An ovarian perifusion technique was used to determine if there is a direct suppressive effect of PRL on human gonadal steroid secretion. Ovarian tissue from nine patients was examined. Ovine PRL (0.1-10 IU/ml) directly suppressed progesterone and 17 beta-estradiol secretion by human ovaries. hCG (1-10 IU/ml) stimulated both progesterone and 17 beta-estradiol secretion. Simultaneous administration of PRL (5-10 IU/ml) suppressed the stimulatory effect of hCG. The ovarian tissue obtained from a hyperprolactinemic patient did not respond to hCG. These results indicate that PRL inhibits both basal and gonadotropin-stimulated ovarian steroid secretion by human ovaries and that this may be one cause of the hypogonadism associated with hyperprolactinemia.