Inhibition of in vivo fertilization by active immunization of male hamsters against a 26-kDa sperm glycoprotein

Abstract

We have identified a 26-kDa (P26h) epididymal hamster sperm glycoprotein with a species-specific affinity for zona pellucida glycoprotein. Two immunological procedures have been used to document the biological function of this sperm component; active immunization of males against P26h and inhibition of sperm-zona pellucida binding in vitro by anti-P26h antibodies. The immunized male hamsters produced circulating antibodies specific to P26h. Indirect immunofluorescence studies showed that these antibodies bind to the surface of the sperm covering the acrosome. These males were mated with superovulated females, and although spermatozoa were recovered from the genital tract, none of the 194 oocytes recovered were fertilized. In contrast, control males immunized with hamster albumin fertilized 97.4% of the oocytes. Unlike control spermatozoa, those recovered from the cauda epididymidis of males immunized with P26h were characterized by the presence of antibodies at the surface of the acrosome. To establish whether the inhibition of in vivo fertilization by active immunization was occurring at the level of sperm-zona pellucida interaction, a polyclonal antiserum against P26h was raised, and the IgG fraction was added to an in vitro sperm-zona pellucida assay. Compared to the preimmune serum, the IgG inhibited the binding of spermatozoa in a dose-dependent manner. The Fab fragments generated from these IgGs were almost as efficient in inhibiting the binding. These results are discussed with regard to a possible function of P26h in hamster gamete interaction.