Effects of estradiol on bovine thecal cell function in vitro: dependence on insulin and gonadotropins

The objective of this study was to evaluate the influence of estradiol (E2) on proliferation and steroid production by thecal cells obtained from large (≥8 mm) follicles of cattle. Five experiments evaluated the effect of various doses of E2 during a 2-d exposure in serum-free medium on hormone-induced steroidogenesis and cell proliferation. In LH-treated thecal cells of experiment 1, 300 ng/mL of E2 decreased progesterone production by 30% and increased androstenedione production to 5.8-fold of controls. In the absence of LH, both 3 and 300 ng/mL of E2 increased progesterone production. In experiment 2, in the presence of insulin and LH, 3, 30, and 300 ng/mL of E2 decreased progesterone production (by 17 to 36%), whereas 3 ng/mL of E2 decreased and 300 ng/mL of E2 increased androstenedione production. Doses of LH (3 to 30 ng/mL) tested in experiment 3 increased (to as much as 3.7-fold) progesterone production by thecal cells and E2 attenuated this stimulatory effect by 40%. In contrast, E2 amplified the stimulatory effect of LH on androstenedione production in experiment 3. In experiment 4, E2 (300 ng/mL) decreased IGF-I- and insulin-induced thecal cell progesterone production by 70 to 77%, whereas E2 increased basal, IGF-I, and insulin-induced androstenedione production. In experiment 5, in the presence of insulin, 10 to 1000 ng/mL of E2 had no effect on ¹²⁵I]-IGF-I binding to thecal cells, whereas 10 and 100 ng/mL of E2 increased and 1000 ng/mL of E2 decreased progesterone production by thecal cells. Estradiol had no consistent effect on thecal cell numbers among the 5 experiments. These results support the hypothesis that E2 may act as a paracrine factor to directly regulate hormone-induced steroid production by thecal cells without affecting cell numbers or numbers of insulin-like growth factor type I receptors.