This study was carried out to examine the involvement of inhibin in the regulation of FSH secretion during the growth phase of the dominant follicle in the early luteal phase of cows. Six cows were given a single i.v. bolus injection of 100 ml inhibin antiserum raised against bovine 32-kDa inhibin in a castrated male goat, and five animals received the same amount of castrated male goat serum (control serum) on Day 5 of the estrous cycle (Day 0 = estrus). All animals in each group experienced a wave of follicular development after ovulation, and the dominant follicle was over 8.5 mm in diameter on Day 5. The corpus luteum was identified for each group on Day 5. Plasma concentrations of estradiol and progesterone gradually increased after ovulation and reached around 3.7 pg/ml and 3.0 ng/ml on Day 5, respectively, suggesting that the dominant follicle and corpus luteum were functional. Administration of inhibin antiserum produced a clear increase (P < 0.001) in plasma FSH within 8 hr compared with that in control animals. Plasma levels of luteizing hormone showed a moderate increase during 40 hr after the injection of antiserum (P = 0.08). A large number of antral follicles (4 mm in diameter) developed after the hypersecretion of FSH, coupled with the rise in plasma estradiol levels. These results clearly demonstrated that inhibin neutralization during the early luteal phase produces hypersecretion of FSH with a coincident stimulation of follicular development, indicating that inhibin is an important factor for the negative regulation of FSH secretion during the early luteal phase when secretion of estradiol and progesterone are normally high.