Analysis Tools
reproductive system
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• ovarian histology of mice expressing human ZP2 in place of mouse ZP2 appears normal
• zonae pellucidae of oocytes from mice expressing human in place of mouse ZP2 appear similar histologically to those of wild-type mice; the human ZP2 protein and mouse ZP1 and ZP3 proteins are immunohistologically detectable throughout the zona matrix
• cumulus-free ovulated oocytes expressing human instead of mouse ZP2 bind mouse sperm, but fail to bind human sperm
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• female mice expressing human in place of mouse ZP2 ovulate fewer oocytes than do wild-type or ZP2/ZP3 rescue females when primed with pregnant mare serum and stimulated with human chorionic gonadotrophin
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• although expression of human ZP2 restores fertility to otherwise sterile female mice lacking mouse ZP2, their average litter size is smaller than that of wild-type controls
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embryo
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• one- and two-celled embryos having human in place of mouse ZP2 exhibit persistent sperm binding, even after multiple washes using a wide-bore pipet
• cleavage of mouse ZP2 following fertilization is demonstrated by increased SDS-PAGE mobility of the reduced protein in the wild-type two-cell embryo versus in the ovulated oocyte; this mobility shift fails to occur in two-cell embryos in which human ZP2 replaces its mouse ortholog
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• 24 hours after in vitro insemination, only 68% of ovulated oocytes in cumulus mass from female mice expressing human in place of mouse ZP2 had progressed to the 2-cell stage, versus 82% of wild-type eggs
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