A Study of abnormal sperm-egg interaction in male infertility

我们一直在研究精子-卵子相互作用异常引起的不育症，无论是从临床方面使用人CD 46，还是从基础方面使用小鼠CD 46，并在1997年和1998年，我们获得了理想的结果，正如我们计划的那样。人精子CD 46分子克隆CD 46具有多种器官特异性变异，其中精子CD 46具有很强的特征性，不含糖链，定位于顶体内膜。我们克隆了精子CD 46，并显示了其特征性的胞质结构域。在CHO细胞中的瞬时表达证实了不含糖链的CD 46在内质网中的定位表达，并且在体外试验中，表达的精子CD 46显示出比其它CD 46更强的精卵相互作用。胞浆区在精子CD46.2的翻译后修饰中起重要作用。小鼠CD_（46）的分子克隆以人、猪和豚鼠的同源序列为引物，以PCR产物为探针，从小鼠睾丸cDNA文库中克隆了全长1.2kb的cDNA。北方印迹和Western印迹显示小鼠睾丸特异性表达CD 46。我们正在利用表达的小鼠CD 46进行功能分析，并将CD 46基因敲除小鼠，以阐明其在精卵相互作用中的作用。

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.