To clarify the mechanism of FSH secretion from the pituitary induced by activin A, we studied the interaction between activin A and gonadal steroids in inducing FSH release from primary cultured female rat pituitary cells in serum-free medium. The basal release of FSH was stimulated by activin A, testosterone (T) and progesterone (P), and T and P also facilitated basal FSH release stimulated by activin A. The GnRH-stimulated FSH release was facilitated by activin A, P and 17 beta-estradiol (E2), but suppressed by T. The effect of activin A on GnRH-stimulated FSH release was facilitated by P, but not affected by T or E2. These findings suggested that the interaction between activin A and T or P may be involved in the regulation of FSH secretion during the estrus cycle in rats.