Correlation between the granulosa cell layer and active caspase-3 expression in ovarian follicles of Tropidurus hispidus and T. semitaeniatus (Squamata, Tropiduridae): immunohistochemical approach

Abstract

The greatest threats to terrestrial reptiles are urban development and habitat modification. In this sense, a better understanding of folliculogenesis in these animals would be important to knowledge of reproductive biology. The aim of this study was to analyze the correlation between the thickness of the granulosa cell layer and the expression of the active caspase-3 protein in the previtellogenic and vitellogenic follicles of T. hispidus and T. semitaeniatus . Ovaries were used for histological (morphology and morphometry: thickness of granulosa layer) and immunohistochemical (active caspase-3 expression) analyses. The previtellogenic follicles of T. hispidus and T. semitaeniatus showed a thicker granulosa layer, with pyriform and small cells. The vitellogenic follicles had a monolayer of cuboid cells, and a thicker thecal layer. The thickness of the granulosa layer was significantly higher in the previtellogenic compared to the vitellogenic phase for both species. However, no differences were observed between the species. Active caspase-3 was observed in the pyriform and intermediate cells in previtellogenesis of T. hispidus and T. semitaeniatus . Nevertheless, no immunostaining was observed in the vitellogenic phase in both species. In conclusion, this study shows that the thickness of the granulosa cell layer is higher in the previtellogenic follicles compared to the vitellogenic follicles in the two Tropidurus species. Pyriform and intermediate cells from previtellogenic follicles show high expression of the protein, indicating that remodeling of the epithelium is associated with apoptosis. Finally, our results provide a scientific basis for assisted reproductive techniques and conservation actions to the reptiles in the future.