Current evidence favors the hypothesis that granulosa cell-derived basic fibroblast growth factor (bFGF) may be the centerpiece of an intraovarian autocrine loop. In this report we examine the possibility that bFGF may also be involved in paracrine interactions at the level of the ovarian theca-interstitial cell. To this end, whole ovarian dispersates obtained from immature rats were cultured for 96 h under serum-free conditions. The accumulation of androsterone, the major androgenic steroid detected, increased 3- to 5-fold over baseline in response to treatment with hCG (1 ng/ml), an effect further optimized (2- to 4-fold) by supplementation with insulin-like growth factor-I (10 ng/ml), insulin (1 microgram/ml), terbutaline (10(-6) M), or high density lipoprotein (100 micrograms/ml). In the absence of these optimizing supplements, bFGF was without effect on basal androsterone accumulation, but produced a relatively modest (20%) inhibition of hCG hormonal action. In contrast, bFGF proved a highly potent inhibitor (80%) of hCG-stimulated androgen biosynthesis in supplement-enriched cultures. This reversible bFGF action proved to be time and dose dependent, with a minimal time requirement of 48 h and a median inhibitory dose of 2 ng/ml. Unaccounted for by altered (hCG-stimulated) cAMP generation or a diminution in the viable cell mass, the antigonadotropic effect of bFGF may by inference be assumed to involve a site(s) of action distal to or independent of cAMP generation. In this connection, cellular radiolabeling with [3H] pregnanolone (3 alpha-hydroxy-5 alpha-pregnane-20-one) revealed bFGF to be a potent inhibitor of the steroidogenic enzyme 17 alpha-hydroxylase/17-20-lyase. As such, these findings are in keeping with the possibility that locally derived bFGF may exert a dual inhibitory action on (mature) ovarian estrogen production by reducing androgen substrate provision as well as by exercising its now established ability to attenuate granulosa cell aromatase activity.