Class 6.

EMBRYONIC DEVELOPMENT
The life of a metazoan, produced by sexual reproduction, begins with a zygote. Formation of its
complete body from the zygote is its development. The entire process of development is called
embryogenesis (ontogenesis). The study of development is called embryology.

DEVELOPMENTAL PERIODS

The entire life history of sexually reproducing metazoans is divisible into two periods:
1) Embryogenic or Pre-natal Period. It is passed within the egg or the uterus (womb) of the mother.
2) Post-embryonic or Post-natal Period. This period extends from hatching or birth up to death.

DEVELOPMENTAL BIOLOGY

It is a branch of biology which deals with the various aspects of development of individuals during
their entire life history.

PHASES OF EMBRYONIC DEVELOPMENT

Sexually reproducing metazoans include the following five basic processes or phases in their embryonic
development.

1. Gametogenesis. It is the formation of gametes. It includes spermatogenesis (formation of sperms)

and oogenesis (formation of ova).
2. Fertilization. Union of male and female gametes to form zygote is called fertilization.
3. Cleavage and Blastulation. Cleavage includes a series of successive and rapid mitotic divisions which
transform single celled zygote into a multicellular body consisting of many cells called blastomeres. The
cleavage ends with the formation of a blastula. Formation of the blastula is called blastulation.
4. Gastrulation. Formation of gastrula is called gastrulation. Gastrula is a reorganized form of blastula.
The gastrula consists of germ layers. The latter give rise to complete individual.
5. Organogenesis. It is a process by which different organs and organ-systems are formed from the germ
layers. In fact, the cells of the germ layers give rise to the tissues, organs and organ systems.

FERTILISATION

Fertilization - is the fusion of gametes to give rise to a new individual organism or offspring and initiate
its development.
With each ovarian cycle, a number of primary follicles begin to grow, but usually only one reaches full
maturity, and only one oocyte is discharged at ovulation. At ovulation, the oocyte is in metaphase of the
second meiotic division and is surrounded by the zona pellucida and some granulosa cells.
Before spermatozoa can fertilize the oocyte, they must undergo:
1) Capacitation, during which time a glycoprotein coat and seminal plasma proteins are removed from
the spermatozoon head
2) The acrosome reaction, during which acrosin- and trypsin-like substances are released to penetrate
the zona pellucida.
During fertilization, the spermatozoon must penetrate:
1. The corona radiata
2. The zona pellucida
3. The oocyte cell membrane

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Figure 1. The three phases of oocyte penetration.
In phase 1, spermatozoa pass through the corona radiata barrier; in phase 2, one or more spermatozoa
penetrate the zona pellucida; in phase 3, one spermatozoon penetrates the oocyte membrane while losing
its own plasma membrane. Inset shows normal spermatocyte with acrosomal head cap.

Phase 1: Penetration of the Corona Radiata. Of the 200 to 300 million spermatozoa normally
deposited in the female genital tract, only 300 to 500 reach the site of fertilization. Only one of these
fertilizes the egg. It is thought that the others aid the fertilizing sperm in penetrating the barriers protecting
the female gamete. Capacitated sperm pass freely through corona cells.
Phase 2: Penetration of the Zona Pellucida. The zona is a glycoprotein shell surrounding the egg that
facilitates and maintains sperm binding and induces the acrosome reaction. Both binding and the acrosome
reaction are mediated by the ligand ZP3, a zona protein. Release of acrosomal enzymes (acrosin) allows
sperm to penetrate the zona, thereby coming in contact with the plasma membrane of the oocyte.
Permeability of the zona pellucida changes when the head of the sperm comes in contact with the oocyte
surface. This contact results in release of lysosomal enzymes from cortical granules lining the plasma
membrane of the oocyte. In turn, these enzymes alter properties of the zona pellucida (zona reaction) to
prevent sperm penetration and inactivate species-specific receptor sites for spermatozoa on the zona
surface. Other spermatozoa have been found embedded in the zona pellucida, but only one seems to be able
to penetrate the oocyte.
Phase 3: Fusion of the Oocyte and Sperm. Cell Membranes The initial adhesion of sperm to the oocyte
is mediated in part by the interaction of integrins on the oocyte and their ligands, disintegrins, on sperm.
After adhesion, the plasma membranes of the sperm and egg fuse. Because the plasma membrane covering
the acrosomal head cap disappears during the acrosome reaction, actual fusion is accomplished between
the oocyte membrane and the membrane that covers the posterior region of the sperm head.

As soon as the spermatocyte has entered the oocyte (Fig. 2):

1. The head of the sperm separates from the tail, swells, and forms the male pronucleus

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 2. The oocyte finishes its second meiotic division immediately after entry of the spermatozoon and
forms the female pronucleus (one of the daughter cell is second polar body, the other daughter
cell is the definitive oocyte. It becomes female pronucleus)
3. The zona pellucida becomes impenetrable to other spermatozoa.
As a result of the release of cortical oocyte granules, which contain lysosomal enzymes, the
oocyte membrane becomes impenetrable to other spermatozoa, and the zona pellucida alters its
structure and composition to prevent sperm binding and penetration (polyspermy).

Figure 2. A. Oocyte immediately after ovulation, showing the spindle of the second meiotic division. B.
A spermatozoon has penetrated the oocyte, which has finished its second meiotic division. Chromosomes
of the oocyte are arranged in a vesicular nucleus, the female pronucleus. Heads of several sperm are stuck
in the zona pellucida. C. Male and female pronuclei. D,E. Chromosomes become arranged on the spindle,
split longitudinally, and move to opposite poles. F. Two-cell-stage.

In overview, fertilization can be described as the following steps (Fig. 3):

• Sperm Capacitation.
• Sperm-Zona Pellucida Binding.
• The Acrosome Reaction.
• Penetration of the Zona Pellucida.
• Sperm-Oocyte Binding.
• Egg Activation and the Cortical Reaction.
• The Zona Reaction.
• Post-fertilization Events.

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 Figure 3. Fertilization.

The main results of fertilization are as follows:

1. Restoration of the diploid number of chromosomes, half from the father and half from the mother.
Hence, the zygote contains a new combination of chromosomes different from both parents.
2. Determination of the sex of the new individual. An X-carrying sperm produces a female (XX) embryo,
and a Y-carrying sperm produces a male (XY) embryo. Therefore, the chromosomal sex of the embryo
is determined at fertilization.
3. Initiation of cleavage. Without fertilization, the oocyte usually degenerates 24 hours after ovulation.

CLEAVAGE (SEGMENTATION)

Definition
Cleavage is a series of rapid mitotic divisions of the zygote which convert the single celled zygote into a
multicellular structure called blastula (blastocyst).

Process
About thirty hours after fertilization, the newly formed zygote divides into two cells the blastomeres, in
the upper portion of the Fallopian tube. This is the first cleavage. The next division occurs within forty hours
after fertilization. The third division occurs about three days after fertilization. During these early cleavages,
the young embryo is slowly moving down the Fallopian tube towards the uterus. Embryo divide again to form
a 16-cell morula (mulberry). Inner cells of the morula constitute the inner cell mass, and surrounding cells
compose the outer cell mass. The inner cell mass gives rise to tissues of the embryo proper, and the outer cell
mass forms the trophoblast, which later contributes to the placenta.
About the time the morula enters the uterine cavity, fluid begins to penetrate through the zona pellucida into
the intercellular spaces of the inner cell mass. Gradually, the intercellular spaces become confluent, and
finally, a single cavity, the blastocele, forms. At this time, the embryo is a blastocyst. Cells of the inner cell
mass, now called the embryoblast, are at one pole, and those of the outer cell mass, or trophoblast, flatten

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and form the epithelial wall of the blastocyst. The zona pellucida has disappeared, allowing implantation to
begin.
When the blastomeres divide completely the cleavage is called holoblastic (Figure 4).

Figure 4. Some stage in segmentation of the fertilized ovum.

Significance of Cleavage
Cleavage brings about:
1) the distribution of the cytoplasm of the zygote, amongst the blastomeres,
2) increased mobility of the protoplasm, which facilitates morphogenetic movements necessary for cell
differentiation, germ layer formation and the formation of tissue and organs,
3) the restoration of the cell size and the nucleocytoplasmic ratio characteristic of the species.
4) Unicellular zygote is converted into multicellular embryo.

Differences between cleavage and ordinary mitosis

1) Cleavage results in decrease of the cell size whereas mitosis maintains the cell size.
2) In ordinary mitosis the cell division is followed by a period of growth while cleavage is not followed
by growth of blastomeres.
3) There is a greater synthesis of DNA during cleavage.
4) Divisions are quick in cleavage.
5) 5) Consumption of oxygen is greatly increased in cleavage.

Function of the Zone Pellucida

The trophoblast has the property of being able to stick to the uterine (or other) epithelium and its cells
have the capacity to eat up other cells. They can, therefore, invade and burrow into tissues with which they
come in contact. As the embryo travels down the uterine tube, and the uppermost part of the uterine cavity, it
is prevented from 'sticking' to the epithelium by the zona pellucida. During this time it receives nutrition,
partly from the substances stored within the ovum (e.g. yolk), and partly by diffusion from the uterine
secretions. By the time a blastocyst is formed, it is necessary for the embryo to acquire additional sources of
nutrition. This is achieved when the blastocyst 'sticks' to the uterine endometrium, and gets implanted
in it. However, before this can happen, it is necessary for the zona pellucida to disappear. The zona
pellucida disappears soon after the morula reaches the uterine lumen. Thus, the function of the zona
pellucida is to prevent implantation of the blastocyst at an abnormal site.

Blastocyst
At the next stage of development, which produces an embryo with about sixty-four cells, a cavity is
formed within the cell mass (Figure 5.). This cavity is called blastocoel and the embryo is termed the
blastocyst which is composed of an outer envelope of cells, the trophoblast and inner cell mass.

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Figure 5. Formation of blastocyst

The trophoblast includes the blastocoel and the inner cell mass. The latter is the precursor of the
embryo. It means the inner cell mass gives rise to the embryo. The cells of the trophoblast, or trophoectoderm
(trophos - to feed) form the placenta and fetal (embryonic) membrane. The zona pellucida disappears in the
blastocyst.

Implantation of the blastocyst

The process of the attachment of blastocyst with the uterine wall of the mother is called implantation.
It occurs after 7 days of fertilization. The trophoblast is destined to form the extra embryonic mesoderm and
the chorionic villi embedding into the uterine mucosa, the endometrium.
The trophoblast initially consists of two layers of cells, an inner cytotrophoblast composed of individual
cells and an outer syncytiotrophoblast composed of a multinucleate syncytium. The trophoblastic cells
digest and phagocytize materials in the endometrium that were stored prior to implantation. One of the major
contributions of the trophoblast is its secretion of chorionic gonadotropin probably by the cells of the
syncytiotrophoblast. Chronic gonadotropin has properties similar to those of luteinizing hormone and
luteotropic hormone of the pituitary gland. It prevents the corpus luteum from involuting. The secretion of
the chorionic gonadotropin begins the day the trophoblast is embedded in the endometrial lining.

EMBRYONIC OR FETAL MEMBRANES

During the second week of development, with the embryo implanted in the uterus, cells within the
blastocyst start to organize into layers. Some grow to form the extra-embryonic membranes needed to
support and protect the growing embryo: the amnion, the yolk sac, the allantois, and the chorion (Figure
6).
1) Amnion. At the beginning of the second week, the cells of the inner cell mass form into a two-
layered disc of embryonic cells (it is composed of ectoderm inside and mesoderm outside), and a space.
The space between the embryo and the amnion is called the amniotic cavity which is filled with a clear,
watery fluid secreted by both the embryo and the membrane. The amniotic fluid prevents desiccation of
the embryo and acts as a protective cushion that absorbs shocks.
2) Yolk sac. On the ventral side of the embryonic disc, opposite the amnion, cells in the lower layer
of the embryonic disk (the hypoblast) extend into the blastocyst cavity and form a yolk sac. The yolk sac
supplies some nutrients absorbed from the trophoblast and also provides primitive blood circulation to the
developing embryo for the second and third week of development. When the placenta takes over
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nourishing the embryo at approximately week 4, the yolk sac has been greatly reduced in size and its main
function is to serve as the source of blood cells and germ cells (cells that will give rise to gametes).
3) Allantois. During week 3, a finger-like outpocketing of the yolk sac develops into the allantois (it
is composed of endoderm inside and the mesoderm outside), a primitive excretory duct of the embryo that
will become part of the urinary bladder. In human the allantois is small and nonfunctional except for
furnishing blood vessels to the placenta. Together, the stalks of the yolk sac and allantois establish the
outer structure of the umbilical cord.
4) Chorion. The last of the extra-embryonic membranes is the chorion, which is the one membrane
that surrounds all others. It is made up ectoderm outside and mesoderm inside. It completely surrounds
the embryo and protects it. It also takes part in the formation of placenta.

Figure 6. Diagram showing fetal membranes (schematic)

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 GASTRULATION /FORMATION OF GERM LAYERS

Formation of gastrula from the blastula is called gastrulation. Gastrulation is that phase of embryonic
development during which the cells of blastula (in frogs)/blastodermic vesicle (in mammals) move in small
masses or as sheet of cells to attain the final location. Such movements of cells are called morphogenetic
movement. Gastrulation results in the formation of three germ layers, in all triploblastic animals three germ
layers are ectoderm, mesoderm and endoderm.
The embryo, which takes the shape of an oval-shaped disc, forms an indentation called the primitive
streak along the dorsal surface of the epiblast. A node at the caudal or “tail” end of the primitive streak
emits growth factors that direct cells to multiply and migrate. Cells migrate toward and through the primitive
streak and then move laterally to create two new layers of cells. The first layer is the endoderm, a sheet of
cells that displaces the hypoblast and lies adjacent to the yolk sac. The second layer of cells fills in as the
middle layer, or mesoderm. The cells of the epiblast that remain (not having migrated through the primitive
streak) become the ectoderm (Figure 7).

Figure 7. Formation of mesoderm and ectoderm in embryo.

1) Formation of endoderm. The blastocyst (blastodermic vesicle) grows in size by absorbing food
from the uterus. Some cells detach from the inner cell mass to form endoderm in the blastocoel. Endoderm
differentiates into the primitive gut. The primitive gut differentiates into two portions at a later stage. One
portion forms the gut tract and the other portion communicates with the gut of the embryo and is called the yolk
sac.

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 2) Formation of embryonic disc. Just after the formation of endoderm, the remaining mass of cells
of the inner cell mass is called embryonic disc. The embryonic disc at this stage consists of cephalic margin,
embryonic disc proper and caudal margin.
3) Formation of Mesoderm. It starts after the formation of endoderm. Formation of mesoderm starts
from the caudal rapid proliferation of cells. The proliferated cells detach from the embryonic disc and form
mesoderm.
4) Formation of ectoderm. After the formation of mesoderm, the remaining cells of the embryonic
disc form a layer, the ectoderm. Thus, the three germ layers, ectoderm, mesoderm and endoderm are formed.

ORGANOGENESIS

The development of organ and organ-systems in the embryo is called organogenesis. This follows
gastrulation. The process of gastrulation forms the three germ layers of the embryo namely ectoderm, mesoderm
and endoderm. All the organ-systems of the multicellular animals are derived from these three germ layers.

FATE OF THREE GERM LAYERS

Following gastrulation of the embryo in the third week, embryonic cells of the ectoderm, mesoderm, and
endoderm begin to migrate and differentiate into the cell lineages that will give rise to mature organs and
organ systems in the infant.
Ectoderm. It forms 1. Epidermis of skin, epidermal derivatives like epidermal glands, hair, nails, etc.
2. Chromatophoies (pigment cells of the skin). 3. Entire nervous system. 4. Conjunctives cornea and lens
of eyes. 5. Iris and ciliary muscles of eyes. 6. Nasal and olfactory epithelia; 7. Outer layer of tympanic
membrane. 8. Internal ears. 9. Enamel of teeth. 10. Epithelium of stomodaeum (fore gut) and protoderm
(hind gut). II. Some glands—sweat glands, sebaceous glands, mammary glands, lacrimal glands, salivary
glands, pituitary gland, pineal gland and medulla of adrenal glands.
Mesoderm. It gives rise to 1. Dermis of skin. 2. Muscles except iris and ciliary muscles. 3. Connective
tissues. 4. Dentine of teeth. 5. Heart and blood vessels. 6. Blood and lymph. 7. Spleen. 8. Pericardium of
heart and pleura of lungs. 9. Sclera and choroid of eyes. 10. Connective tissue basis of tympanic membrane.
11. Cortex of the adrenal glands. 12. Notochord. 13. Coelomic epithelia. 14. Mesenteries. 15. Most of
skeleton. 16. Excretory system with the exception of the urinary bladder and urethra. 17. Reproductive
system with the exception of the prostate.
Endoderm. It forms. 1. Mucosa (mucous membrane) of the gut (alimentary canal) with the exception
of those portions already mentioned in the ectoderm. 2. Some glands — pancreas, liver, glands in the wall
of gastrointestinal tract (gastric glands, intestinal glands), thyroid, parathyroids, thymus and greater part
of prostate. 3. Inner layer of tympanic membrane. 4. The epithelial lining of the Eustachian tube and
tympanic cavity. 5. Trachea, bronchi and lungs. 6. Urinary bladder. 7. Urethra.
Thus, tympanic membrane is derived from all the three germ layers: ectoderm, mesoderm and
endoderm.

PLACENTA

The outer surface of the chorion in humans develops a number of fingers like projections, known as
chorionic villi, which grow into the tissue of the uterus. These villi, penetrate the tissues of the uterine wall
in which they are embedded, make up the organ known as the placenta, by means of which the developing
embryo obtains nutrients and oxygen and gets rid of carbon dioxide and metabolic wastes. There are many
capillaries in the villi that receive blood from the embryo by way of one of the two umbilical arteries and
return it to the embryo by way of the umbilical vein. The bloods of the mother and fetus do not mix at all
in the placenta or any other place. The blood of the fetus in the capillaries of the chronic villi comes in close
contact with the mother's blood in the tissues between the villi, but they are always separated by a

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membrane, through which substances must diffuse or be transported by some active, energy- requiring
process.

The placenta performs the following functions.

1) Nutrition. All the nutritive elements from the maternal blood pass into the fetus through the placenta.
2) Respiration. Oxygen passes from the maternal blood to the fetal blood through the placenta, and
carbon dioxide passes in the reverse direction.
3) Excretion. The fetal excretory products diffuse into the maternal blood through placenta and are
excreted by the mother.
4) Storage. Placenta stores glycogen, fat, etc.
5) As a Barrier. Placenta serves as an efficient barrier and allows those materials to pass into the
fetal blood that are necessary.
6) Endocrine Function. Placenta secretes hormones such as estrogens, progesterone and human
chorionic gonadotropin.

GLOSSARY:
Capacitation: The process by which the sperm becomes capable of fertilizing an egg.
Acrosome Reaction: A regulated exocytotic event in which an apical vesicle in the sperm head fuses
with the sperm plasma membrane. The acrosome reaction is triggered in response to egg factors.
Cortical Reaction: A regulated exocytosis in which apically localized vesicles (cortical granules) in the
egg fuse with plasma membrane after fertilization.
Zona Pellucida: A coat surrounding the egg that contains three glycoproteins.
Pronuclei: The transitional male and female nuclei formed in the egg after fertilization. They fuse to
form the diploid zygote nucleus.
Polyspermy: The condition in which more than one sperm fertilizes an egg. Polyspermy leads to
defective development.

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