SPERM FUNCTION IN FERTILIZATION EVENTS

精子的质膜和顶体膜被划分成不同的区域。 具有执行特定功能的不同的分子组成 哺乳动物受精所必需的。这样做的长期目标是 建议是定义组装离散的分子过程 膜结构域及其生化调控机制的研究 顶体周围质膜与外膜的融合 顶体反应过程中的顶体膜。提出了四个目标 解决这些目标。第一个目的是确定该系统的功能 精子顶体周围质膜蛋白PM52(精子) 获能和/或顶体反应。免疫沉淀和免疫印迹 将执行重叠分析以确定 PM52的细胞质结构域和Pro-2的模拟多肽 将测试PM52的富含结构域对获能的影响，以及 顶体反应。目标2是确定对接相互作用是否 顶体周围质膜与顶体外部之间 膜产生结构域特异的蛋白质定位。免疫印迹 而印迹覆盖层将识别外部的PM52结合蛋白 顶体细胞膜和免疫荧光实验将测试 PM52-顶体外膜相互作用的中断导致 蛋白质重新分配。目标3是确定生殖细胞亚型是否 突触素和突触素在神经细胞膜融合事件中的作用 顶体反应。免疫定位和免疫沉淀分析 以确定这些蛋白质是否组装成一个 顶体胞吐所需的膜融合复合体。目标4是 确定蛋白质-蛋白质之间的相互作用，调节 顶体外膜-顶体基质复合体。生化和 将进行超微结构实验以鉴定 顶体外膜，结合和调节顶体的外膜 顶体基质组件在顶体内的分布。 这些研究的完成将为大会提供新的见解 哺乳动物顶体节段及其功能的研究 受精和研究结果具有潜在的战略应用价值 用于调节和/或改善生育能力。

The sperm plasma and acrosomal membranes are partitioned into domains of distinct molecular composition which perform specific functions required for mammalian fertilization. The long range goals of this proposal are to define the molecular processes which assemble discrete membrane domains and to identify the biochemical mechanisms regulating the fusion between the periacrosomal plasma membrane and the outer acrosomal membrane during the acrosome reaction. Four aims are proposed addressing these goals. Aim one is to identify the function of the periacrosomal plasma membrane protein PM52 (sperad) in sperm capacitation and/or the acrosome reaction. Immunoprecipitation and blot overlay analyses will be performed to identify ligands for the cytoplasmic domain of PM52 and permeant peptide mimetics of the proline- rich domain of PM52 will be tested for effects on capacitation and the acrosome reaction. Aim 2 is to determine if docking interactions between the periacrosomal plasma membrane and the outer acrosomal membrane generate domain-specific protein localization. Immunoblotting and blot overlays will identify PM52 binding proteins in the outer acrosomal membrane and immunofluorescence experiments will test if disruption of PM52-outer acrosomal membrane interaction results in protein redistribution. Aim 3 is to determine if germ cell isoforms of syntaxin and synaptophysin function in the membrane fusion events of the acrosome reaction. Immunolocalization and immunoprecipitation analyses will be performed to determine if these proteins assemble into a membrane fusion complex required for acrosomal exocytosis. Aim 4 is to identify protein-protein interactions regulating the assembly of the outer acrosomal membrane-acrosomal matrix complex. Biochemical and ultrastructural experiments will be performed to identify proteins of the outer acrosomal membrane which bind to, and regulate the distribution of, acrosomal matrix assemblies within the acrosome. Completion of these studies will provide new insights into the assembly of the acrosomal segment and of its functions during mammalian fertilization and the findings have potential application to strategies for fertility regulation and/or improvement.