UC Davis

The use of the RNA interference with its transient knockdown of genes offers a versatile technique to study the roles of FSH signaling on the development and function of ovarian follicles. The objective of this study was to develop a reliable protocol to knock down the follicle-stimulating hormone receptor in bovine granulosa cells utilizing the RNA interference pathway with small interfering RNAs (siRNAs). This work was done in bovine granulosa cells to understand further follicle-stimulation stimulating hormone signaling mediated by its receptor. FSH signaling is responsible for activating pro-apoptotic, mitogenic, and steroidogenic signals that characterize the development and differentiation of granulosa cells during folliculogenesis. This study tested several approaches to find the optimal conditions to knock down the FSHR in granulosa cells using siRNA. These approaches include testing the duration of transfection, different formulations and concentrations of the transfection reagent, the number of siRNA sequences used, and the siRNA concentration. Experiments are still ongoing to establish the optimal conditions for the knockdown of the FSHR in bovine granulosa cells.