Evaluation of immunohistochemical markers of germ cells’ proliferation in the developing rat testis: A comparative study

Germ cells’ proliferation during testicular organogenesis in Wistar rat embryos and neonates [14.5, 18.5, 20.5 days post conception (dpc), birth (day 0), 1, 3, 5, 7 days post partum (dpp)] was evaluated via immunohistochemistry, using the PCNA and Ki-67 nuclear antibodies. Estimation of the reactive/total cell ratio, per visual field [labeIing index (LI)] was achieved using the Image Pro Plus Software. Immunostaining of the fetal testis, with both antibodies, revealed increasing germ cells’ numbers between 14.5 dpc and birth. From birth onwards, a sharp decline of germ cells’ population was observed in the first 3 days of postnatal life. Then, a transient increase of the LI, between 3 and 5 dpp, was noted. Afterwards, proliferation of germ cells ceased. These results indicate that, during fetal and neonatal life, two peaks of proliferative activity of germ cells are noticed. Following estimation of the LI for both PCNA and Ki-67, a prominent labeling for the first antibody was observed throughout the examined period. Ki-67 staining follows a similar pattern, showing, however, significant fluctuation in the obtained values, in comparison to PCNA. The significant differences observed don’t seem to be simply a result of the different half lives of the two markers, but rather a consequence of additional underlying cellular activity associated with PCNA, such as DNA repair.