BARR BODY /SEX CHROMATIN /X- CHROMATIN

Aim: To demonstrate the presence of Barr body from the oral mucosa of a human female

Requirements: Light microscope, glass slide, coverslip, flat toothpick, nailpaint, methylene blue
stain

Procedure:
1. Take a clean glass slide
2. Place a drop of water in the center.
3. Gently scrape the inside of cheek (of girl students of the class) with the help of the flat side of
the toothpick.
4. Spread the oral smear in water.
5. Put a drop of methylene blue stain (a vital dye)
6. Put a cover slip and seal the ends of the coverslip with a nail paint.
7. Observe under a microscope.
8. Focus on the squamous epithelial cells n then observe the nucleus of the cell under high power.
9. Repeat the same procedure with the oral smear of the boys of the class. This serves as the
control.

Observation:
One can observe a darkly stained body just below the nuclear membrane in about half the cells.
This is the Barr body which is not present in all cells but in about half the cells.
Discussion

In 1949, Barr and Bertram made an important discovery that the interphase nucleus of nerve
cells from female cats contains a small chromatin body which is lacking in males. Cell nuclei
from female cells contained a small, darkly stained body near the nuclear envelope, which
could be used for sexual dimorphism. This was called sex chromatin or the Barr body and after
the 1971 Paris conference, the X- chromatin.
It is now known that this difference is present in cell nuclei from most of the mammalian groups
including humans. This darkly stained body was named the Barr body to honor his work.
Barr body is found in cells of organisms where sex is determined by the presence of Y or W
chromosome.
Location:
X-chromatin can be found in different positions within the nucleus. For example, in nerve cells it
may be near the nucleolus, in the nucleoplasm, or near the nuclear envelope. In cells of the oral
mucosa, it is generally attached to the nuclear envelope. In neutrophils, it may appear as a small
rod called drumstick.
Frequency of occurrence:
In nervous tissue the frequency may be 85%, in whole mounts of amniotic or chorionic epithelium
it may be as high as 96%. In oral smears it may vary between 20 and 50% in normal females.
No. of Barr bodies:
Sex chromatin is derived from one of the X-Chromosomes. The other chromosome is euchromatic
and is decondensed at interphase. The no. of Barr bodies at interphase is therefore nX-1. This
means that the no. of Barr bodies is one less than the total no. of X chromosomes.
Lyon’s Hypothesis /Inactive X-Hypothesis / Dosage compensation
Mary. F. Lyon in 1972, gave this hypothesis according to which
1) Only one of the X- chromosome is genetically active
2) The X undergoing condensation may be either of maternal or paternal origin and the decision
by which X becomes inactive is random
3) The inactivation occurs early in embryonic life and remains fixed.
Dosage compensation:
In those cases where sex is determined by XY male mechanism, the females have two X
chromosomes while the males have just one .This means that the genes located on the X
chromosomes are present in double dose in females as compared to the males. Therefore, to
compensate for this, one of the X chromosome is inactive or heterochromatic in females and
remains so.
Practical Implications:
1) It offers the possibility of relating the origin of certain congenital diseases to chromosomal
anomalies included among these is the diagnosis of sex in intersexual states in post-natal and
in fetal life.
2) The no, of Barr bodies give an insight about the abnormal karyotypes of an individual. for
example, Turner’s syndrome (no sex chromatin), Klinefelter’s syndrome (1,2 and 3 depending
on the no. of X chromosomes )