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Heteroimmunization with isolated pig zonae pellucidae

Treatment of zonae pellucidae with antiserum containing antibody activity against zona-specific antigens produces a precipitation layer on the outer surface of the zona (Ownby & Shives, 1972; Sacco & Shivers, 1973; Sacco, 1977) which inhibits both sperm attachment and penetration (see Shivers, 1974, for review). The presence of this precipitation layer is believed to be responsible for the observed inhibition of fertilization in vitro and in vivo following either antiserum treatment of eggs or passive immunization (Jilek & Pavlok, 1975; Oikawa & Yanagimachi, 1975; Tsunoda & Chang, 1976a, b, c). To date, all antisera demonstrating anti-zona activity have been prepared by immunizing animals with ovary, unfertilized eggs or preimplantation-stage embryos (see Shivers, 1974, 1975 for review). These antisera initially contain antibodies directed against many tissue components and must be rendered specific to the zona by absorption procedures. Such absorbed antisera are obviously contaminated by components of the absorbing tissues which may interfere with or influence results from subsequent antifertility testing. We report here initial attempts to produce monospecific anti-zona pellucida serum by using isolated pig zonae obtained from unfertilized follicular eggs as the immunizing antigen. By the use of isolated zona material as the immunogen rather than ovary or intact eggs or embryos, the resulting antiserum may not require absorption to be monospecific for zona pellucida.