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Development of diagnosis and therapy for male infertility ; The infertility-related gene approach including seminal plasma motility inhibitor

男性不育症诊断和治疗的发展；

基本信息

批准号：
11694320
负责人：
IWAMOTO Teruaki
金额：
$ 4.67万
依托单位：
St.Marianna University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Our aim was to determine the effect of Sg on capacitation, which is the series of transformations that spermatozoa must undergo to become fertile. We induced capacitation by ultrafiltrates of fetal cord blood. It was observed that hyperactivation of the human spermatozoa was suppressed by the existence of Sg. But the dose dependence on the concentration of Sg could not be found because the analysis on the motility pattern of spermatozoa by CASA was very unstable. Using the acrosome reaction as an index, we revealed that Sg suppressed capacitation of the human spermatozoa in the dose dependent manner. By the measurement of chemiluminescence using Xantine oxidase, one mechanism of suppressed capacitation by Sg was interference with O_2 generation during this process. We also found that Sg was degraded by the incubation with the washed spermatozoa and the degradation products also inhibited capacitation of the human spermatozoa. These data suggest that Sg and/or its degradation products may be natural regulators of sperm capacitation.We investigated whether the mutation of the Sg I causes infertility. We have previously reported a deletion of 180 bp in the second exon of the Sg I in 5 infertile men and 8 fertile men, and we also found the smaller products of deleted Sg I(than normal one)in one of these fertile men. The mutation form protein is currently being purified and analyzed to determine the difference in the protein function. Though the SNP analysis of the Sg gene is carried out by the DNA two-dimensional electrophoretic method, presently, it has not been confirmed. It succeeded in the expression of the r-Sg protein with much high activity and high-purity. The antibody that produced this r-Sg as an antigen peculiarly reacted with native Sg and SPMI.

我们的目的是确定Sg对获能的影响，获能是精子必须经历的一系列转化才能生育。我们用胎儿脐带血的超滤液诱导获能。观察到Sg的存在抑制了人精子的过度激活。但由于CASA对精子运动模式的分析很不稳定，未发现Sg浓度与剂量的依赖关系。以顶体反应为指标，我们发现Sg以剂量依赖的方式抑制人精子的获能。用黄嘌呤氧化物酶测定其化学发光表明，Sg抑制获能的机制之一是干扰了这一过程中O2的产生。我们还发现，Sg与洗涤的精子孵育后被降解，降解产物也抑制了人类精子的获能。这些数据表明，Sg和/或其降解产物可能是精子获能的天然调节因素。我们调查了Sg I突变是否导致不育。我们已经报道了5名不育男性和8名生育男性Sg I基因第二外显子180个碱基的缺失，我们还在其中一名生育男性中发现了Sg I缺失的产物(比正常产物更小)。突变形式的蛋白质目前正在进行纯化和分析，以确定蛋白质功能的差异。虽然Sg基因的SNP分析是通过DNA双向电泳法进行的，但目前还没有得到证实。成功地表达了高活性、高纯度的r-Sg蛋白。产生这种r-Sg作为抗原的抗体与天然Sg和SPMI发生特异性反应。