Immunization against an inhibin subunit produced by recombinant DNA techniques results in increased ovulation rate in...

Seven Merino-Border Leicester cross-bred ewes were immunized with a purified fusion protein, produced by recombinant DNA methods, of the alpha subunit of bovine inhibin. Four animals were immunized with the fusion protein alone and three with a conjugate made by coupling the fusion protein to keyhole limpet haemocyanin (KLH) using glutaraldehyde. Each animal received four injections of the fusion protein over 93 days. The animals were synchronized using progestagen sponges and subjected to laparoscopy for the determination of ovulation rates in two consecutive cycles (days 115 and 135). The immunized animals had overall mean ovulation rates for each cycle of 3.4 and 3.4 which was significantly (P less than 0.001) above the rates of 1.1 and 1.4 determined for the controls, which had either received no treatment (n = 5) or had been immunized with 300 micrograms KLH (n = 4). Analysis of antisera taken on day 115 showed significant fusion protein antibodies and iodinated inhibin-binding capacity in the test but not control groups. Furthermore, antisera to the fusion protein in four out of seven ewes neutralized the inhibin bioactivity of ovine follicular fluid in an in-vitro bioassay. These data demonstrate that neutralization of inhibin can be effected by immunization with bovine inhibin alpha subunit and that such immunization results in increased ovulation rates as predicted from the biological role of inhibin as a suppressor of FSH.