Zona pellucida glycoprotein mZP3 produced in milk of transgenic mice is active as a sperm receptor, but can be lethal to newborns.

Mouse egg zona pellucida glycoprotein mZP3 (approximately 83 kDa M(r)) serves as a species-specific sperm receptor and acrosome reaction-inducer during fertilization in mice. These biological activities are dependent on certain mZP3 serine/threonine- (O-) linked oligosaccharides present at the combining-site for sperm. In an attempt to produce large amounts of biologically active mZP3, we generated several transgenic mouse lines carrying the full-length mZP3 gene fused to the beta-casein gene promoter and transcription termination sequence. We found that different transgenic mouse lines have different amounts of recombinant mZP3 (approximately 63 kDa M(r)) in milk of lactating females, from approximately 0.3 to 3.5 micrograms/microliter of milk. In all cases, purified milk-mZP3 is active as a sperm receptor and acrosome reaction-inducer in vitro. Unexpectedly, we also found that development of litters from these transgenic mice is related to the amount of mZP3 in the mother's milk. In the most extreme case, litters from the highest expressers fail to live beyond about day-7 post partum unless placed immediately after birth with surrogate wild-type mothers. Litters from lower expressers initially display a complex phenotype that includes effects on hair and body growth, but some of the mice survive and, in time, are restored to a wild-type phenotype. These results demonstrate that relatively large amounts of biologically active mZP3 can be produced in transgenic mouse milk for structural and other studies, but that the presence of mZP3 in milk has dramatic developmental effects on litters, ranging from retarded hair and body growth to death of newborn pups.