STRUCTURE & FUNCTION OF THE EPIDIDYMIS AND VAS DEFERENS

结构

• 批准号: 6387449
• 负责人: DAVID W HAMILTON
• 金额: $ 20.27万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-29 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6387449
• 关键词: SDS polyacrylamide gel electrophoresis; acrosome; calcium channel; calcium flux; confocal scanning microscopy; cysteine; epididymis; fertilization; immunoprecipitation; laboratory mouse; laboratory rat; membrane fusion; polymerase chain reaction; protein binding; protein structure; sperm; sperm capacitation; western blottings

Funds are requested to carry out projects on the function of proteins of the Crisp superfamily in the male reproductive tract. Three Crisp genes encode proteins in an organ-specific manner: Crisp-2 is testis- specific, Crisp-3 is submandibular gland-specific and Crisp-1 is expressed primarily in the epididymis. Crisp genes are found in mammals as diverse as rats, guinea pigs, mice and humans and so must have an important function in the organs within which they are expressed. Crisp-1 proteins are synthesized and secreted by the epididymal epithelium and subsequently bind to sperm in a domain-specific manner in rats. Protein D, which binds both loosely and tightly, localizes to the head of the sperm on the plasma membrane overlying the acrosome and has been shown to play a role in fertilization. Protein E binds specifically to plasma membrane of the tail, and also participates in fertilization. Proteins D and E have significant homology to helothermine (a salivary gland toxin that blocks ryanodine receptors) and to a number of insect and snake venom proteins that are capable of regulating ion channels. We hypothesize that proteins D and E function to regulate sperm ion fluxes, particularly calcium. In the first specific aim we explore the roles of Crisp-1 during sperm-egg fusion. In the second aim, we explore their role in capacitation and the acrosome reaction, both processes that require large ion fluxes. Finally, in the last specific aim we address the question of mechanism(s) that mediate binding of protein D and E to sperm in the epididymis. We hypothesize that the signal for domain localization resides in the NH2 terminus of the molecule and the carboxyl terminus, where 14 of the 16 cysteines are found, provides the plasma membrane binding domain. These studies will contribute significant knowledge on the role of a potentially important group of proteins both in the physiology of the epididymis and in fertilization.

要求资金进行有关蛋白质功能的项目 男性生殖道的脆皮超家族。三个酥脆 基因以器官特异性编码蛋白质：crisp-2是睾丸 - 具体的，Crisp-3是下颌腺特异性和Crisp-1是 主要在附睾中表达。在哺乳动物中发现了酥脆的基因 像大鼠，豚鼠，老鼠和人类一样多样化，因此必须有一个 在表达的器官中的重要功能。 CRISP-1蛋白是由附子合成和分泌的 上皮并随后以特异性方式与精子结合 在老鼠中。蛋白质D结合松散和紧密地结合，定位于 质膜上的精子的头，上覆盖了ACROSOMOS和 已被证明在受精中起作用。 蛋白E结合 专门针对尾巴的质膜，也参与 受精。 蛋白质D和E具有重要的同源性 Helothothermine（唾液腺毒素阻断Ryanodine受体） 以及许多能够具有的昆虫和蛇毒蛋白 调节离子通道。 我们假设蛋白质D和E功能 调节精子离子通量，尤其是钙。 在第一个 具体目的，我们探讨了精子融合中Crisp-1的作用。 在第二个目标中，我们探索了它们在电容和 ACROSOM体反应，这两个过程都需要大离子通量。 最后，在最后一个具体目的中，我们解决了一个问题 介导蛋白D和E与精子与精子结合的机制 附睾。 我们假设域定位的信号 位于分子和羧基末端的NH2末端， 其中16个半胱氨酸中有14个，提供质膜 结合域。 这些研究将为有关 潜在重要的蛋白质群在 附睾和受精的生理学。