Adaptive and Biological Responses of Buffalo Granulosa Cells Exposed to Heat Stress under In Vitro Condition

SIMPLE SUMMARY: The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to the control and other heat-treated groups. These results will provide insights into ways that tropically adapted breeds may be able to maintain better reproductive function when exposed to heat stress compared to temperate breeds. ABSTRACT: The steroidogenesis capacity and adaptive response of follicular granulosa cells (GCs) to heat stress were assessed together with the underlying regulating molecular mechanisms in Egyptian buffalo. In vitro cultured GCs were exposed to heat stress treatments at 39.5, 40.5, or 41.5 °C for the final 24 h of the culture period (7 days), while the control group was kept under normal conditions (37 °C). Comparable viability was observed between the control and heat-treated GCs at 39.5 and 40.5 °C. A higher release of E2, P4 and IGF-1 was observed in the 40.5 °C group compared with the 39.5 or 41.5 °C groups. The total antioxidant capacity was higher in response to heat stress at 39.5 °C. At 40.5 °C, a significant upregulation pattern was found in the expression of the stress resistance transcripts (SOD2 and NFE2L2) and of CPT2. The relative abundance of ATP5F1A was significantly downregulated for all heat-treated groups compared to the control, while TNFα was downregulated in GCs at 39.5 °C. Expression analyses of stress-related miRNAs (miR-1246, miR-181a and miR-27b) exhibited a significant downregulation in the 40.5 °C group compared to the control, whereas miR-708 was upregulated in the 39.5 and 40.5 °C groups. In conclusion, buffalo GCs exhibited different adaptive responses, to the different heat stress conditions. The integration mechanism between the molecular and secretory actions of the GCs cultured at 40.5 °C might provide possible insights into the biological mechanism through which buffalo GCs react to heat stress.