ABSTRACT. Two pony mares were immunized against recombinant porcine inhibin α subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 ± 4.30, 2.75 ± 0.69 and 2.51 ± 0.63 vs 6.50 ± 1.43, 1.83 ± 0.44 and 1.33 ± 0.38, respectively), but the ovulation rate remained unchanged after immunization. The average plasma concentration of FSH and estradiol-17β during the estrous cycle increased significantly (P<0.05) after immunization. These results suggest that immunization against inhibin is a useful tool to increase the number of ovarian follicles during the estrous cycle of pony mares. Moreover, the present study supported the concept that inhibin plays a major role in the control of follicular growth through its inhibitory effect on FSH secretion synergistically with steroid hormones. KEY WORDS: follicular development, FSH, immunization, inhibin, mare.J. Vet. Med. Sci. 66(1): [31][32][33][34][35] 2004 The physiological mechanisms responsible for limiting one follicular development and ovulation from multiple follicles during the normal follicular phase in the mare is not yet fully understood. Attempts at inducing multiple ovulation or promoting fertility in the mare with various treatments such as equine chorionic gonadotropin (eCG), equine pituitary extract and PGF 2 α have met with limited success [2,6,10,12,17,20]. Currently, crude pituitary extracts are the most reliable method for inducing multiple ovulations in the mare and this method usually results in one to four ovulations per cycle [5], but pituitary extracts are not readily available and the regimens required are relatively intensive.Inhibin is a large glycosylated protein consisting of two subunits linked by disulfide bridges, and specifically inhibits FSH secretion and suppresses the expression of GnRH receptors in the pituitary gland [1,7,15,22]. On the other hand, the production of inhibin is stimulated by FSH thereby forming a tight feedback circuit [18,19]. It is argued that ovarian inhibin may be the most important hormone for determining the species-specific number of ovulations in both single and multiple ovulatory species.In cattle, active immunization against inhibin enhanced ovarian follicular development and the number of oocytes collected by transvaginal follicular aspiration [8,9]. Furthermore, passive immunization of mares against inhibin during the follicular phase of the estrous cycle results in a marked increase in plasma concentrations of FSH induced by immunoneutralization of endogenous inhibin, and stimulates the rapid growth of a large number of follicles. As a result, multiple ovulations can be induced in mares [16].This study...