Studies on Sperm Function in Fertilization

为了在哺乳动物fertilliation中实现精子功能（精子/蛋交互），我们使用基因改良的突变老鼠进行了生物化学和生物实验，并获得了后续结果：1）利用了前列腺素在fertilization中的作用，我们已经审查了前列腺素在前列腺素反应中的作用，并使用了前列腺素-非修饰的突变老鼠。当精液到达（附着）时，并与无冠卵的区域内的无冠卵被评估为体外，而在野生类型和交叉不合格的老鼠之间没有明显的区别。免疫化学分析演示了几种交叉蛋白质从Acrye D2-/-D2小鼠sperm在钙离子体和溶解区Pellucida诱导的交叉体反应中的释放，在正常膜血管中观察到的交叉体反应。这些数据表明，在acrosin-deficient中，区域皮卢西达的延迟渗透率 ... More 小鼠根据蛋白质从染色体组中分离的变化率得出的结果，并提供了第一个证据，即Acrosin的主要作用可以加速acrosome反应期间分离的acrosomal组分（s），以识别一种新颖的候选人（s），其中一个DNA片段是从acrosin-deficient小鼠测试的cDNA库中扩增出来的。然后使用一个探针来显示小鼠测试的cDNA库。Complementary DNA克隆编码每个四个类似但不同的序列蛋白质，TESP 1，TESP 2，TESP 3， and TESP 4， have been identified. 3） We produced transgenic mouse lines that accumulate mutated绿色荧光蛋白质（EGFP） in sperm acrosome。需要时间来说明分散的交叉成分在大约3秒后在交叉组反应的同一时间。4）人类表皮状瘤的主要秘密蛋白质的脊髓同源性，HE 1从cauda表皮状流体中被净化。HE 1同源物被发现是一种主要的胆固醇结合蛋白，并建议它被纳入了表皮基质中的脂质组成的规则中。Less（低）

To elucidate the sperm function (sperm/egg interaction) in mammalian fertilization, we have carried out biochemical and biological experiments using genetically modified mutant mice, and have obtained the following results :1) To elucidate the role of acrosin in fertilization, we have examined the involvement of acrosin in the acrosome reaction of sperm, using the acrosin-deficient mutant mice. When the ability of sperm to adhere (attach) and bind to the zona pellicida of cumulus-free eggs was assessed in vitro, no significant difference was observed between the wild-type and acrosin-deficient mice. Immunocytochemical analysis demonstrated that the release of several acrosomal proteins from the acrosome of AcrィイD2-/-ィエD2 mouse sperm was significantly delayed during the calcium ionophore- and solubilized zona pellucida-induced acrosome reaction, in spite of normal membrane vesiculation. These data indicate that the delayed sperm penetration of the zona pellucida in the acrosin-deficient … More mouse results from the altered rate of protein dispersal from the acrosome, and provide the first evidence that the major role of acrosin is to accelerate the dispersal of acrosomal components during acrosome reaction.2) To identify a novel candidate(s) for acrosomal proteins that act on the sperm/egg interaction, a DNA fragment was PCR-amplified from a cDNA library of acrosin-deficient mouse testis, and then used as a probe to screen a mouse testis cDNA library. Complementary DNA clones encoding each of four similar but different serine proteases, TESP1, TESP2, TESP3, and TESP4, have been identified.3) We produced transgenic mouse lines that accumulate mutated green fluorescent protein (EGFP) in sperm acrosome. The time required for the dispersal of acrosomal components was demonstrated to be approximately 3 seconds after the onset of acrosome reaction.4) A porcine homologue of the major secretory protein of human epididymis, HE1 was purified from cauda epididymal fluids. The HE1 homologue was found to be a major cholesterol-binding protein, suggesting that it is involved in the regulation of the lipid composition of the sperm membranes during sperm maturation in epididymis. Less