Suckling causes a delay in onset of estrus and ovulation in cattle postpartum. In addition, the suckling stimulus causes the release of corticoids presumably via ACTH. Since any hormone released by suckling is a potential inhibitor of gonadotropin secretion and/or ovulation, we investigated the effects of ACTH and cortisol on LHRH-induced LH release from bovine pituitary cells in vitro. Anterior pituitary glands were obtained from cows killed during the luteal phase of an estrus cycle (days 5-15). Pituitary cells, disaggregated enzymatically, were grown in Dulbecco's medium containing 10% dextran charcoal-stripped fetal calf serum. On day 5, cultures were washed and reincubated in serum free medium containing the hormone being tested. After 6 h of incubation, LHRH was added in 10 microliters medium and incubation continued for an additional 6 h. ACTH at 4.3 X 10(-9), 4.3 X 10(-8), and 4.3 X 10(-7) M had no effect on basal or LHRH-induced LH release. Cortisol at 12.1 ng/ml decreased (P less than 0.001) the slope of LHRH response curve (b1 = 2.9 vs. 5.5 for controls). To determine if this effect was specific for cortisol, we compared cortisol, dexamethasone, and progesterone. LHRH-induced LH release (percent of control) was decreased (P less than 0.001) by 12.1 ng/ml cortisol (98%), 1, 5, and 10 ng/ml dexamethasone (60%, 71%, and 88%), but not by 3.1 ng/ml progesterone. The inhibitory effect of cortisol was reversible. Thus, LHRH-induced LH release (percent of controls) at 0, 24, 48, and 72 h after a 24-h exposure to 12.1 ng/ml cortisol was 19%, 89%, 100%, and 115%, respectively. We have demonstrated that cortisol at concentrations found normally in blood of cows postpartum will inhibit LHRH-induced LH release from bovine pituitary cells. This observation is consistent with the hypothesis that cortisol released by suckling may inhibit gonadotropin secretion postpartum and as such may prolong the anovulatory interval postpartum.