists of short columnar cells, closely packed, in groups or in
columns forming arcs immediately below the capsule. They have deep staining nuclei and
basophilic cytoplasm. (In the horse it is called as zona arcuata).

Zona fasciculata consists of columns of large polyhedral cells arranged radially in

the form of anastomosing cords extending between both the zones above and below. Cell
have central nuclei, one or two in each cell, and cytoplasm is basophilic and appear
vacuolated because in routine staining technique, the lipoid droplets in the cytopla sm gets
dissolved. The vacuolation gives a spongy appearance and hence the cells are often referred
to as spongiocytes.

Zona reticularis consists of anastomosing cells cords. The cells are cuboidal and
have deeply staining spherical nuclei. The cytoplasm stains deeply in some, lightly in others
and may contain pigment. Lipoid droplets are less in number in these cells. There are
sinusoids in the zone.

Medulla:

The cells of this part are arranged in groups or irregular cords surrounded by
sinusoids. The cells are polygonal and show numerous fine granules in the cytoplasm. The
nuclei are large, centrally located and stain lightly. The granular content of the cytoplasm
varies in different cells. The cells are arranged in such a way that one pole of the cell is
directed towards a capillary and the other pole towards a vein. When the gland tissue is fixed
in fluid containing potassium dichromate, fine brown granules are seen throughout these
cells. This is due to chromaffin reaction. The cells are therefore called pheochrome cells.
These granules are readily stained with basic stain. In addition to chromaffin cells there are
single group of sympathetic ganglion cells.
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Functions:

The adrenal cortex produces numerous substances mostly steroids and based on
their action they have been grouped into three classes. There is also some evidences to
indicate that each of these classes is apparently secreted by one particular zone of cortex.

(1) Mineralo-corticoids (aldosterone) regulates water and electrolyte balance chiefly

Na and K secreted by zona glomerulosa cells.

(2) Gluco-corticoids (cortisone and hydrocortisone) influences carbohydrate

metabolism, favour catabolism of proteins and also have anti-inflammatory, anti-allergic, anti-
rheumatic properties. They inhibit lymphoblastic activity and also antibody formation. These
are produced by cell of zona fasciculata.

(3) Sex anabolic hormones (weak androgens, 17-ketosteroids oestrogen and

progesterone)- These are concerned with the development of secondary sexual
characteristics and have an anabolic action on protein metabolism. These are produced by
cell of zona reticularis.

The Medulla secretes Epinephrine and Nor-epinephrine. These increases blood

pressure and cardiac output, and cause vasoconstriction and raise blood sugar level.

PITUITARY GLAND (Hypophysis Cerebri)

The gross structure of this gland includes two lobes - an anterior and a posterior.
Histologically the anterior lobe includes pars distalis or pars anterior and the posterior lobe
includes pars nervosa and pars intermedia. Pars tuberalis surrounds the upper part of pars
nervosa and is continuous with pars intermedia. Between the pars intermedia and pars
anterior there is a cleft.

Pars anterior:

Composed of a fine network of connective tissue which supports the tortuous and
anastomosing cell cords. Sinusoids between cell cords are numerous. The cells of these
cords are of two main types. (1) The chromophobes are characterized by pale staining non-
granular cytoplasm. (2) The choromophils are granular and larger. The chromophils are
classified as acidophils and basophils on the basis of the staining reactions of their
cytoplasmic granules. Of these the acidophil cells take eosin stain in H and E preparations.

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Chromophobes: These appear in small groups. The nuclei are surrounded by a small
amount of diffuse light staining non-granular cytoplasm. The cell boundaries are usually not
distinguishable in ordinary preparations. These are considered to be reserve or
undifferentiated cells, which give rise to granular varieties.

Chromophils: Two types of cells are recognized.

1. Acidophils - These are larger than chromophobes and their cytoplasm contains
granules, which take acid dyes. Two types of acidophils showing preference to azocarmine
named as epsilon acidophils and Orangeophils or alpha acidophils showing preference
to Orange G-stain.

2. Basophils: The cytoplasm of these contains granules of irregular size and

considerable variation has been found in their staining properties. The morphological
characters vary from large angular cells to small rounded ones. A number of staining
techniques have been used to differentiate the different types among the basophils viz.,
Periodic Acid Schiff reaction, Aldehyde-fuchsin, Aldehydethionin etc. Based on the staining
reaction to these procedures and on their morphological characters, the basophils have been
classified into Beta-1 and Beta-2 and delta-1 delta-2 types.

Functions:

Pars anterior is known to secrete a number of hormones and a number of attempts

have been made to connect the secretion of one hormone with one particular cell type. The
cell types have not been identified uniformly in all animals nor has the correlation between
secretion and cell type been conclusively demonstrated in many animals. The present
evidence indicates the following association between secretion and cell type.

Acidophils: (1) Orangeophil or alpha acidophil – Somato trophin or Growth

stimulating hormone. (2) Carminophil (epsilon acidophil) - Prolactin or Luteotrophic hormone.

Basophils: (1) Beta-1 cell – Adreno-corticotrophin (2) Beta-2 cell - Thyroid

stimulating hormone. (3) Delta cell - Luteininzing or Interstitial cell stimulating hormone
(4) Delta-2-cell - Follicle stimulating hormone.

Pars intermedia: Consists of a connective tissue framework containing (basophilic)

cells which with form dense masses. Colloidal vesicles are often seen. The cells in pars in
intermedia produce the melanocyte stimulating hormone.
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Par tuberalis: Contains numerous sinusoids and has cuboidal cells with faintly
basophilic cytoplasm. The cell form vesicles, which contain colloid. Functional significance of
this part is not known.

Pars nervosa: Has a vascular connective tissue framework housing neuroglia like
cells. The cells pituicytes show granules in their cytoplasm. The nuclei of these cells are
round or oval, with a fine chromatin network. The cytoplasm is drawn into a number of fine
processes which often end either on the walls of blood vessels or on the connective tissue
septa. In routine preparations the cytoplasm and the processes cannot be made out.
Between the cells, there is a fine mesh work of interweaving processes, which stain lightly
with eosin.

Contrary to earlier beliefs, it is now known that pars nervosa by itself does not secrete
any hormone. The nerve cells located in the supraoptic and paraventricular nuclei of the
hypothalamus are the actual secretory cells. Their secretions are conveyed by their axons to
the pars nervosa, where they are released into capillaries, adjoining axon terminations.
Accumulations of neuro secretion appear as homogenus stainable masses in pars nervosa
and are known as Herrings bodies. Two hormones have been isolated form pars nervosa.
(1) Oxytocin causes contraction of uterine musculature at the end of pregnancy
(2) Vasopressin or Antidiuretic hormone – increases blood pressure and water resorption
in kidney tubules.

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12. LYMPHOID ORGANS

All these structures contain a matrix of reticular tissue, the meshes of which are filled
with lymphocytes. In addition to formed lymphatic structures like lymph glands, tonsils,
thymus and spleen, there are diffuse lymphatic infiltrations in the body like the one in the
mucous membrane of digestive, respiratory and genital systems and conjunctiva.

TONSILS

The palatine tonsil is an egg-shaped mass, which bulges into the oral cavity. The oral
surface is covered by stratified squamous epithelium. The epithelium passes down into
the crypts in the substance of tonsil. Between the crypts, the masses of dense lymphoid
tissue arranged in folds containing the individuals follicles. At intervals, between folds of
lymphatic tissue; connective tissue septa pass up form the basal layer of connective tissue,
which separates the tonsil form the subjacent structures. Below this basal layer of connective
tissue, lie bundles of skeletal muscle fibres. Round the periphery on the luminal edge are
mucous glands – tonsilar glands.

LYMPH NODES (Lymph gland)

These occur in the course of lymphatics or lymph vessels. Lymph flows through them.
These vary in size from those just visible to naked eye to those several centimeters in
diameter. They are flattened and been-shaped with a definite hilum. They have a connective
tissue capsule containing a few elastic and smooth muscle fibres. Septa or trabeculae
detached form the capsule pass towards the interior to form shallow compartments. In the
inner zone the trabeculae anastomose and pass into the hilus. This framework divides the
node into compartments, which communicate and are filled with reticular tissue.

The reticular tissue of the node consists of reticular cells and fibres, which form a
framework. The reticular fibres are continuous with the collagen fibres at the trabeculae.
The Reticular cells have an irregular shape with processes and pale staining nuclei. These
are phagocytic reticular cells, which form part of the reticuloendothelial system. There are
also non-phagocytic undifferentiated primitive reticular cells (which may give rise to the
phagocytic reticular cells or lymphocytes).

The lymphoid tissue is arranged differently in the outer part of the node or the cortex
and the inner part of the node or medulla.

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Fig.12.1 Distribution of lymphatic tissue in the body

Cortex: The trabeculae are more or less perpendicular to the surface and divide the
cortex into a number of compartments. The trabeculae pass deeper and become continuous
with the irregularly arranged trabeculae of the medulla. In the cortical compartments the
lymphocytes are closely packed together to form cortical nodules. These are more or less
spherical discrete masses of closely packed lymphocytes but in some nodes, the nodules
may be irregular and ill defined becoming continuous laterally with adjacent nodules. The
cortical nodules often contain lighter staining central areas called germinal centers because
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 HISTOLOGY LECTURE NOTES

lymphocytes are formed in them by proliferation. In these areas, many medium sized and a
few large lymphocytes have a greater amount of cytoplasm and the undifferentiated large
lymphocytes have pale vesicular nuclei. These lightly stained centr al areas are also referred
to as secondary nodules. Active proliferation of cells in the germinal center pushes outward
to the surrounding cell and thus the primary nodule contains peripheral darker zone
consisting of closely packed small lymphocytes.

The cortical nodules are separated from the capsule and the trabeculae by channel-
like spaces called lymph sinuses through which lymph circulates. The marginal or cortical
sinus receives lymph from the afferent lymphatics. The lymph then flows down the trabec ular
sinuses to enter the sinuses in the medulla.

Medulla: The trabeculae of connective tissue are irregularly arranged and

anastomose freely. The lymphocytes do not form nodules as in the cortex, but form
anastomoising lymph cords. The medullary sinuses are wider and passing between the
lymph cords and the trabeculae.

Lymphatic vessels: The afferent lymphtics are numerous and pierce the capsule on
the convex side of the node-and open into the marginal sinus. The lymph flows through the
trabecular and medullary sinuses and from here it is collected by a plexus of vessels, which
penetrate through the capsule at the hilum, from where it flows through the efferent lymphatic
vessels.

Species differences: In pigs the arrangement of nodules in the lymph node is

entirely different. The nodules occupy the central zone and the lymph cords are distributed
peripherally. The afferent lymphatics penetrate the capsule, pass to the interior and open into
the sinuses surrounding the nodule. The efferent lymphatics leave on the convex surface of
the node at several points.

Blood vessels: The artery enters at the hilus and branches pass in the trabeculae
and give branches to capsule. Other branches pass through medullary cords to form
capillary nets in the center of the cortical nodules. The veins follow the same general course
as the arteries.

Lymph capillaries: Structure similar to blood capillaries but their diameter is greater
and they show dilations.

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Lymphatcis: structure similar to venules and veins; show a much thinner poorly
defined three layered wall. Tunica intima lacks internal elastic membrane. Tunica media is
made up of smooth muscle, elastic and collagenous tissue, T unica adventita is made up of
collagenous tissue with few muscle fibres. They have numerous valves.

HAEMOLYMPH GLANDS OR HAEMAL NODES

Occur only in ruminants and are located in the sublumbar region along the vena
cava and abdominal aorta. They are small brown to dark red organs, their size and number
varies beween the individual animals. independent of lymphatic system. Haemal nodes
develop from lymph node primordia during foetal life but lose their lymph vessels during adult
life. The structure is similar to that of lymph nodes. In healthy adult, the entire node is filled
with RBCs in the sinuses instead of lymph. Their functional signifcance is not clear, however,
they may play a role against bloodborne antigens.

Milk spots are aggregations of lymphocytes and macrophages that occur along the
course of blood vessels in omentum.

SPLEEN

Spleen has a capsule of connective tissue rich in elastic tissue and smooth muscle,
over the capsule is serosa. Trabeculae from the capsule extend into the interior and form
complicated framework by anastomoses.

The spaces within the connective tissue framework are filled with a soft sponge like
tissue known as splenic pulp. On the basis of colour differences seen in the fresh
preparation different regions have been named as white pulp and red Pulp. Both types
consist of lymphoid tissue, (i.e) reticular tissue with lymphocytes but with different
arrangement and different relations to blood vessels.

The white pulp is composed of lymph nodules distributed diffusely throughout the
substance of the spleen and passing through each nodule often eccentrically is a branch of
an artery. These spherical or ovoid lymph nodules with an eccentrically placed artery
passing through them are referred to as Splenic or Malpighian Corpuscle.

The Red pulp consists of anastomosing lymph cords or cords of Billroth between
which are numerous venous sinuses filled with blood. The lymphoid tissue itself (i.e. spaces
between reticular network and lymph cords) is infiltrated with erythrocytes, the number
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varying under different conditions. The presence of venous sinuses and infiltration with
erythrocytes are responsible for the colour of red pulp.

The framework of the splenic pulp is made up of reticular tissue. The cells of the
reticulum like those in the lymph node are of two types - Phagocytic cells belonging to the
reticulo endothelial system and non-phagocytic undifferentiated cells.

Lymphocytes are of large, medium and small sized, forming the nodules in White pulp
and lymph cords in red pulp. The Red pulp also contains monocytes, granular leucocytes,
megakaryocytes or giant cells.

The structures of spleen are best understood by following the course of blood
vessels, in their passage through the organ. The Splenic artery enters the hilum and
divides into a number of branches, which enter the trabeculae as trabecular arteries. After
a short course in the trabeculae, the trabecular artery leaves the septa and enters the splenic
pulp. The tunica adventitia of the artery is replaced by reticular tissue, which is infiltrated
with lymphocytes. The artery becomes enclosed within a sheath of dense lymphatic tissue
or a nodule forming the splenic or Malpighian corpuscle. This artery is called the central
artery or artery of white pulp, but this is actually located eccentrically within nodule. From
this artery numerous capillaries are given off which supply the lymphoid tissue and then enter
the red pulp.

The central artery after passing through the splenic corpuscle, becomes reduced in
size, looses the sheath of lymphoid tissue and enters the red pulp. As it enters the red pulp
the artery divides into a number of straight branches, which lie close together at the origin
and later diverge like the bristles of a brush. These are called penicilliform artery. Each
penicillus pursues a course through the red pulp, which can be divided, into three su ccessive
parts - pulp artery, sheathed artery and terminal arterial capillary.

The pulp artery is the longest of the three segments and has the thin coat of plain
muscle surrounding the endothelium and external to the muscle in the tissue of red pulp.
The pulp artery becomes smaller and divides into sheathed arteries.

The sheathed artery has no muscular layer and the endothelium is surrounded by a
compact mass of concentrically arranged reticular cells. These become continuous
peripherally with the reticulum of red pulp. This sheath is known as the Schweiger-Seidel
sheath and the entire arrangements are referred to as an ellipsoid.
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The sheathed artery divides into two or more terminal arterial capillaries. The
terminal arterial capillary may have conical shaped enlargement or ampulla at its termination.

The exact manner in which these capillaries terminate has been a controversial
subject. Some suggest that the empty into intercellular space of the red pulp reticulum and
that blood finds its way from the pulp spaces into the venous sinuses through perforation in
the walls of the sinuses. This is called the theory of open circulation. Other workers believe
that capillaries empty directly into venous sinuses. This is called the theory of closed
circulation. An intermediate view that some capillaries connect directly with the venuous
sinuses and that others open into pulp spaces has also been put forward.

The venous sinuses form an anastomosing plexus through out red pulp. The cells
lining the sinuses are elongated and are arranged longitudinally along the course of the
sinus. The cell bodies are enlarged at the level of the nuclei and bulge at these sites into the
lumen. Surrounding these elongated cells are circularly disposed reticular fibres, whic h
resemble the “staples” and “hoops” arrangement of a barrel. Some believe that clefts are
present between the lining cells through which blood may escape and other hold the view
that a thin homogenous membrane exists between the cells. These lining cells are believed
to be phagocytic and considered to be not a true endothelium but belonging to the
reticuloendothelial system.

The venous sinuses unite to form large pulp veins or collecting venules line by
endothelium. These venules reach the trabeculae to become trabecular veins which reaches
the hilum to form the splenic vein.

Recent studies on circulation in spleen have shown that there is a cyclical activity in
the venous sinuses, regulated by sphincters at various levels including the terminal
capillaries. The activity is in different phases and consists of

(1) Conducting phase with all sphincters open all the blood enters the sinus at the
afferent and leaves it at efferent.

(2) Filtration phase –The sphincter at the efferent end closes, blood enters the sinus
through the afferent end, the fluid passes rapidly through the sinus wall and the sinus gets
gradually distended and packed with erythrocytes and

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(3) The afferent sphincter also closes and the sinus remains distended with both
sphincters closed for some time. Then the storage phase terminates with the opening of the
efferent sphincter and the movement of erythrocytes into the collecting venules. The afferent
sphincter also opens and the conducting phase begins.

Species differences:

Mammalian spleens are of different types. The storage spleen of horse, dog and
cat is relatively large, rich in tarbeculae and muscle and poor in white pulp. The abundant
red pulp serves as storage space of blood elements and the muscular framework serves to
empty the spleen. The defense spleen is small and has few trabeculae and smooth muscle
fibres but abundant lymphatic tissue. The red pulp, the storage space of the spleen is poorly
developed in the rabbit, only moderately so in man. The ruminant and swine belong to an
intermediate type. The Ellipsoids are large in the pig and are will developed in the dog and
cat.

THYMUS

The thymus varies in size and development with the age of the individuals. It begins
to decrease in size and undergone involution with advancing age and gets gradually
replaced by fat and connective tissue.

The thymus is covered by a connective tissue capsule, which gives off numerous
septa, which divide the gland into a number of polygonal lobules. Each lobule consists of a
cortex and medulla. Often the medullary substance is continuous through several lobules.

Cortex:

It consists of a compact, dense lymphoid tissue (reticular tissue and lymphocytes).

The cells often called as thymocytes are closely packed but do not form nodules.
The reticular cells are often obscured by the loosely packed thymocytes of lymphocytes.

Medulla:

The thymocytes are less numerous and are not so closely packed together and
hence the medulla presents a lightly stained appearance in sections in contrast to the darkly
stained cortex. The Reticular tissue is more distinct.

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The medulla also contains spherical or oval bodies known as Thymic or Hassall’s
corpuscles. These are characteristic of thymus. Each corpuscle consists of a central
eosinophilic hyaline mass surrounded by concentrically arranged flattened reticular cells
which become continuous with the cells of the surrounding reticulum. The reticular tissue in
thymus is different from the reticular tissue in other organs, in that it is derive d from the
endoderm, where as the reticular tissue in other organs is derived from mesoderm.

A number of views have been held regarding the functions of thymus and it is now
considered that thymus serves as a lymphocytes producing center during foetal and early
postnatal life and the lymphocytes produced are distributed to other lymphatic organs, where
they establish colonies and multiply. The colonization of the various lymphatic tissue of the
body with lymphocytes of thymic origin begins at different times in different species. It has
also been suggested that thymus produces a Lymphocyte stimulating factor, which is
responsible for proliferation of lymphocytes in other parts of the body. The thymus has some
interrelationship with the gonads, adrenal and thyroids.

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13. NERVOUS SYSTEM

Nervous system is divided into (1) Central nervous system (CNS) and (2) Peripheral
nervous system (PNS). CNS includes the brain and spinal cord.

SPINAL CORD

Spinal cord is surrounded by spinal piamater. It is made up of outer white and inner
gray matter. Surrounding the central canal of the spinal cord is the central mass of gray
matter, the cross section of which is like ‘H’. The transverse bridge connecting the lateral
halves of this is called the gray commissure. The central canal is lined by ependyma,
enclosed by a layer of glia. The gray matter is covered all round by white matter.

The gray matter on each side consists of a dorsal horn (column), which extends
almost to the surface of the cord and a ventral horn (column), which is shorter and broader
and does not reach the surface of the cord. The portion of gray matter connecting the dorsal
and ventral horn is termed as intermediate part. The jelly-like substance called gelatinous
substance of Rolando covers the summit of the dorsal horn. In the concavity between the
ventral and the dorsal horn processes of gray matter extend into the white matter where they
interlace with the longitudinally directed fibres form the reticular formation. In the thoracic
and anterior lumbar region a small projection of gray matter on the upper part of ventral horn
is present and is known as the lateral horn.

White matter encloses the gray matter on all sides and is divided into two lateral
halves by ventral median fissure and dorsally by a dorsal median septum, which contains a
sheet of piamater. The outgoing ventral motor root and the entering dorsal sensory root into
ventral, lateral and dorsal funiculi further divide each of these halves. A bridge of white
matter - white commissure, connects the two ventral funiculi. At the entrance of the dorsal
root on either side there is dorsolateral groove or sulcus.

The gray matter of the cord consists of nerve cell and nerve fibres held together by
neuroglia. The nerve cells are in different group, (a) Motor cells -large multipolar nerve cells
upto 150 microns in size, present throughout the ventral gray (horn) column. Their dendrites
extend into the dorsal gray horn and into the ventral and lateral funiculi of white matter. T heir
axon traverses the white matter where it receives a myelin sheath and enters into formation
of ventral root of spinal nerve. (b) Column cells - make up the great majority. They are

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smaller than motor cells and distributed throughout gray matter but most abundant in the
dorsal horn. Their axons pass out into the white matter.

White matter is composed of predominantly myelinated and few non -myelinated

nerve fibres neuroglia and blood vessels, most of the myelinated nerve fibres are directed
longitudinally except in the region of the nerve roots and white commissure where they are
radial or transverse. White matter contains neither nerve cells nor dendrites.

In transverse section, spinal cord appears oval, more flattened on its ventral surface
than on the dorsal. If the section has been cut through a dorsal nerve root, a bundle of
dorsal root fibres can be seen entering the white matter dorsal and medial to the dorsal gray
horn.

Different regions of spinal cord in transection:

1. Cervical region: Section is large and oval. White matter is greater in amount than
in anyother region. Ventral gray horn is large and dorsal horn is slender.

2. Thoracic region: Section smaller and circular. Both gray horns are slender
opposite the gray commissure on both sides, is a pointed projection called lateral horn. The
lateral horn is present only in thoracic and upper lumbar segments of the cord.

3. Lumbar region: Section is larger than in thoracic region and more of gray matter
than white. Both dorsal and ventral gray horns are made of white matter.

4. Sacral region: White matter is very small in amount. Section is smallest; gray
matter is two large oval masses.

CEREBELLUM

The cerebellar hemispheres are divided by transverse fissures into lobules.

The surfaces of lobules are marked by folds (folia or lamina) running parallel to the fissure
and runs transversely to the longitudinal axis of brain. In cross section of folds, it is seen that
they give off secondary and tertiary lamina the whole producing the appearance known as
arbor vitae. The surface of secondary cerebellum is composed of gray matter – the cortex
that encloses white matter.

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Cortex:

Has an outer molecular layer (plexiform layer) with few cells and few medullated
nerve fibres, an inner granular or nuclear layer. Between these two layers a single row of
large flask shaped cells called the Purkinje cells are present. These cells give off several
antler like main dendrites which enter molecular layer and form a remarkably rich
arborization extending to the surface. The dendritic arborization is fan -shaped and extends
at right angles to the laminae. The axon is given off from the end of the cell opposite to the
dendrites and acquiring a medullary sheath passes through the granular layer to the white
matter. Purkinje cells are almost the only ones whose axons reach the white matter. Axons
of the Purkinje cells give off collaterals, which enter molecular layer and appear to terminate
there in “end buttons” upon the bodies of adjacent Purkinje cells. Cell bodies of Purkinje
cells contain concentric chromophylic bodies.

The cells in the molecular layer are either superficial stellate cells with irregular
branching dendrites and short axon or deep stellate (basket) cells. Axon of the latter extend
at right angles to the laminae for a distance of several Purkinje cells giving off to each
Purkinje cell one or more collaterals which pass towards the granular layer and envelop with
their terminal arborizations of the body and proximal non medullated portion of the axons of
Purkinje cell. Collateral’s of other basket cell axons may terminate around the same Purkinje
Cell, thus forming the basket.

The granular layer in ordinary preparations presents the appearance of closely

packed nuclei with clear spaces here and there and a few larger cells. Most of these nuclei
belong to granule cells. Their short dendrites 3 to 6 in number terminate in compact
arborizations in the granular layer. Their nonmedullated axons ascend into molecular layer
to end in varicosities. These are the parallel fibres of molecular layer and they run at right
angles to and through the dendritic expansions of Purkinje cells and their cross sections
together with terminal dendritic arborizations of Purkinje cells give the molecular layer its
punctate appearance.

In the cortex there are also the termination of afferent fibres, mossy fibres and
climbing fibres. Mossy fibres are the coarsest of the white matter while in the white matter
they bifurcate, branches going to different laminae. These main branches give off secondary
branches which enter the granular layer and arborize there. The climbing fibres pass from

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the white matter to the cells of Purknije cells. The medullated fibres of cerebellum pass
through the white matter into the granular layer, separating groups of granular cells.

The above details are best demonstrated by the Golgi method.

CEREBRUM

The Brain consists of many parts (studied in gross anatomy) having its own
histological structure only the motor area of the cerebral cortex is described below as an
example.

The cerebral cortex is the external layer of gray matter covering the convolutions and
fissures of cerebral hemispheres. It contains the bodies of nerve cells, nerve fibres,
neuroglia and blood vessels. It shows the following layers from without inwards:

1. Piamater

2. Molecular or plexiform Layer: It is poor in cells and has a framework of the

reticulum formed by the glial process and dendrites of the pyramidal cells. There are
fusiform cells directed horizontally.

3. Layer of small pyramidal cells: Contains cells with their apices directed towards
the surface. They are 10-12 microns in size. From the apex a long main dendrite passes to
the molecular layer where it forms a feltwork by its ramifications. Laterally and basally from
the cells arise dendrites. The axon arises from the base and gets myelinated. Giving off
collaterals, it passes into white matter.

4. Layer of large Pyramidal cells-giant cells of Betz: Found in the motor cortex.
Apical dendrite long and goes towards molecular layer, there are also basal dendrites. Axon
arises from the basis to the white matter giving off collaterals on its way.

5. Layer of Polymorphous cells: Cells are 5-8 microns in size, irregular and varied
in shape. Their short dendrites ramify in all directions. Their axons enter white matter.

For Histology of Sense organs refer notes on Aesthesiology.

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14. SENSE ORGANS

EYE
Cornea:

Structure of the cornea is madeup of five layers:

(1) Corneal epithelium lined stratified squamous epithelium.

(2) Lamina limitans anterior or Bowman’s membrane which is very thin and
homogeneous.
(3) Substantia propria -lamellae of connective tissue with connective tissue corpuscles-
the corneal corpuscles that are transparent.
(4) Lamina elastica or Descemet’s membrane- a thick, elastic homogeneous membrane.
This lamina divides at the periphery into three divisions, the anterior part joins the sclera;
the middle one gives attachment to the ciliary muscle and posterior division passes to the
iris forming the ligamentum pectinatum iridis.
(5) Endothelium of squamous cells; this after lining the cornea is reflected over the iris.

Chorioid:
The tissue of the chorioid is principally made up of blood vessels with numerous
pigment cells. It consists of the following layers:
(1) Lamina suprachoroidea of fibrous tissue with elastic fibres containing pigment
cells
(2)Laminavasculosaoflargebloodvessels
(3) Lamina choriocapillaris between this and the previous layer is a network of
fibroelastic tissue, the tapetum fibrosum which gives the metallic lustre and it is here only
the chorioid coat is devoid of pigment cells and blood vessels. Tapetum fibrosum is built up
of bundles of laminated areolar connective tissue cells superimposed upon the chorioid to
form a tendinous type of surface that reflects back the light passing through retina. This
causes tapetum to glisten and exhibit various types of coloration according to species and
age of the animal; (the tapetum is absent in man, monkey and pig) and
(4) Lamina basalis is thin and transparent and is made up of an inner homogeneous and an
outer elastic layer.

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Ciliary body:
It consists of three parts, ciliary ring, ciliary processes and ciliary muscle.
The ciliary ring is continuous with the anterior part of the chorioid and presents on its
inner surface, numerous fine ridges arranged in a radial manner. The ciliary processes are
formed by the plating and folding of the lamina propria and the lamina basalis of the chorioid
and are received between the corresponding folds of the suspensory ligament of the lens
The internal face of the folds is continuous with the lamina basalis there are two layers of
epithelial cells that constitute pars ciliaris retinae. The ciliary muscles constitute the outer
part of the ciliary body and lies between the sclera and ciliary processes. It forms a circular
band of unstriped muscle. They arise from the inner face of the sclera and ligamentum
pectinatum iridis close to the sclero-corneal junction and are inserted to the ciliary processes
and ring. When the muscles contract, it pulls the ciliary processes forwards, thus slackening
the suspensory ligament of the lens and allows the later to become more convex. This is the
mechanism of accommodation for near objects.
Iris:
Iris is chiefly composed of a framework of connective tissue called the stroma iridis
containing numerous pigment cells. The muscular tissue is unstriped and consists of a
sphincter pupillae around the pupil and a dilator pupillae with fibres radiating from the
sphincter to the ciliary border. The anterior surface of the iris is covered by a continuation of
the endothelium of the cornea. Beneath this is a condensation of stroma in which the cells
are close together and are filled with pigmented granules. The colour of the eye is
determined by the pigmentation of the iridic stroma. If it contains little pigment, the
pigmented epithelium on the posterior surface shows through and gives a blue coloration. In
the cat, there is diffuse yellow pigmentation of the stromal cells and so the eye has a golden
sheen. in albinos, the pigment is absent here, as elsewhere, and the iris is pink is colour.

Retina:
Histologically the retina consists of ten layers, except at the optic disc, optic papilla,
the area centralis and extreme periphery. These layers from without inward are:
1. Pigment epithelium
2. Layer of rods and cones
3. External limiting membrane
5. Outer plexiform (molecular or synaptic) layer

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6. Inner nuclear (granular) layer

7. Inner plexiform (molecular or synaptic) layer

8. Ganglion cell layer

9. Nerve fibre layer
10. Internal limiting membrane
1. The pigment epithelium is a single layer of nearly cuboidal cells, broad and
hexagonal in surface view. The outer portion of the cell contains a spherical nucleus with a
moderate amount of rounded pigment granules. The inner portion filled with dark needle
shaped pigment granules, which also extent into fine cytoplasmic processes arising from the
inner surface of the cell and reaching down between the outer membranes of the visual cells.
The melanin pigment here is a known as fuscin.
The nature and significance of the remaining layers of the retina will be understood if it
is realized that the stratification depends upon the location of three sets of neurons and their
relation to each other. These neurons, together with certain supportive elements of
ectodermal origin make up the bulk of the retina. Considering them in the order of their
conduction of nerve impulse, we find that the first neuron is represented by the
photoreceptors, the rod and cone cells. These form not only the layer of that name but
also the outer nuclear layer that consists of the nuclei and much reduced cell bodies of the
neurons. From each cell a fibre is continued inward in the outer plexiform layer, where it
makes a synaptic junction with the dendrite of a second neuron, the bipolar cell. The nuclei
of the bipolar cells lie in the inner nuclear layer; their axons pass inward to make up the inner
plexiform layer, where they effect synapse with the dendrites of the ganglion cells. The
relatively large cell bodies of the latter form the ganglion cell layer; their long axons course in
the nerve fibre layer to the optic disc, where all the axons converge to form the optic nerve.
Thus the retina consists mainly of layers of nerve cells alternating regularly with layers
formed by their processes. It is further evident that the true receptors, the rods and cones lie
further most removed and turned away from the light stimulus, which is to affect them must
pass through all the intervening layers (except at the centre of area centralis retinae). Thus
the retina is an upside-down organ. The inverted retina is characteristic of all vertebrates.

In addition to nerve cells certain supportive elements, the supporting fibres of Muller,
contribute to the formation of several layers.

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2. Layer of rods and cones lies between the external limiting membrane and the
pigment epithelium, facing the latter. The rods and cones or visual cells are the outer light
sensitive portions of this layer arranged in parallel fashion, perpendicular to the surface.
Rods: These are long and slender. Each rod shows an outer and an inner segment.
The inner segment is slightly thicker. From the inner end of the inner segment, a slender
filament passes as the rod fibre through the external limiting membrane to the outer nuclear
layer. Here it enlarges to accommodate the nucleus and is then continued as a fibre into the
outer plexiform layer. The retina from the dark-adapted eye appears purplish red. This colour
is due to rhodopsin or visual purple in the outer segments of the rod. This visual purple
bleaches in light and regenerates in the dark. Rods function under conditions of low light
intensities.
Cones: The cones are bottle or flask shaped cells, which consist of inner and outer
segments. The outer segment is small and conical. The inner segment has a bulbous form.
The inner segment is continuous with the cone fibre that passes through the external limiting
membrane. Just under this membrane the fibre enlarges to include the nucleus and is then
continued as a slightly short fibre into the outer plexiform layer. Cones function under high
intensities of illumination and are responsible for colour vision.
3. External limiting membrane : This is a thin sieve-like membrane formed by the
Muller’s fibres.
4. Outer nuclear layer: It appears to consist solely of the nuclei of the visual cells.
However, a delicate mantle of cytoplasm can be made out around each nucleus. The rod
nuclei are smaller, round in shape and stain intensely. The cone nuclei are slightly larger,
oval in shape and stain less intensely. The cone nuclei are situated close to the external
limiting membrane.
5. Outer plexiform layer: It is consists of the meshwork formed by the terminal fibres
of rods and cones and the arborizations of the dendrites of the bipolar cells. The fibres of
several rods are here related to the dendrite ending of one bipolar cell. In the case of cones,
however a single cone fibre is associated with a single bipolar cell.
6. Inner nuclear layer: It is thinner than the outer nuclear layer but is similar in
appearance. It contains the nuclei of bipolar cells, certain association neurons and the nuclei
of Muller’s fibre. The association neurons are the horizontal and amacrine cells; they
serve to interconnect the various regions of the retina. The nuclei of this layer arranged in
three zones: an outer one or horizontal cell nuclei, middle one or bipolar cell nuclei and an
inner one in which the nuclei of amacrine cells predominates.

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7. Inner plexiform layer: This consists of the processes of amacrine cells, the axons
of bipolar cells and the profusely branched dendrites of ganglion cells.
8. Ganglion cell layer: It is composed of single layer of typical multipolar ganglion
cells, among which are scattered neuroglia cells. Branches of regional blood vessels are also
seen.
9. Nerve fibre layer: It consists of the axons of ganglion cells. The non-medullated
fibres are arranged in bundles, which run parallel to the surface of the retina and converge at
the optic papilla to form the optic nerve. Between these bundles, rows of Muller’s fibres and
retinal blood vessels also pass.
10. Internal limiting membrane: it is a thin membrane of hyaline nature, formed by the
opposition of expanded bases of Muller’s fibres. It separates the retina from the vitreous
body.
Lens:
Three structural components make up the lens are capsule, anterior epithelium,
lens substance. It is enclosed in a capsule (lens capsule) that is attached by the suspensory
ligament to the ciliary processes.

i. The anterior epithelium is a single layer of cuboidal cells on the anterior surface
under the capsule. There is no posterior epithelium as these cells have given rise to the
primitive lens fibres during development. At the equator, the cells are arranged meridionally
in rows. This is the region where new lens fibres are formed constantly. The central points of
the surfaces of the lens are anterior and posterior poles and the line that connects them is
the axis of the lens.

ii. The lens substance consists of concentric lamellae of lens fibres that are colorless
and transparent.

iii. It is enclosed by a structureless highly elastic membrane, the capsule that

consists of a softer cortical substance and a dense central part, the nucleus of the lens. The
lens substance when hardened is seen to consist of concentric lamellae o f lens fibres united
by an amorphous cement substance. The lens is devoid of vessels and has no nerves.

MICROSCOPIC STRUCTURE OF THE MEMBRANOUS LABYRINTH

The walls of the membranous labyrinth are made up of outer thin tunica propria and
an inner lining epithelium. The tunica propria is composed of delicate fibrous tissue and

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stellate fibroblasts. The epithelium of the membranous labyrinth is a single layer of flat
squamous cells resting on a basement membrane.
In certain regions, the wall of the membranous labyrinth is considerably modified and
is lined by neuroepithelium. These are the macula utriculi, macula sacculi, the crista
ampullaris and the organ of Corti.
1. Maculae: These represent local thickenings of the membranous wall in the utricle
and saccule forming an elevation into the endolymphatic space. The epithelium is columnar
type in which two kinds of cells may be distinguished, the sustentacular and hair cells. The
tunica propria in the macular region is firm, thickened and is closely united to the underlying
periosteum of the bony vestibule.
The sustentacular or supporting cells are tall columnar elements resting on the
basement membrane. Their basal portions containing the oval granular nuclei and are
broader than their upper portions that pass between the hair cells. The free surface of each
sustentacular cell bears a cuticular plate. At the edge of the macula, the sustentacular cells
show a gradual transition into the simple squamous epithelium characteristic of the
remainder of membranous labyrinth.
The hair cells occupy the outer part of the epithelium. Each hair cell has the shape of
rounded flask. The large, oval and deeply staining nucleus lies towards the base of the cells.
The free surface is covered by cuticular plate, through which passes a single long tapering
process the so called hair of the hair cells. The hair is made of fine nonmotile cilia. The hairs
penetrate a membrane called the otolithic membrane , consisting of a gelatinous substance
containing a number of small bodies, the otoconia or otoliths. The otoconia are made of
calcium carbonate.
The hair cells are intimately related to the fibres of vestibular nerve. The fibres enter
the tunica propria of the macula, lose their myelin sheath and the naked axis cylinders pass
through the basement membrane and terminate around the hair cells. The maculae are
concerned with static equilibrium i.e., the position of head in space in relation to gravity.
2. Crista ampullaris: Each crista ampullaris is thickened to form a ridge, placed
transversely to the long axis of the duct. The ridge consists of connective tissue propria
containing many nerves and blood vessels and surmounted by a specialized columnar
epithelium. The columnar epithelium is similar to that of macula consisting of sustentac ular
cells and hair cells. In certain fixed preparations, the epithelium is surmounted by a tall
rounded longitudinally striated mass, the cupola. The fibres of the vestibular nerve terminate
in the epithelium of the crista in the same manner as in the macula. The cristae ampullaris

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are supposed to be concerned in dynamic equilibrium. They are stimulated by movements,

which involve any amount of rotation.
3. Cochlear duct: The wall of the bony cochlea is lined by a thin periosteal
connective tissue covered by mesenchymal epithelium in the two scalae. The vestibular
membrane is thin and homogeneous and is covered on its two surfaces by a layer of
flattened epithelium. The periosteum forming the outer wall of the cochlear duct called the
spiral ligament is greatly thickened and projects inwards as a triangular prominence termed
the crista basilaris to which the outer edge of the basilaris membrane is fixed. Immediately
above this there is concavity termed the sulcus spiralis externus, above which the
periosteum contains numerous blood vessels and is termed the stria vascularis. It is
believed that the stria vascularis secretes the endolymph of the cochlear duct.
The osseous spiral lamina consists of two plates of bone and between these are
the bipolar cells of the spiral ganglion. Minute canals pass outwards between the two plates
leading to openings in the tympanic lip of the spiral lamina, through which the peripheral
processes of the nerve cells from spiral ganglion pass to the organ of Corti on the basilar
membrane.
The basilar membrane stretches from the tympanic lip of the osseous spiral lamina
to the crista basilaris. It supports the spiral organ of Corti. The basilar membrane consists of
fine straight, unbranched fibres and the basilar fibres or auditory strings embedded in a
sparse, homogeneous ground substance. The breadth of basilar membrane varies in the
different turns of the cochlea. It is greatest at the apex, gradually diminishing towards the
base until its narrowest extent is reached in the proximal end of the basal coil.
The spiral organ of Corti: It is composed of a series of epithelial cells placed upon
the basilar membrane projecting into the endolymph of cochlear duct. It extends through the
entire length of the cochlear duct with the exception of a short distance at either end. Named
in their order from within outwards the following specialized cell types comprise the organ of
corti.
1.The border cells.

2.The inner hair cells.

3.The inner phalangeal cells.
4.The inner and outer pillar cells (inner and outer rods)
5.Outer phalangeal cells (cells of Dieters).

6.Outer hair cells

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7.Cells of Hensen
The Hensen cells are continuous with the cells of Claudius, which are cubical and
extend over the remainder of the basilar membrane to the spiral ligament. All except the hair
cells may be considered as sustentacular or supporting cells. The hair cells are intimately
related to the endings of the cochlear nerve and are neuroepithelial cells.
1. The border cells and cells of Hensen are slender columnar cells which rest on
the basilar membrane and are arranged in the inner and outer aspects of the hair cells.
2. The hair cells are columnar cells. The free surface of each has short stiff hairs,
which are in contact with the tectorial membrane. Their rounded basal ends rest on the
phalangeal cells. The inner hair cells form a single row while the outer hair cells are arranged
from 3 rows in the basal coil to 5 rows in the apical coil. The hair cell is arranged on either
side of the inner and outer pillar cells or rods of corti.
3. The phalangeal cells are arranged below the hair cells in corresponding number
of rows. Their bases rest on the basilar membrane and their upper borders are concave and
on these rest the lower ends of hair cells. The nucleus is located in the basal part of the cells
and from the upper part of the cell a slender process passes between the hair cells to the
surface and ends in a cuticular plate.
4. The pillar cells or rods of Corti: These are inner and outer rods or pillar cells and
are modified columnar cells with a broad basal part with nucleus rests on the basilar
membrane and an elongated body or pillar, which ends at the free upper extremity in an
enlarged head. The bases of the inner and outer pillar cells are farther apart but their heads
converge and articulate with each other. Thus a triangular tunnel of Corti is formed between
the two pillar cells and the basilar membrane.
Tectorial membrane: Above the cells of the organ of corti and lying in contact with
the hair cells in life is a thick, elongated gelatinous structure called the tectorial membrane.
It is usually distorted during fixation. It begins at the inner angle of the cochlear duct as a thin
layer attached to the epithelial surface on the thickened periosteum over the osseous spiral
lamina (limbus spiralis). Farther outward, the membrane becomes thicker with a
pronounced convexity on its upper surface and its lower surface is in contact with hair cells of
the organ of corti. Beyond this, it becomes thinner and ends in a rounded tip.
Nerve fibres: The peripheral processes of the bipolar cells of the spiral ganglion,
pass through the tympanic lip of the spiral lamina and reach the organ of corti. Some of the
fibres end on the inner hair cells, while others pass across the tunnel of corti and end on the

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outer hair cells. The nerve fibres form fine plexuses around the bases of the hair cells and
terminate on button like endings in contact with the surfaces of hair cells.
Functions
Cochlea: The organ of corti in the cochlea is the receptor for the sense of hearing.
Vibrations transmitted from the tympanic membrane through the chain of auditory ossicles
reach the fenestra ovalis and perilymph. It is believed that the vibrations of the basilar
membrane caused by those in perilymph stimulate the hair cells. The vibrating hair cells are
in contact with the tectorial membrane through their hairs. The stimuli are conveyed through
the spiral ganglion and the eighth cranial nerve to the brain. (For further pathways see
neurology)
Vestibule: The receptors in the macula utriculi and crista ampullaris are stimulated by
alterations of the position of head and the impulses (equilibrator or vestibular) are concerned
with the maintenance of equilibrium of the body under various conditions.

ORGAN OF SMELL
The sense of smell is carried through the olfactory nerve, which is distributed on the
nasal mucous membrane and covers the upper-fourth of the turbinate bones, septum nasi,
nasal meatuses and ethmoidal cells called olfactory area. This part of the mucous
membrane is pale-yellow in colour and is lined by epithelial cells.
The epithelium is pseudostratified columnar and is considerably thicker than that of
the respiratory region. The surface cells are of two kinds (1) sustentacular cells and (2)
olfactory cells.
The sustentacular cells are more numerous cells, each consists of three parts:
(a) A superficial portion, which is broad and cylindrical and contains pigment and
granules, arranged in longitudinal rows. The cells have well marked striated thickened
borders, which unite to form the so called membrana limitans olfactoria.
(b) A middle portion, which contains an oval nucleus. As the nucleus of these cells
all lie in the same plane, they form a distinct narrow band, which is known as the zone of
oval nuclei.
(c) A thin filamentous process, which extends from the nuclear portion down
between the cells of the deeper layers. This process is irregular and pitted by the pressure of
the surrounding cells. It usually forks and apparently anastomoses with processes of other
cells to form a sort of protoplasmic reticulum.

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The olfactory cells lie between the sustentacular cells. Their nuclei are spherical lie
at different levels and most of them are more deeply placed than the sustentacular cells.
From the nuclear portion of each cell a delicate process extends to the surface, where it ends
in several minute hair like processes. From the opposite pole of the cell a longer process
extends centrally which has a centripetal nerve fibre of one of the olfactory bulb. The
olfactory cell is thus seen to be the bipolar nerve cell with a short peripheral and a long
central process and is not a neuroepithelial cell but an analogue of spiral ganglion cell
being only example of peripherally placed ganglion cell found in certain lower animals.
The basal cells are placed between the nuclear parts of the olfactory cells and the
basement membrane. These are small nucleated elements with irregular branching
cytoplasm of which anastomoses with that of the neighboring basal cells and of the
sustentacular cells to form the peculiar protoplasmic reticulum already mentioned.
The basement membrane is not well developed.
The stroma consists of loosely arranged collagenous fibres, delicate elastic fibres
and connective tissue cells. Embedded in this stroma are numerous simple branched tubular
glands, the glands of Bowman. Each tubule consists of a duct, a body and a fundus. The
secretary cells are larger and irregular and contain a yellowish pigment, which with that of the
sustentacular cells is responsible for the colour of the olfactory mucosa.

ORGAN OF TASTE
The tongue is the organ of taste. It owes this property to the taste buds, which are
situated in the foliate, fungiform and vallate papillae in the mucouse membra ne of the
tongue, the oral face of epiglottis and anterior pillars of soft palate.
The taste buds are oval flask shaped masses, which occupy the recesses in the
epithelium The base of each taste bud is wider and rests on the corium of the mucous
membrane. It receives a branch from the combined sensory nerve formed by the union of the
chorda tympani of the VII nerve and the gustatory branch of the V nerve in the anterior two -
thirds of the organ and from the lingual branch of the IX nerve if in the posterior-third. The
apex of each bud is narrow and communicates with the mouth cavity by a small gustatory
pore.
Each bud presents a small opening towards the surface, the taste or gustatory
pore. The taste bud consists of fusiform supporting cells grouped around central taste or
gustatory cells. The taste cells are neuroepithelial cells. The external end of each taste cells
bears a filament, the gustatory hair which projects through the gustatory pore. The internal

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end of the taste cells ends in a fine process, which may be single or branched. Taste buds
are innervated by gustatory fibres of glossopharyngeal, facial and vagus nerves.

ORGAN OF TOUCH

Skin

The skin or the common integument is the protective covering of the body and is
continuous at the natural orifices with the mucous membranes of digestive, respiratory and
urinogential tracts. It acts as a sense organ concerned with the reception of pain,
temperature (warmth and cold), touch (coarse touch and tactile) and pressure sensations.
There are various types of nerve endings in the skin, some in the epidermis, others in the
dermis, around roots of hairs etc., which serve as the receptors for the above sensations.
These include:

1. Free nerve endings in the epidermis for pain.

2. Merkel’s discs, meissner’s corpuscles, peritrichial plexus etc, for touch and tactile
sensations.
3.Ruffini’s corpuscles for warmth and Krause’s corpuscles for cold.
4.Pacinian corpuscles for pressure sensation.

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15. HISTOLOGICAL TECHNIQUE

The technique includes methods of preparation and examination of tissues and cells.

Methods of Preparation:

(a) Dissociation or teasing of tissues: In this method, details of individual cells are
well seen but their anatomical relationships are destroyed. The organ or tissue is dissociate d
in some fluid medium and examined under microscope. The fluid medium used has the
property of dissolving the intercellular cement substance. At the same time fixes and
preserves the cell structure (Eg.) Muller’s fluid, 1% Normal saline, 30% Alcohol.

(b) Smear technique : Smears can be made by ‘impression-smear’ method or

spreading with platinum loop or crushing and spreading material with another side.
Preparation can be fixed, stained and examined (e.g. Examination of blood smears). They
can be cleared and mounted using a mountant.

(c) Sectional methods: Here anatomical relations are preserved and maintained.
Extremely thins sections are cut. Finished preparation is mounted in a medium with high
refractive index to enable its detailed study. Medium that is used for embedding tissues
depends upon technique employed and type of material to be sectioned. Most common
medium is paraffin wax. In certain cases celloidin, and occasionally gelatin are used.

(i) Paraffin Technique : has the widest application. The tissue is embedded in a
block of paraffin wax with melting point of 58o to 60oC after subjecting them to various
processes which render them permeable to molten paraffin wax.

(ii) Frozen Sections: Unfixed or fixed tissue is frozen on a special microtome with
carbon dioxide. This method is especially useful when rapid results are required or when
demonstrating materials soluble in alcohol or cleaning agents like xylol.

(d) Vital staining: Some living cells take up certain stains (vital stains) which colour
certain elements in cells (e.g. mitochondria). Some cells like phagocytes engulf microscopic
coloured particles, which are then visible inside the cells (e.g. elements of R.E. system).
Vital staining is subdivided into

Supravital staining: This refers to staining living cells outside the body. Janus green
and neutral red stains cytoplasmic organells like mitochondria. Living cells are treated with

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very dilute solutions of the stains. The cells are alive when they take up the stains but die
subsequently as the stains are toxic.

Intravital staining: Dyes like Trypan blue, Lithium carmine or Indian ink are injected
subcutaneously or intravenously and tissues are collected from the site of injection (when
subcutaneously injected) or pieces of organs containing phagocytic cells are prepared and
processed by the usual methods. The dye will appear as fine granules in the cytoplasm of
phagocytic cells.

Methods of examination of tissue:

1. Using optical microscope with direct or transmitted illumination.

2. Using polarizing microscope.

3. Using dark ground illumination.

4. Using phase-contrast microscope to examine living cells and tissues without

staining.

5. Using electron microscope for obtaining magnification upto 1, 00,000 times.

Of these, the most common method is using ordinary optical microscope with direct or
transmitted illumination.

Fixation:

Fixation is the method of preserving the cell and tissue constituents as close to the
living status as possible. Adequate and complete fixation is the foundation of good
histological preparation. It is essential that tissues are fixed as soon as possible after death
or removal from the body. The amount of fixative (that is an agent used for fixation) should
be 15 to 20 times the bulk of tissue to be fixed. Tissues or sectioning should be sufficiently
thin to be adequately fixed throughout in a reasonable time. The best thicknes s for routine
use is 3 to 5 µm.

Aims and effects of fixation:

Fixation is required to prevent putrefaction and autolysis to preserve and harden, to

solidify colloid materials and to aid visual differentiation of structures. Certain fixatives have
effects on subsequent staining and this should be remembered. Fixation also renders cells
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insensitive to hypotonic and hypertonic solutions. Fixation alters the refractive indices of
various components of cell and tissue into varying degrees so that details are easily seen.
Fixative must be used in solutions either with normal saline or distilled water.

Common fixatives employed:

1. Formalin is the most common fixative used in 5 to 10% strength in normal saline.
2. Mercuric chloride in combination with other fixatives.
3. Osmium tetroxide (Incorrectly named Osmic acid) to fix fat and lipids used as a 2 to
5% aqueous solutions).
4. Chromic acid: used as a 2% solution preserves carbohydrates.
5. Potassium dichromate used a 3% aqueous solution. This is another common fixative
used in combination with others.
6. Picric acid used as a saturated aqueous solution in combination with others.
7. Acetic acid-used in varying strengths in combination with other.
8. Trichlor-acetic acid used with others in varying strengths.

Type of Fixatives:

Generally the fixatives are named after their inventors

Formal Saline 5-10%

Heidenhain’s Susa

Bouin’s fluid

Zenker’s fluid and Zenker formaldehyde

Flemming’s fluid

Carnoy’s fluid

Helly’s fluid

Of this Numbers 1, 2, 3 and 4 are micro anatomical fixatives that are accurately
preserve the relations of tissue layers and large aggregates of cells to one another and are
useful in routine work. Numbers 5,6 and 7 are cytoplasmic and nuclear fixatives. In general,
fixatives are either (1) precipitant fixatives (that is they precipitate or coagulate colloidal
constituents (e.g. mercuric chlorides, alcohol, picric, chromic acid) or (2) non -precipitant
fixatives (that is they fix proteins not by chemical combination but by denaturing them which

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involves a change in the shape of protein molecule and results in making it insoluble (eg.)
Formalin, Osmium tetroxide and Potassium dichromate.

Paraffin method of preparing tissue:

The Scheme given below refers to procedures commonly employed for routine
preparations of histological sections. It should be noted that procedures may have to be
considerably modified or additional new process introduced when other fixatives are used or
special methods followed. The method given below deals with fixations using formal saline ,
embedding in paraffin and staining by Heamatoxylin and eosin.

1. Fixation in 100% Formal saline

2. Washing in water

3. Dehydration in ascending grades of (a) 50% (b) 70% (c) 90% (d) Absolute alcohol

(usually three changes)

4. Clearing commonly in xylol (sometimes benzene, toluene, or cedar wood oil).

5. Inclusion in paraffin embedding oven at 56 o to 60oC

(a) Permeation with paraffin three changes

(b) Casting of tissues (orientation and cooling the mass).

6. Sectioning.

a) Sectioning (microtomy)

b) Fixing section to slide

7. Staining

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