The activity of 17alpha-hydroxylase/C17-C20 lyase (17alpha-hydroxylase) in the ovaries and steroid hormone levels in the plasma were studied in immature hypophysectomized rats (IH-rats) treated with human recombinant follicle-stimulating hormone (rec-FSH) alone or combined with various doses of human chorionic gonadotropin (hCG). Eleven days after hypophysectomy, rats were given rec-FSH alone (total dose, 40 IU), or combined with various doses (0.1 to 10 IU in total) of hCG, twice daily for 4 days. Plasma levels of progesterone, testosterone and estradiol, and ovarian 17alpha-hydroxylase activity were measured 18 h after the last injection. Histology showed that the ovaries treated with rec-FSH alone had large antral follicles, the theca interna cells of which were small in size compared with those treated with rec-FSH combined with hCG. The activity of 17alpha-hydroxylase in the ovaries of IH-rats treated with rec-FSH alone was lower than that in the control IH-rats. It was markedly increased by treatment with rec-FSH combined with 1 or 10 IU hCG. Immunohistochemistry revealed that 17alpha-hydroxylase was localized only in the oocyte in the ovaries of control IH-rats and those treated with rec-FSH alone. When the IH-rats were treated with rec-FSH plus hCG, the number of immunopositive theca interna cells and interstitial cells, and their immunointensity were increased in an hCG dose-dependent manner. The plasma estradiol levels in the IH-rats treated with rec-FSH alone were low, but significantly higher than those in the control IH-rats, and estradiol levels were noticeably elevated in IH-rats treated with rec-FSH plus hCG. These results suggest that a synergism between hCG (LH) and FSH is essential for follicular development and steroidogenesis in the ovaries, implying paracrine effects among granulosa cells, theca cells and probably oocytes. The functional significance of 17alpha-hydroxylase in the oocytes is discussed in relation to estradiol production.