SPERM FUNCTION IN FERTILIZATION EVENTS

受精事件中的精子功能

• 批准号: 3318477
• 负责人: GARY E OLSON
• 金额: $ 9.76万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3318477
• 关键词: acrosome; adenosinetriphosphatase; antibody; cell membrane; cytoskeleton; electron microscopy; fertilization; gel electrophoresis; immunochemistry; intracellular membranes; membrane activity; sperm

The long-range objectives of this proposal are to identify the structural and biochemical properties of specific membrane domains and membrane-associated assemblies of mammalian spermatozoa and to determine how these membrane domains and membrane-associated assemblies function during the fertilization process. In the present proposal attention will be focused upon the membranes participating in the acrosome reaction, in order to identify mechanisms contributing to the membrane fusion process, and upon the plasma membrane of the postacrosomal segment, in order to characterize its role in gamete fusion. The specific objectives of this proposal are 1) to characterize the macromolecular complex (fuzzy-coat) which is exclusively localized on the luminal face of the outer acrosomal membrane; 2) to determine how the fuzzy-coat complex associates with integral membrane proteins of the outer acrosomal membrane; 3) to define the role of the fuzzy-coat complex in the membrane vesiculation process of the acrosome reaction; 4) to utilize antibodies against the fuzzy-coat complex to identify acrosome reacting sperm; 5) to determine if the fuzzy-coat assembly is modified during sperm maturation in the epididymis; 6) to identify enzyme activities associated with membrane domains participating in the acrosome reaction; 7) to characterize the postacrosomal sheath; and 8) to identify the intracellular distribution of cytoskeletal proteins in mammalian spermatozoa. Nitrogen cavitation and selective extraction procedures will be employed to disrupt spermatozoa and purified subcellular fractions consisting of specific membrane domains or membrane-associated assemblies will be isolated by a variety of centrifugation protocols. A combination of ultrastructural, biochemical and immunological techniques, which have previously been utilized to identify the functional properties of membrane-associated assemblies in other cell types, will be employed to pursue these specific aims. A resolution of these specific aims will contribute to our understanding of membrane physiology, to our understanding of major events in the fertilization process and to male reproductive biology.

本提案的长期目标是确定结构性 和特定膜结构域的生物化学性质， 哺乳动物精子的膜相关组件，并确定 这些膜结构域和膜相关组件是如何发挥作用的 在受精过程中。 在本建议中，将注意 关注参与顶体反应的膜， 为了鉴定有助于膜融合过程的机制， 和顶体后节的质膜上，以便 描述其在配子融合中的作用。 本提案的具体目标是：（1） 大分子复合物（绒毛涂层），仅位于 顶体外膜的腔面; 2）确定 绒毛膜复合物与外膜的完整膜蛋白结合 顶体膜; 3）确定绒毛膜复合体在顶体膜中的作用。 顶体反应的膜泡化过程; 4）利用 抗绒毛膜复合物抗体，以识别顶体反应 精子; 5）确定在精子过程中绒毛被膜组件是否被修改 附睾成熟; 6）鉴定与附睾成熟相关的酶活性 膜结构域参与顶体反应; 7） 表征顶体后鞘;和8）鉴定细胞内 哺乳动物精子中细胞骨架蛋白的分布。 氮 将采用空化和选择性提取程序来破坏 精子和纯化的亚细胞级分， 膜结构域或膜相关组件将通过 各种离心方案。 结合超微结构， 生物化学和免疫学技术，这些技术以前被 用于识别膜相关的功能特性， 其他细胞类型的组装，将被用来追求这些特定的 目标。 解决这些具体目标将有助于我们 对膜生理学的理解，对我们理解重大事件 受精过程和雄性生殖生物学。