A Study of abnormal sperm-egg interaction in male infertility

我们一直在利用人CD46从临床角度或从基础角度利用小鼠CD46研究精子-卵子相互作用异常所致的不育症，并在1997年和1998年按计划取得了完美的结果。人精子CD46CD46的分子克隆具有多种器官特异性变异体，其中精子CD46无糖链，定位于顶体内膜。我们克隆了精子CD46，并展示了其特有的胞浆结构域。在CHO细胞中的瞬时表达证实了CD46在内质网和金黄地鼠实验中的定位表达，表达的精子CD46表现出比其他CD46更强的精卵相互作用。胞浆结构域在精子CD46.2的翻译后修饰中起着重要作用。小鼠CD46的分子克隆我们以人、猪和豚鼠的同源序列为模板，从小鼠睾丸cDNA文库中克隆了1.2kb的全长cDNADNA。Northern blotting和Western blotting检测到小鼠CD46在睾丸中有特异性表达。我们正在用表达的小鼠CD46进行功能分析，敲除CD46的小鼠将阐明其在精子与卵子相互作用中的作用。

We have been studying infertility due to abnormal sperm egg interaction either from clinical aspect using human CD46 or from basic aspect using murine CD46 and in 1997 and 1998, we obtained perfect results as we scheduled.1. Molecular cloning of human sperm CD46CD46 has many organ-specific variants and sperm CD46 is very characteristic, which has no sugar chain and localized in inner-acrosomal membrane. We cloned sperm CD46 and showed its characteristic cytoplasmic domain. Transient expression in CHO cells proved localized expression of CD46 without sugar chain in endoplasmic reticulumn and in Hamster test, the expressed sperm CD46 revealed stronger sperm-egg interaction than the other CD46. Cytoplasmic domain has an important role in post-translational modification in sperm CD46.2. Molecular cloning of murine CD46We cloned full length 1.2kb cDNA from murine testis cDNA library using PCR product as a probe, which primers are from homologous sequences among human, pig and guinea pig. Northern blotting and Western blotting using antibody to synthesized peptide revealed testis specific expression of murine CD46. We are doing functional analysis using expressed murine CD46 and knock out mouse of CD46 will clarify its role in sperm egg interaction.