Characterization of Human Sperm Binding to Homologous Recombinant Zona Pellucida Proteins

The union between mammalian gametes begins with the sperm binding to the zona pellucida (ZP). We studied the interaction between human sperm and ZP by using recombinant human ZP proteins (rhZP). The cDNAs coding for human ZP2, ZP3, and ZP4 were expressed in Sf9 cells and proteins were characterized to determine their competence for sperm binding. Capacitated human sperm binding abilities were analyzed using immobilized rhZP and a well-characterized antihuman sperm antiserum. The results demonstrated that all rhZP proteins were structurally similar to their native counterparts and were specifically recognized, in a dose and time dependent manner, by human sperm. The rhZP4 was the main sperm binder followed by rhZP3 and rhZP2, although combinations of rhZP proteins enhanced sperm adhesion. Moreover, this experimental approach may represent a useful model to study sperm-ZP interactions for research and clinical purposes.