Glycan-Independent Gamete Recognition Triggers Egg Zinc Sparks and ZP2 Cleavage to Prevent Polyspermy.

The zona pellucida surrounding ovulated eggs regulates monospermic fertilization necessary for successful development. Using mouse transgenesis, we document that the N terminus of ZP2 is sufficient for sperm binding to the zona matrix and for in vivo fertility. Sperm binding is independent of ZP2 glycans and does not occur after complete cleavage of ZP2 by ovastacin, a zinc metalloendopeptidase stored in egg cortical granules. Immediately following fertilization, a rapid block to sperm penetration of the zona pellucida is established that precedes ZP2 cleavage but requires ovastacin enzymatic activity. This block to penetration is associated with release of zinc from cortical granules coincident with exocytosis. High levels of zinc affect forward motility of sperm to prevent their passage through the zona matrix. This transient, post-fertilization block to sperm penetration provides a temporal window to complete the cleavage of ZP2, which prevents sperm binding to ensure monospermy.

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