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In those species (including humans) in which sex is determined by the presence of the Y or W chromosome. a Barr body is the inactive X chromosome in a female cell, or the inactive Z in a male (Lyon, 2003). The process of inactivation is called Lyonization. The Lyon hypothesis states that in cells with multiple X chromosomes, all but one are inactivated during mammalian embryogenesis (Lyon, 1961). This happens early in embryonic development at random in mammals, (Brown, 1997) except in marsupials and in some extra-embryonic tissues of some placental mammals, in which the father's X chromosome is always deactivated (Lee, 2003). Barr bodies are named after their discoverer, Murray Barr (Barr & Bertram, 1949). In men and women with more than one X chromosome, the number of Barr bodies visible at interphase is always one less than the total number of X chromosomes normal female ( XX) having one Barr body in the nucleus of each undividing cell , the normal male (XY) will not have any Barr body but the male with
will have one Barr body in each cell. This technique is used in cytogenetic studies for the diagnosis of genetic disorders like Klinefelter’s syndrome, super female (XXX), etc.,
A Barr Body can be defined as a heterochromatic inactive X chromosome found in the nucleus of an undividing cell. Since random chromosomes are selected for inactivation early in embryonic development, this results in different regions of the adult body having different chromosomes inactivated. The Barr body chromosome is generally considered to be inert, but in fact a small number of genes remain active and expressed in some species.
Mechanism of Barr body formation
Mammalian X-chromosome inactivation is initiated from the X inactivation centre or “Xic” centre, usually found near the centromere. The center contains twelve genes, seven of which code for proteins, five for untranslated RNAs, of which only two are known to play an active role in the X inactivation process, Xist and Tsix. The centre also appears to be important in chromosome counting: ensuring that random inactivation only takes place when two X-chromosomes are present.
In 1949 Barr and Bertram reported that nerve cells from cats showed a sexual difference (dimorphism). Cell nuclei from female cells consistently showed a small darkly staining body near the nuclear envelope, while cell nuclei from males showed no such structure. It is now known that this difference is present in most cell nuclei from most of the mammalian groups, including humans. The darkly staining structure found only in the nuclei of cells from females is known as the Barr body. Further research has shown that the Barr body is actually an inactivated X chromosome. For proper functioning, most cell types must have one and only one active X chromosome at any given time. Since each female cell has two X chromosomes, one of them must be inactivated for normal cell functioning. Male cells have only one X, so they don't have a spare to inactivate and display as a Barr body. Infrequently, some females end up with three X chromosomes (XXX or trisomy X). The cell nuclei of these individuals show two Barr bodies. The fact that the number of X chromosomes is directly related to the number of Barr bodies in the nucleus is commonly used to confirm the sex of women athletes.
In this section, you will make a cheek cell smear as above, but use a sensitive DNA stain (Thionin) that will make Barr bodies visible.
1. Wash your slide in detergent and rinse well in deionized water. Rinse the slide in a stream of 70% ethyl alcohol from a squeeze bottle. Allow the slide to air dry on a clean paper towel. 2. To avoid contamination with bacteria, which stain nicely and might look like a Barr body, rinse your mouth very well with tap water at least three times. 3. Immediately after rinsing, Gently scrape the inside of your cheek with the toothpick and then smear it across the slide. 4. Allow the slide to air dry for 1 minute.