Mutant female mice carrying a singlemZP3allele produce eggs with a thin zona pellucida, but reproduce normally

Abstract

The mouse egg zona pellucida (ZP) is composed of three glycoproteins, called mZP1−3. Disruption of the mZP3 gene by targeted mutagenesis yields mice that are homozygous (mZP3^−/−) for the null mutation; although the mutant mice are viable, females are infertile and their eggs lack a ZP. On the other hand, females heterozygous (mZP3^+/−) for the mutation are fertile and their eggs have a ZP. Here, we examined fully grown oocytes from mZP3^+/− females and found that, although they have a ZP, it is less than half the width (∼ 2.7 μm; volume, ∼ 56 pl) of the ZP of oocytes from wild–type (mZP3^+/+) mice (∼ 6.2 μm; volume, ∼ 145 pl). Oocyte ZP were purified from ovarian homogenates by gradient centrifugation. Immunostaining of purified ZP on Western gels permitted an estimate to be made of the relative amounts of mZP3 and mZP2 present in the ZP of oocytes from mZP3^+/+ and mZP3^+/− mice. We found that the ZP from mZP3^+/− mice contained, on average, 55±15% of the mZP3 and 44±8% of the mZP2 present in the ZP of mZP3^+/+ mice; a result quite consistent with the observed widths and calculated volumes of the ZP. Despite the presence of a relatively thin ZP surrounding their eggs, reproduction of female mZP3^+/− mice was indistinguishable from female mZP3^+/+ mice. These results strongly suggest that, when a single mZP3 allele is present, approximately half the wild-type amount of mZP3 and approximately half the wild-type amount of mZP2 is assembled into a ZP. While this produces a relatively thin ZP, it apparently has no affect on reproduction. Furthermore, these results are consistent with the current molecular model for ZP structure.