Chromosomes

What Exactly is a
chromosome?
Chromosomes are the rod-shaped,
filamentous bodies present in the
nucleus, which become visible during
cell division.

They are the carriers of the gene or unit

of heredity.

Chromosome are not visible in active

nucleus due to their high water content,
but are clearly seen during cell division.
 Chromosomes were first described
by Strausberger in 1875.
 The term “Chromosome”, however
was first used by Waldeyer in 1888.
 They were given the name
chromosome (Chromo = colour;
Soma = body) due to their marked
affinity for basic dyes.
 Their number can be counted easily
only during mitotic metaphase.
 Chromosomes are composed of thin
chromatin threads called Chromatin
fibers.
 These fibers undergo folding, coiling and
supercoiling during prophase so that the
chromosomes become progressively
thicker and smaller.
 Therefore, chromosomes become
readily observable under light
microscope.
 At the end of cell division, on the other
hand, the fibers uncoil and extend as
fine chromatin threads, which are not
visible at light microscope
 Number of chromosomes
 Normally, all the individuals of a species have
the same number of chromosomes.
 Closely related species usually have similar
chromosome numbers.
 Presence of a whole sets of chromosomes is
called euploidy.
 It includes haploids, diploids, triploids,
tetraploids etc.
 Gametes normally contain only one set of
chromosome – this number is called Haploid
 Somatic cells usually contain two sets of
chromosome - 2n : Diploid
3n – triploid
4n – tetraploid
The condition in which the chromosomes sets
are present in a multiples of “n” is
Polyploidy
When a change in the chromosome number
does not involve entire sets of
chromosomes, but only a few of the
chromosomes - is Aneuploidy.
 Monosomics (2n-1)
 Trisomics (2n+1)
 Nullisomics (2n-2)
 Tetrasomics (2n+2)
Organism No.
chromosomes
 Human 46
 Chimpanzee 48
 Dog 78
 Horse 64
 Chicken 78
 Goldfish 94
 Fruit fly 8
 Mosquito 6
 Nematode 11(m), 12(f)
 Horsetail 216
 Sequoia 22
 Round worm 2
Organism No. chromosomes

 Onion 16
 Mold 16
 Carrot 20
 Tomato 24
 Tobacco 48
 Rice 24
 Maize 20
 Haploppus gracilis 4
 Crepis capillaris 6
 On the extreme, round worm shows
only two chromosomes, while the
other extreme is represented by
Protozoa having 300 or more
chromosomes.
 However, most organisms have
numbers between 12 to 50.
 3-8 in fungi
 From 8 – 16 in Angiosperms (Most
common number being 12).
 Chromosome Size
 In contrast to other cell organelles, the size of
chromosomes shows a remarkable variation
depending upon the stages of cell division.
 Interphase: chromosome are longest & thinnest
 Prophase: there is a progressive decrease in
their length accompanied with an increase in
thickness
 Anaphase: chromosomes are smallest.
 Metaphase: Chromosomes are the most easily
observed and studied during metaphase when
they are very thick, quite short and well spread
in the cell.

 Therefore, chromosomes measurements are

generally taken during mitotic metaphase.
The size of the chromosomes in mitotic phase of
animal and plants sp generally varies between
0.5 µ and 32 µ in length, and between 0.2 µ
and 3.0 µ in diameter.
The longest metaphase chromosomes found in
Trillium - 32 µ.
The giant chromosomes found in diptera and
they may be as long as 300 µ and up to 10 µ in
diameter.
In general, plants have longer chromosomes
than animal and species having lower
chromosome numbers have long chromosomes
than those having higher chromosome numbers
Among plants, dicots in general, have a higher
number of chromosome than monocots.
Chromosomes are longer in monocot than dicots.
 In order to understand chromosomes and their
function, we need to be able to discriminate
among different chromosomes.
 First, chromosomes differ greatly in size
 Between organisms the size difference can be
over 100-fold, while within a sp, some
chromosomes are often 10 times as large as
others.
 In a species Karyotype, a pictorial or
photographic representation of all the different
chromosomes in a cell of an individual,
chromosomes are usually ordered by size and
numbered from largest to smallest.
 Can distinguish chromosomes by “painting” –
using DNA hybridization + fluorescent probes –
during mitosis
 Karyotype: is the general morphology
of the somatic chromosome. Generally,
karyotypes represent by arranging in
the descending order of size keeping
their centromeres in a straight line.
 Idiotype: the karyotype of a species
may be represented diagrammatically,
showing all the morphological features
of the chromosome; such a diagram is
known as Idiotype.
 Chromosomes may differ in the position
of the Centromere, the place on the
chromosome where spindle fibers are
attached during cell division.
 In general, if the centromere is near the
middle, the chromosome is
metacentric
 If the centromere is toward one end, the
chromosome is acrocentric or
submetacentric
 If the centromere is very near the end,
the chromosome is telocentric.
 The centromere divides the
chromosome into two arms, so that, for
example, an acrocentric chromosome
has one short and one long arm,
 While, a metacentric chromosome has
arms of equal length.
 All house mouse chromosomes are
telocentric, while human chromosomes
include both metacentric and
acrocentric, but no telocentric.
 Autosomal pair Sex
chromosome
Diploid No. of No. of
X Y
(2n) metacentrics acrocentric or
telocentric
Cat 38 16 2 M M
Dog 78 0 38 M A
Pig 38 12 6 M M
Goat 60 0 29 A M
Sheep 54 3 23 A M
Cow 60 0 29 M M
Horse 64 13 18 M A

M – Metacentric; A – Acrocentric
 Euchromatin and Heterochromatin
 Chromosomes may be identified by regions that stain
in a particular manner when treated with various
chemicals.
 Several different chemical techniques are used to
identify certain chromosomal regions by staining
then so that they form chromosomal bands.
 For example, darker bands are generally found near
the centromeres or on the ends (telomeres) of the
chromosome, while other regions do not stain as
strongly.
 The position of the dark-staining are
heterochromatic region or heterochromatin.
 Light staining are euchromatic region or
euchromatin.
 Heterochromatin is classified into two
groups: (i) Constitutive and (ii)
Facultative.
 Constitutive heterochromatin remains
permanently in the heterochromatic
stage, i.e., it does not revert to the
euchromatic stage.
 In contrast, facultative heterochromatin
consists of euchromatin that takes on
the staining and compactness
characteristics of heterochromatin
during some phase of development.
 Satellite DNAs

When the DNA of a prokaryote, such as E.coli,

is isolated, fragmented and centrifuged to
equilibrium in a Cesium chloride (CsCl)
density gradient, the DNA usually forms a
single band in the gradient.
On the other hand, CsCl density-gradient
analysis of DNA from eukaryotes usually
reveals the presence of one large band of
DNA (usually called “Mainband” DNA) and
one to several small bands.
These small bands are referred to as “Satellite
DNAs”.
For e.g., in Drosophila virilis, contain three
distinct satellite DNAs.
Prokaryotic and
Eukaryotic
Chromosomes
 Not only the genomes of eukaryotes are
more complex than prokaryotes, but the
DNA of eukaryotic cell is also organized
differently from that of prokaryotic cells.
 The genomes of prokaryotes are
contained in single chromosomes,
which are usually circular DNA
molecules.
 In contrast, the genomes of eukaryotes
are composed of multiple
chromosomes, each containing a linear
molecular of DNA.
 Although the numbers and sizes of
chromosomes vary considerably between
different species, their basic structure is the
same in all eukaryotes
Organism Genome
Chromosome
Size (Mb)a numbera
Arabidopsis thaliana 70 5
Corn 5000 10
Onion 15,000 8
Lily 50,000 12
Fruit fly 165 4
Chicken 50,000 39
Mouse 1,200 20
Cow 3000 30
Human 3000 23
a – both genome size and chromosome numbers are for
haploid cells
 The DNA of eukaryotic cell is tightly bound to
small basic proteins (histones) that package
the DNA in an orderly way in the cell nucleus.
 This task is substantial (necessary), given the
DNA content of most eukaryotes
 For e.g., the total extended length of DNA in a
human cell is nearly 2 m, but this must be fit
into a nucleus with a diameter of only 5 to
10µm.

 Although DNA packaging is also a problem in

bacteria, the mechanism by which prokaryotic
DNA are packaged in the cell appears distinct
from that eukaryotes and is not well
understood.
 Prokaryotic chromosome
 The prokaryotes usually have
only one chromosome, and it
bears little morphological
resemblance to eukaryotic
chromosomes.
 Among prokaryotes there is
considerable variation in
genome length bearing genes.
 The genome length is smallest
in RNA viruses
 In this case, the organism is
provided with only a few genes
in its chromosome.
 The number of gene may be as
high as 150 in some larger
bacteriophage genome.
 In E.coli, about 3000 to 4000 genes are
organized into its one circular chromosome.
 The chromosome exists as a highly folded
and coiled structure dispersed throughout
the cell.
 The folded nature of chromosome is due to
the incorporation of RNA with DNA.
 There are about 50 loops in the chromosome
of E.coli.
 These loops are highly twisted or
supercoiled structure with about four
million nucleotide pairs.
 Its molecular weight is about 2.8 X109
 During replication of DNA, the coiling must
be relaxed.
 DNA gyrase is necessary for the unwinding
the coils.
 Bacterial Chromosome
 Single, circular DNA molecule
located in the nucleoid region of cell
Supercoiling
 Supercoiling

Most common type

of supercoiling

The helix twists

on itself; twists
to the right Helix twists on
itself in the opposite
direction; twists to
the left
 Mechanism of folding of a bacterial
chromosome

There are many supercoiled loops (~100 in E. coli) attached to a

central core. Each loop can be independently relaxed or condensed.

Topoisomerase enzyme – (Type I and II) that introduce or remove

supercoiling.
 Chromatin
 The complexes between eukaryotic DNA and
proteins are called Chromatin, which typically
contains about twice as much protein as DNA.
 The major proteins of chromatin are the
histones – small proteins containing a high
proportion of basic aminoacids (arginine and
lysine) that facilitate binding negatively charged
DNA molecule .
 There are 5 major types of histones: H1, H2A,
H2B, H3, and H4 – which are very similar
among different sp of eukaryotes.
 The histones are extremely abundant proteins in
eukaryotic cells.
 Their mass is approximately equal to that of the
 The major histone proteins:

Histone Mol. Wt No. of Percentage

Amino acidLys + Arg
H1 22,500 244 30.8
H2A 13,960 129 20.2
H2B 13,774 125 22.4
H3 15,273 135 22.9
H4 11,236 102 24.5
The DNA double helix is bound to proteins called
histones. The histones have positively charged
(basic) amino acids to bind the negatively
charged (acidic) DNA. Here is an SDS gel of
histone proteins, separated by size
 In addition, chromatin contains an
approximately equal mass of a wide variety of
non-histone chromosomal proteins.
 There are more than a thousand different types
of these proteins, which are involved in a range
of activities, including DNA replication and
gene expression.
 The DNA of prokaryotes is similarly associated
with proteins, some of which presumably
function as histones do, packing the DNA
within the bacterial cell.
 Histones, however are unique feature of
eukaryotic cells and are responsible for distinct
structural organization of eukaryotic chromatin
 The basic structural unit of chromatin, the
nucleosome, was described by Roger Kornberg in
1974.
 Two types of experiments led to Kornberg’s proposal
of the nucleosome model.
 First, partial digestion of chromatin with micrococcal
nuclease (an enzyme that degrades DNA) was found
to yield DNA fragments approximately 200 base
pairs long.
 In contrast, a similar digestion of naked DNA (not
associated with protein) yielded a continuous smear
randomly sized fragments.
 These results suggest that the binding of proteins to
DNA in chromatin protects the regions of DNA from
nuclease digestion, so that enzyme can
attack DNA only at sites separated by
 Electron microscopy revealed that
chromatin fibers have a beaded
appearance, with the beads spaced at
intervals of approximately 200 base
pairs.
 Thus, both nuclease digestion and the
electron microscopic studies suggest
that chromatin is composed of repeating
200 base pair unit, which were called
nucleosome.
individual nucleosomes = “beads
on a string”
 Detailed analysis of these nucleosome
core particles has shown that they
contain 146 base pairs of DNA wrapped
1.75 times around a histone core
consisting of two molecules each of H2A,
H2B, H3, and H4 (the core histones).
 One molecule of the fifth histone H1, is
bound to the DNA as it enters and exists
each nucleosome core particle.
 This forms a chromatin subunit known
as chromatosome, which consist of 166
base pairs of DNA wrapped around
histone core and held in place by H1 (a
linker histone)
 Centromeres and
Telomeres
 Centromeres and telomeres are two
essential features of all eukaryotic
chromosomes.
 Each provide a unique function i.e.,
absolutely necessary for the stability of the
chromosome.
 Centromeres are required for the
segregation of the centromere during
meiosis and mitosis.
 Teleomeres provide terminal stability to
the chromosome and ensure its survival
 Centromere
 The region where two sister chromatids of a
chromosome appear to be joined or “held together”
during mitatic metaphase is called Centromere
 When chromosomes are stained they typically show a
dark-stained region that is the centromere.
 Also termed as Primary constriction
 During mitosis, the centromere that is shared by the
sister chromatids must divide so that the chromatids
can migrate to opposite poles of the cell.
 On the other hand, during the first meiotic division
the centromere of sister chromatids must remain
intact
 whereas during meiosis II they must act as they do
during mitosis.
 Therefore the centromere is an important
component of chromosome structure and
 As a result, centromeres are the first
parts of chromosomes to be seen
moving towards the opposite poles
during anaphase.
 The remaining regions of
chromosomes lag behind and appear
as if they were being pulled by the
centromere.
 Kinetochore
 Within the centromere region, most
species have several locations where
spindle fibers attach, and these sites
consist of DNA as well as protein.

 The actual location where the attachment

occurs is called the kinetochore and is
composed of both DNA and protein.

 The DNA sequence within these regions

is called CEN DNA.
 Typically CEN DNA is about 120 base
pairs long and consists of several sub-
domains, CDE-I, CDE-II and CDE-
III.
 Mutations in the first two sub-domains
have no effect upon segregation,
 but a point mutation in the CDE-III
sub-domain completely eliminates the
ability of the centromere to function
during chromosome segregation.
 Therefore CDE-III must be actively
involved in the binding of the spindle
fibers to the centromere.
 The protein component of the
kinetochore is only now being
characterized.
 A complex of three proteins called
Cbf-III binds to normal CDE-III
regions but can not bind to a CDE-III
region with a point mutation that
prevents mitotic segregation.
 Telomere
 The two ends of a chromosome are known
as telomeres.
 It required for the replication and stability
of the chromosome.
 When telomeres are damaged or removed
due to chromosome breakage, the damaged
chromosome ends can readily fuse or unite
with broken ends of other chromosome.
 Thus it is generally accepted that structural
integrity and individuality of chromosomes
is maintained due to telomeres.
 McClintock noticed that if two
chromosomes were broken in a cell, the
end of one could attach to the other and
vice versa.
 What she never observed was the
attachment of the broken end to the end
of an unbroken chromosome.
 Thus the ends of broken chromosomes
are sticky, whereas the normal end is not
sticky, suggesting the ends of
chromosomes have unique features.
until recently, little was known about
molecular structure of telomeres. However,
during the last few years, telomeres have been
isolated and characterized from several sp.
Species Repeat Sequence
Arabidopsis TTTAGGG
Human TTAGGG
Oxytricha TTTTGGGG
Slime Mold TAGGG
Tetrahymena TTGGGG
Trypanosome TAGGG
 Tetrahymena -
 The telomeres of this protozoa organism.
organism end in the
sequence 5'-TTGGGG-3'.
 The telomerase adds a
series of 5'-TTGGGG-3'
repeats to the ends of the
lagging strand.
 A hairpin occurs when
unusual base pairs
between guanine residues
in the repeat form.
 Finally, the hairpin is
removed at the 5'-
TTGGGG-3' repeat.
RNA Primer - Short stretches of
 Thus the end of the ribonucleotides (RNA substrates)
chromosome is faithfully found on the lagging strand during
DNA replication. Helps initiate
 Staining and Banding
chromosome
Staining procedures have been developed in the
past two decades and these techniques help to
study the karyotype in plants and animals.
1. Feulgen Staining:
Cells are subjected to a mild hydrolysis in
1N HCl at 600C for 10 minutes.
This treatment produces a free aldehyde
group in deoxyribose molecules.
When Schiff’s reagent (basic fuschin
bleached with sulfurous acid) to give a deep
pink colour.
Ribose of RNA will not form an aldehyde
under these conditions, and the reaction is thus
specific for DNA
2. Q banding:
The Q bands are the fluorescent bands
observed after quinacrine mustard staining
and observation with UV light.
The distal ends of each chromatid are not
stained by this technique.
The Y chromosome become brightly
fluorescent both in the interphase and in
metaphase.
3. R banding:
The R bands (from reverse) are those located
in the zones that do not fluoresce with the
quinacrine mustard, that is they are between
the Q bands and can be visualized as green.
4. G banding:
The G bands (from Giemsa) have the
same location as Q bands and do not
require fluorescent microscopy.
Many techniques are available, each
involving some pretreatment of the
chromosomes.
In ASG (Acid-Saline-Giemsa) cells
are incubated in citric acid and NaCl for
one hour at 600C and are then treated with
the Giemsa stain.
5. C banding:
The C bands correspond to
constitutive heterochromatin.
The heterochromatin regions in a
chromosome distinctly differ in their
stainability from euchromatic region.
VARIATION IN STRUTURE OF
CHROMOSOME
 Chromosomal
Aberrations
 The somatic (2n) and gametic (n) chromosome
numbers of a species ordinarily remain
constant.
 This is due to the extremely precise mitotic
and meiotic cell division.
 Somatic cells of a diploid species contain two
copies of each chromosome, which are called
homologous chromosome.
 Their gametes, therefore contain only one
copy of each chromosome, that is they contain
one chromosome complement or genome.
 Each chromosome of a genome contains a
definite numbers and kinds of genes, which
are arranged in a definite sequence.
 Chromosomal
Aberrations
 Sometime due to mutation or
spontaneous (without any known
causal factors), variation in
chromosomal number or structure
do arise in nature. - Chromosomal
aberrations.
 Chromosomal aberration may be
grouped into two broad classes:
1. Structural and 2. Numerical
 Structural Chromosomal
Aberrations
 Chromosome structure variations result
from chromosome breakage.
 Broken chromosomes tend to re-join; if
there is more than one break, rejoining
occurs at random and not necessarily
with the correct ends.
 The result is structural changes in the
chromosomes.
 Chromosome breakage is caused by X-
rays, various chemicals, and can also
occur spontaneously.
 There are four common type of
structural aberrations:
1. Deletion or Deficiency
2. Duplication or Repeat
3. Inversion, and
4. Translocation.
 Consider a normal chromosome with genes
in alphabetical order: a b c d e f g h i

1. Deletion: part of the chromosome has

been removed: a b c g h i

2. Dupliction: part of the chromosome is

duplicated:
abcdefdefghi

3. Inversion: part of the chromosome has

been re-inserted in reverse order: a b c f e
dghi
ring: the ends of the chromosome
are joined together to make a ring
4. translocation: parts of two non-
homologous chromosomes are joined:
If one normal chromosome is a b c
d e f g h i and the other chromosome is
u v w x y z,
then a translocation between them
would be
a b c d e f x y z and u v w g h i.
 Deletion or deficiency
Loss of a chromosome segment is known as
deletion or deficiency
It can be terminal deletion or interstitial or
intercalary deletion.
A single break near the end of the chromosome
would be expected to result in terminal
deficiency.
If two breaks occur, a section may be deleted and
an intercalary deficiency created.
Terminal deficiencies might seem less complicated.
But majority of deficiencies detected are intercalary
type within the chromosome.
Deletion was the first structural aberration
detected by Bridges in 1917 from his genetic
studies on X chromosome of Drosophila.
 Deletion generally produce striking genetic
and physiological effects.
 When homozygous, most deletions are lethal,
because most genes are necessary for life and
a homozygous deletion would have zero
copies of some genes.
 When heterozygous, the genes on the normal
homologue are hemizygous: there is only 1
copy of those genes.
 Crossing over is absent in deleted region of a
chromosome since this region is present in
only one copy in deletion heterozygotes.
 In Drosophila, several deficiencies induced
the mutants like Blond, Pale, Beaded, Carved,
Notch, Minute etc.
Deletion in Prokaryotes:
Deletions are found in prokaryotes as
well, e.g., E.coli, T4 phage and Lambda phage.
In E.coli, deletions of up to 1 % of the bacterial
chromosome are known.
In lambda phage, however 20% of the genome
may be missing in some of the deletions.
Deletion in Human:
Chromosome deletions are usually lethal even
as heterozygotes, resulting in zygotic loss,
stillbirths, or infant death.
Sometimes, infants with small chromosome
deficiencies however, survive long enough to
permit the abnormal phenotype they express.
Cri-du-chat (Cat cry syndrome):
The name of the syndrome came from a catlike
mewing cry from small weak infants with the
disorder.
Other characteristics are microcephaly (small head),
broad face and saddle nose, physical and mental
retardation.
Cri-du-chat patients die in infancy or early childhood.
The chromosome deficiency is in the short arm of
chromosome 5 .
Myelocytic leukemia
Another human disorder that is associated with a
chromosome abnormality is chronic myelocytic
leukemia.
A deletion of chromosome 22 was described by
P.C.Nowell and Hungerford and was called
“Philadelphia” (Ph’) chromosome after the city in
 Duplication
The presence of an additional
chromosome segment, as compared to
that normally present in a nucleus is
known as Duplication.
 In a diploid organism, presence of a
chromosome segment in more than two
copies per nucleus is called duplication.
 Four types of duplication:

1. Tandem duplication
2. Reverse tandem duplication
3. Displaced duplication
4. Translocation duplication
 The extra chromosome segment may be
located immediately after the normal
segment in precisely the same orientation
forms the tandem
 When the gene sequence in the extra
segment of a tandem in the reverse order i.e,
inverted , it is known as reverse tandem
duplication
 In some cases, the extra segment may be
located in the same chromosome but away
from the normal segment – termed as
displaced duplication
 The additional chromosome segment is
located in a non-homologous chromosome is
 Origin
 Origin of duplication involves chromosome
breakage and reunion of chromosome
segment with its homologous chromosome.
 As a result, one of the two homologous
involved in the production of a duplication
ends up with a deficiency, while the other
has a duplication for the concerned segment.
 Another phenomenon, known as unequal
crossing over, also leads to exactly the same
consequences for small chromosome
segments.
 For e.g., duplication of the band 16A of X
chromosome of Drosophila produces Bar eye.
 This duplication is believed to originate due
to unequal crossing over between the two
normal X chromosomes of female.
 Inversion
 When a segment of chromosome is oriented in the
reverse direction, such segment said to be inverted and
the phenomenon is termed as inversion.
 The existence of inversion was first detected by
Strutevant and Plunkett in 1926.
 Inversion occur when parts of chromosomes become
detached , turn through 1800 and are reinserted in such
a way that the genes are in reversed order.
 For example, a certain segment may be broken in two
places, and the breaks may be in close proximity because
of chance loop in the chromosome.
 When they rejoin, the wrong ends may become
connected.
 The part on one side of the loop connects with broken
end different from the one with which it was formerly
connected.
 This leaves the other two broken ends to become
attached.
 The part within the loop thus becomes turned around or
 Inversion may be classified into two
types:
 Pericentric - include the centromere
 Paracentric - do not include the

centromere
 An inversion consists of two breaks in
one chromosome.
 The area between the breaks is
inverted (turned around), and then
reinserted and the breaks then unite
to the rest of the chromosome.
 If the inverted area includes the
centromere it is called a pericentric
inversion.
 If it does not, it is called a paracentric
inversion.
 Inversions in natural populations

 In natural populations, pericentric inversions

are much less frequent than paracentric
inversions.
 In many sp, however, pericentric inversions are
relatively common, e.g., in some grasshoppers.
 Paracentric inversions appear to be very
frequent in natural populations of Drosophila.
 Translocation
 Integration of a chromosome
segment into a nonhomologous
chromosome is known as
translocation.
 Three types:
1. simple translocation
2. shift
3. reciprocal translocation.
 Simple translocation: In this case,
terminal segment of a chromosome is
integrated at one end of a non-
homologous region. Simple
translocations are rather rare.
 Shift: In shift, an intercalary segment of
a chromosome is integrated within a
non-homologous chromosome. Such
translocations are known in the
populations of Drosophila, Neurospora
etc.
 Reciprocal translocation: It is
produced when two non-homologous
chromosomes exchange segments – i.e.,
segments reciprocally transferred.
 Non-Disjunction
 Generally during gametogenesis the
homologous chromosomes of each pair
separate out (disjunction) and are
equally distributed in the daughter cells.
 But sometime there is an unequal
distribution of chromosomes in the
daughter cells.
 The failure of separation of homologous
chromosome is called non-disjunction.
 This can occur either during mitosis or
meiosis or embryogenesis.
 Mitotic non-disjunction: The failure of
separation of homologous chromosomes
during mitosis is called mitotic non-
disjunction.
 It occurs after fertilization.
 May happen during first or second cleavage.
 Here, one blastomere will receive 45
chromosomes, while other will receive 47.
 Meiotic non-disjunction: The failure of
separation of homologous chromosomes
during meiosis is called mitotic non-
disjunction
 Occurs during gametogensis
 Here, one type contain 22 chromosome,
while other will be 24.
 Variation in chromosome
number
 Organism with one complete set of
chromosomes is said to be euploid
(applies to haploid and diploid
organisms).

 Aneuploidy - variation in the number of

individual chromosomes (but not the
total number of sets of chromosomes).

 The discovery of aneuploidy dates back

to 1916 when Bridges discovered XO
male and XXY female Drosophila, which
had 7 and 9 chromosomes respectively,
 More about
 Nullisomy - loss of oneAneuploidy
homologous
chromosome
pair. (e.g., Oat )
 Monosomy – loss of a
single
chromosome
(Maize).
 Trisomy - one extra
chromosome. (Datura)
 Tetrasomy - one extra
chromosome pair.
 Uses of Aneuploidy
 They have been used to determine the
phenotypic effect of loss or gain of different
chromosome
 Used to produce chromosome substitution
lines. Such lines yield information on the
effects of different chromosomes of a variety
in the same genetic background.
 They are also used to produce alien
addition and alien substitution lines.
These are useful in gene transfer from one
species to another.
 Aneuploidy permits the location of a gene as
well as of a linkage group onto a specific
chromosome.
 Trisomy in Humans
Down Syndrome
 The best known and most common chromosome
related syndrome.
 Formerly known as “Mongolism”
 1866, when a physician named John Langdon
Down published an essay in England in which he
described a set of children with common
features who were distinct from other children
with mental retardation he referred to as
“Mongoloids.”
 One child in every 800-1000 births has Down
syndrome
 250,000 in US has Down syndrome.
 The cost and maintaining Down syndrome case
 Patients having Down syndrome will Short
in stature (four feet tall) and had an
epicanthal fold, broad short skulls, wild
nostrils, large tongue, stubby hands
 Some babies may have short necks, small
hands, and short fingers.
 They are characterized as low in mentality.
 Down syndrome results if the extra
chromosome is number 21.
Amniocentesis for Detecting Aneuploidy
 Chromosomal abnormalities are
sufficiently well understood to permit
genetic counseling.
 A fetus may be checked in early stages of
development by karyotyping the cultured
cells obtained by a process called
amniocentesis.
 A sample of fluid will taken from mother
and fetal cells are cultured and after a
period of two to three weeks,
chromosomes in dividing cells can be
stained and observed.
 If three No.21 chromosomes are present,
Down syndrome confirmed.
 The risk for mothers less than 25
years of age to have the trisomy is
about 1 in 1500 births.
 At 40 years of age, 1 in 100 births
 At 45 years 1 in 40 births.
 Other Syndromes
Chromosome Nomenclature: 47,
+13
Chromosome formula: 2n+1
Clinical Syndrome: Trisomy-13
Estimated Frequency Birth:
1/20,000
Main Phenotypic Characteristics:
Mental deficiency and deafness,
minor muscle seizures, cleft lip,
cardiac anomalies
 Other Syndromes
Chromosome Nomenclature: 47, +18
Chromosome formula: 2n+1
Clinical Syndrome: Trisomy-18
Estimated Frequency Birth: 1/8,000
Main Phenotypic Characteristics:
Multiple congenital malformation
of many organs, malformed ears, small
mouth and nose with general elfin
appearance.
90% die in the first 6 months.
 Other Syndromes
Chromosome Nomenclature: 45, X
Chromosome formula: 2n - 1
Clinical Syndrome: Turner
Estimated Frequency Birth: 1/2,500 female
Main Phenotypic Characteristics:
Female with retarded sexual
development, usually sterile, short
stature, webbing of skin in neck region,
cardiovascular abnormalities, hearing
impairment.
 Other Syndromes
Chromosome Nomenclature: 47, XXY, 48,
XXXY,
48,XXYY, 49, XXXXY, 50,
XXXXXY
Chromosome formula: 2n+1; 2n+2; 2n+2;
2n+3; 2n+4
Clinical Syndrome: Klinefelter
Estimated Frequency Birth: 1/500 male borth
Main Phenotypic Characteristics:
Pitched voice, Male, subfertile with small
testes, developed breasts, feminine, long
limbs.
 Giant
chromosomes
 Found in certain
tissues e.g., salivary
glands of larvae, gut
epithelium, Malphigian
tubules and some fat
bodies, of some
Diptera (Drosophila,
Sciara, Rhyncosciara)

 These chromosomes
are very long and thick
(upto 200 times their
size during mitotic
metaphase in the case
of Drosophila)

 Hence they are known

 They are first discovered by Balbiani in
1881 in dipteran salivary glands and thus
also known as salivary gland chromosomes.

 But their significance was realized only

after the extensive studies by Painter
during 1930’s.

 Giant chromosomes have also been

discovered in suspensors of young embryos
of many plants, but these do not show the
bands so typical of salivary gland
chromosomes.
 He described the morphology in detail
and discovered the relation between
salivary gland chromosomes and germ
cell chromosomes.

 Slides of Drosophila giant

chromosomes are prepared by
squashing in acetocarmine the
salivary glands dissected out from the
larvae.

 The total length of D.melanogater

giant chromosomes is about 2,000µ.
 Giant chromosomes are made
up of several dark staining
regions called “bands”.
 It can be separated by
relatively light or non-staining
“interband” regions.
 The bands in Drosophila giant
chromosome are visible even
without staining, but after
staining they become very
sharp and clear.
 In Drosophila about 5000
bands can be recognized.
 Some of these bands are as
thick as 0.5µ, while some
may be only 0.05µ thick.

 About 25,000 base-pairs are

now estimated for each
band.

 All the available evidence

clearly shows that each giant
chromosome is composed of
numerous strands, each
strand representing one
chromatid.

 Therefore, these
chromosomes are also
known as “Polytene
chromosome”, and the
condition is referred to as
 The numerous strands of these
chromosomes are produced due to repeated
replication of the paired chromosomes
without any nuclear or cell division.
 So that the number of strands (chromatids)
in a chromosome doubles after every round
of DNA replication
 It is estimated that giant chromosomes of
Drosophila have about 1,024 strands
 In the case of Chironomous may have about
4,096 strands.
 The bands of giant chromosomes are
formed as a result of stacking over one
another of the chromomeres of all strands
present in them.
 Since chromatin fibers are highly
coiled in chromosomes, they stain
deeply.
 On the other hand, the chromatin
fibers in the interband regions are
fully extended, as a result these
regions take up very light stain.
 In Drosophila the location of many
genes is correlated with specific
bands in the connected chromosomes.
 In interband region do not have
atleast functional genes
 During certain stages of development,
specific bands and inter band regions are
associated with them greatly increase in
diameter and produced a structure called
Puffs or Balbiani rings.
 Puffs are believed to be produced due to
uncoiling of chromatin fibers present in
the concerned chromomeres.
 The puffs are sites of active RNA
synthesis.
Figure 3. Polytene chromosome map of Anopheles gambiae
 Lampbrush Chromosome
 It was given this name because it is similar in
appearance to the brushes used to clean lamp
chimneys in centuries past.
 First observed by Flemming in 1882.
 The name lampbrush was given by Ruckert in
1892.
 These are found in oocytic nuclei of
vertebrates (sharks, amphibians, reptiles and
birds)as well as in invertebrates (Sagitta,
sepia, Ehinaster and several species of
insects).
 Also found in plants – but most experiments in
oocytes.
 Lampbrush chromosomes are up to 800
µm long; thus they provide very favorable
material for cytological studies.
 The homologous chromosomes are paired
and each has duplicated to produce two
chromatids at the lampbrush stage.
 Each lampbrush chromosome contains a
central axial region, where the two
chromatids are highly condensed
 Each chromosome has several
chromomeres distributed over its length.
 From each chromomere, a pair of loops
emerges in the opposite directions vertical
 One loop represent
one chromatid, i.e.,
one DNA molecule.
 The size of the loop
may be ranging the
average of 9.5 µm
to about 200 µm
 The pairs of loops
are produced due
to uncoiling of the
two chromatin
fibers present in a
highly coiled state
in the
chromomeres.
 One end of each loop is thinner (thin
end) than the other end (thick end).
 There is extensive RNA synthesis at
the thin end of the loops, while there
is little or no RNA synthesis at the
thick end.
Phase-contrast and fluorescent
micrographs of
lampbrush chromosomes
 Dosage Compensation
 Sex Chromosomes: females XX,
males XY
 Females have two copies of every X-
linked gene; males have only one.
 How is this difference in gene dosage
compensated for? OR
 How to create equal amount of X
chromosome gene products in males
and females?
 Levels of enzymes or proteins
encoded by genes on the X
chromosome are the same in
both males and females

 Even though males have 1 X

chromosome and females have 2.
 G6PD, glucose 6 phosphate
dehydrogenase, gene is carried on the
X chromosome
 This gene codes for an enzyme that
breaks down sugar
 Females produce the same amount of
G6PD enzyme as males
 XXY and XXX individuals produce the
same about of G6PD as anyone else
 In cells with more than two X chromosomes,
only one X remains genetically active and all
the others become inactivated.
 In some cells the paternal allele is expressed
 In other cells the maternal allele is
expressed
 In XXX and XXXX females and XXY males
only 1 X is activated in any given cell the
rest are inactivated
 Barr Bodies
 1940’s two Canadian scientists
noticed a dark staining mass in
the nuclei of cat brain cells
 Found these dark staining spots
in female but not males
 This held for cats and humans
 They thought the spot was a
tightly condensed X chromosome
 Barr Bodies

Barr bodies represent the inactive X

chromosome and are normally found
only in female somatic cells.
A woman with
the
chromosome
constitution
47, XXX
should have 2
Barr bodies in
each cell.
XXY

individuals are
male, but have
a Barr body.
 XO
individuals are
 Which chromosome is inactive is a matter
of chance, but once an X has become
inactivated , all cells arising from that
cell will keep the same inactive X
chromosome.
 In the mouse, the inactivation apparently
occurs in early in development
 In human embryos, sex chromatin bodies
have been observed by the 16th day of
gestation.
Mechanism of X-chromosome
Inactivation
 A region of the p arm of the X
chromosome near the centromere called
the X-inactivation center (XIC) is the
control unit.
 This region contains the gene for X-
inactive specific transcript (XIST). This
RNA presumably coats the X
chromosome that expresses it and then
DNA methylation locks the chromosome
in the inactive state.
 This occurs about 16 days after
fertilization in a female embryo.
 The process is independent from cell
to cell.
 A maternal or paternal X is randomly
chosen to be inactivated.
 Rollin Hotchkiss first discovered
methylated DNA in 1948.
 He found that DNA from certain sources
contained, in addition to the standard four
bases, a fifth: 5-methyl cytosine.
 It took almost three decades to find a role
for it.
 In the mid-1970s, Harold Weintraub and
his colleagues noticed that active genes are
low in methyl groups or under methylated.
 Therefore, a relationship between under
methylation and gene activity seemed
likely, as if methylation helped repress
genes.
 This would be a valuable means of
keeping genes inactive if methylation
passed on from parent to daughter cells
during cell division.
 Each parental strand retains its methyl
groups, which serve as signals to the
methylating apparatus to place methyl
groups on the newly made progeny
strand.
 Thus methylation has two of the
requirements for mechanism of
determination:
 1. It represses gene activity
 Strictly speaking, the DNA is altered,
since methyl groups are attached, but
because methyl cytosine behaves the
same as ordinary cytosine, the genetic
coding remain same.
 A striking example of such a role of
methylation is seen in the inactivation of
the X chromosome in female mammal.
 The inactive X chromosome become
heterochromatic and appears as a dark
fleck under the microscope – this
chromosome said to be lyonized, in
honor of Mary Lyon who first postulated
the effect in mice.
 An obvious explanation is that the DNA in the
lyonized X chromosome is methylated, where
as the DNA in the active, X chromosome is
not.
 To check this hypothesis Peter Jones and
Lawrence Shapiro grew cells in the presence
of drug 5-azacytosine, which prevents DNA
methylation.
 This reactivated the lyonized the X
chromosome.
 Furthermore, Shapiro showed these
reactivated chromosomes could be
transferred to other cells and still remain
active.