Chromosomal Banding in Fishes

Dr.J.P.Srivastava (Assistant professor)

Department of Zoology
D.B.S.College
CSJM UNIVERSITY
KANPUR
 Meaning of Chromosomal Band:
A band is defined as that part of a chromosome, which clearly
distinguished from its adjacent segment by appearing light
(bright) and dark stripes or bands, which appear along its
length after being stained with specific dyes.

Figure 1: Characteristic banding pattern produced by exposure to trypsin during G banding. G

band light regions tend to be gene rich. G band dark regions tend to be gene poor. Giemsa
banding is a technique used in cytogenetics to produce a visible karyotype by staining condensed
chromosomes.
The stained chromosomes
when visualized under
microscope show a
continuous series of bright
and dark bands (or fluorescent
versus non-fluorescent).

Importantly, each chromosome

displays a unique banding
pattern, analogous to
“barcode”, which allows it to
be reliably differentiated from
other chromo­somes of the
same size and centromeric
position (fig 36.1).
The causes of many
diseases in human
beings can be identified
now on the basis of
molecular genetics.
Wolf Parkinson
Syndrome is caused
due to mutation in 7q3,
which means that the
defect is due to
chromosome 7 and in
the q arm at band
three(fig-36.2).
 Structure of Chromosome Band:
To understand what chromosomal bands represent, it is essential to know
structure of chromosome.
Eukaryotic chromosomes are composed of chromatin, a combination of
nuclear DNA and proteins.
There are two varieties of chromatin, one is known as heterochromatin and
another is called as euchromatin.
• Heterochromatin is believed to serve several functions,
from gene regulation to the protection of the integrity of
chromosome.
• The constitutive heterochromatin occurs around the
chromosome centromere and near telomeres.
• All cells of a given species will package the same region of
DNA in constitutive heterochromatin, and in all cells any
genes contained within the constitutive heterochromatin will
be poorly expressed.
• The facultative heterochromatin will not be consistent within
the cells of a species, and thus sequences in one cell that
is packed in facultative heterochromatin (and the genes
within poorly expressed) may be packed in euchromatin in
another cell (and genes within no longer silenced).
• Hence, the formation of facultative heterochromatin is
regulated, and is often associated with morphogenesis or
differentiation.
The two types of proteins are histones
and non-histones proteins.
Histones are proteins rich in positively
charged amino acids, lysine and
arginine.
For this reason they bind tightly to
negatively charged phosphates in DNA.
The non-histone proteins are mostly
transcription factors that regulate that
part of DNA which transcribe into RNA.
The banding techniques falls in two groups:
1. G-Q and R-bands, these bands are distributed along the length
of the whole chromosome.
2. C-bands (centromeric bands) and NOR’s (nucleolus organizer
regions). They are used to stain a restriction number of specific
bands or structures.
C-banding methods do not permit identification of every
chromosome in the somatic cell complement but can be used to
identify specific chromosomes.
G Banding:
• G bands are obtained by stain with Giemsa (hence called G-
bands) after digesting the chromosomes with trypsin or by
acetic-saline.
• Trypsin demonstrated a conspicuous banding pattern in almost
all the chromosomes of the complement.
• In the G-band, dark region contains heterochromatin, which is
late replicating and is rich in adenine and thymine (AT).
• The centromeres for the most part stained weakly.
• That is, they were negative for G-banding, showing that these
regions are sensitive to the proteolytic action of trypsin, while
most of the telomeres, which are heterochromatic, displayed
strong staining, and were therefore not digested by trypsin.
• The B micro-ch­romosome could not be visualized in the G-
banding preparations.
• In fish, I. labrosus, G-banding was prominent in almost all the
diploid number of 56 chromosomes as noticed by de
Carvalhoc and Dias (2005). However, centromeres showing
negative G-banding.
• In a study of the family Pimelodiade in (2005) utilized G-
banding in chromosome preparations of Steindachneridion
scripta and Pseudoplatystoms corruscans and, found a
pattern of longitudinal chromosomal differentiation in the
three species.
• These authors also reported the occurrence of C-banding in
the heterochromatin of the telomeric regions in most of the
chromosomes of I. labrosus from the Capivara Reservoir at
the two localities, few chromosomes showed C-banding
positive centromeres.
• When present, the supernumerary or B micro-chromosome
appeared totally heterochromatic.
• G-bands were also observed in Indian fishes, such as Channa
punctatus, Colisa fascieatus, Mystus tengara, Puntius
sophore and Labeo rohita.
• R-bands are approximately the reverse of G-bands (the R
stands for “reverse”). The dark regions are euchromatin and
where as bright regions are heterochromatin. R-banding is
obtained by heat and use of Giemsa or fluorescence.
Florescence G- R-bands are the photographic negatives of
the bright field versions i.e. the reverse of the bright field G-
band and R-bands.
• C Banding:
C-banding is done by de-purification with acid,
denaturing is carried out by base and extraction of
non-heterochromatic DNA in hot salt solutions as
stated by Coming (1978) then stained with Giesma
stain.
It stains areas of constitutive heterochromatin, which
is tightly packed and contains repetitive DNA.
Q Banding:
In this technique the chromosomes are stained by
fluorescent quinacrine dye and the chromosomes
show intense fluorescent (quinacrine) Q-bands.
These bands are rich in adenine and thymine (AT).
NOR Silver-Staining:
• This procedure help in identifying genes for ribosomal
RNA in a previous cell cycle.
• NOR banding is carried out with the help of silver
staining with fluorochrome chromomycin A3 (CMA)
distinguishing chromosomal sites of 18s ribosomal RNA.
The region is rich in Guanine and Cytosine (CG).
• If stain with mithraycin, it apparently stain DNA.
• It has been studied in about 200 species of fishes.
• In Cyprinus carpio one pair of NOR was noticed while
interstitial NOR was reported in Channa punctatus by
Rishi (1972).
• Recently, Swarca (2005) observed secondary
constrictions on the short arm of the first pair of
acrocentrics, shown to be associated with molecular
organizer region in Zungaro zungaro (Pisces,
Pimelodidae).
Figure 2. Metacentric (left), submetacentric (centre) and acrocentric (right) chromosomes.