Insulin-like growth factor-II stimulates steroidogenesis in cultured bovine thecal cells

Abstract

The objective of the present study was to determine the effects of insulin-like growth factor-II (IGF-II) on luteinizing hormone (LH)-induced progesterone and androstenedione production by bovine thecal cells and compare it to that of insulin and IGF-I. Cells from large (>7.9 mm) bovine follicles were collected and cultured for 2 days in the presence of 10% fetal calf serum. Then cells were cultured for an additional 1 or 2 days in serum-free medium with various doses of recombinant human IGF-II, bovine LH (30 ng/ml), IGF-I, and(or) insulin. Cell numbers were determined at the end of treatments via Coulter counting and used to correct steroid production data. In the presence of LH, 1-day treatment with 3–300 ng/ml of IGF-II had no significant effect on progesterone or androstenedione production, whereas 2-day treatment with 30, 100 and 300 ng/ml of IGF-II increased ( P < 0.05) both progesterone and androstenedione production by 2–3-fold. The estimated effective dose of IGF-II stimulating 50% of the maximal steroidogenic response was calculated to be 25 ng/ml. In the absence of LH, 2-day treatment of IGF-I or -II had no effect on thecal androstenedione production but increased ( P < 0.05) thecal progesterone production. In the presence of LH, 100 ng/ml of IGF-I increased progesterone and androstenedione production to a greater degree than did 100 ng/ml of IGF-II. Maximal effects of IGF-I and insulin on thecal steroidogenesis were similar and were not additive. Anti-IGF type I receptor antibodies attenuated ( P < 0.05) the stimulatory effect of both IGF-I and IGF-II on thecal cell steroidogenesis. Use of radioligand assays demonstrated that specific receptors for 125I-IGF-II existed in thecal cells with a 25 ng/well of IGF-II causing 50% inhibition of binding. IGF-I cross-reactivity with 125I-IGF-II receptors averaged 3% whereas cross-reactivity of IGF-II with 125I-IGF-I receptors averaged 114%. These results indicate that the stimulatory effects of IGF-II on thecal cell steroidogenesis is mediated by IGF type I receptors and thus IGF-II, like IGF-I, may play a significant role in thecal cell steroidogenesis during follicular development.