Abstract
DNA methylation is an epigenetical mechanism that plays crucial roles in cellular differentiation and tissue development in embryogenesis. The aim of the present study was to determine the effects of a demethylating agent, 5-azacytidine, on testicular development during embryonal life in mouse. Ten pregnant mice were administered 5-azacytidine (5-azaC) (i.p. 2 mg/kg of agent dissolved in 0.1 mg/ml PBS) during 8th (Group 1), 11th (Group 2), 14th (Group 3) and 18th (Group 4) days of pregnancy periods and male siblings of these animals were obtained (experimental groups) whereas the control group animals received no treatment and siblings of this group were also obtained. Testicular tissues from all groups were taken 20 days after birth and examined at the light and electron microscopical levels. All pregnancies were terminated in Group 1 animals, therefore no observations could be done in this group. While Group 2 and 3 siblings showed distinctive kongenital abnormalities such as; anancephaly, growth failure, cleft palate, extremity abnormalities, supernumerary ribs and whirled shaped-tails, no such abnormalities were observed in Group 4 when compared to the control group. Microscopical examination of testicular tissues in groups 2 and 3 demonstrated cellular disintegration of spermatocytes in seminiferous tubules. In addition, cytoplasmic vacuoles and thickening of the basement membrane were also evident in both groups 2 and 3. Apoptotic-like cells were seen especially in group 2 and rarely in group 3. There were no structural alterations in group 4 animals, except a decreased number of spermatocytes in seminiferous tubules when compared to the control group, possibly indicating the completion of embryogenesis in this group. In conclusion, it could be suggested that the demethylating agent 5-azacytidine may trigger an unknown gene reactivation during early embryogenesis possibly affecting the cell and tissue differentiation in developing mammalian embryos.
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