SPERM ION CHANNEL REGULATION

精子离子通道调节

• 批准号: 2205161
• 负责人: Harvey M Florman
• 金额: $ 19.43万
• 依托单位: WORCESTER FOUNDATION FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2205161
• 关键词: acrosome; biological signal transduction; calcium channel; cell membrane; digital imaging; egg /ovum; electrophysiology; exocytosis; fertilization; fluorescence microscopy; fluorescence spectrometry; gene expression; glycoproteins; laboratory mouse; membrane permeability; membrane potentials; single cell analysis; sperm; spermatogenesis; voltage /patch clamp

ZP3, a glycoprotein constituent of the, egg's zona pellucida, regulates the initiation of sperm acrosomal exocytosis during mammalian fertilization. The long range goal of this laboratory is to understand the mechanism of ZP3 signal transduction. The current phase of this research focuses on the regulation of sperm ion channels by ZP3 as well as on the relationship between ion channel activation and the elevations of internal Ca,+ that are required for exocytosis. These studies utilize digital image processing-enhanced fluorescence microscopy, fluorometry, and patch electrode-voltage clamping in order to explore ion channel regulation in living sperm. There are four specific aims. 1) The ionic basis of sperm membrane potential will be established. These studies will involve selective ion depletion and antagonist treatments to determine the principal ion permeabilities that establish membrane potential in resting cells. 2) The effects of ZP3 on sperm membrane potential will be determined, by identifying ion channels that are activated by ZP3. 3) The role of ZP3-dependent ion channels in the induction of the acrosome reaction will be defined. Selective antagonists will be used to determine the relationship between ZP3-dependent ion channels and internal Ca,+ elevations. 4) The expression during spermatogenesis of ion channels that determine resting membrane potential or that are activated by ZP3 will determined. Health problems are posed when fertilization is either unbounded or is impeded. In this regard, the acrosome reaction is an important regulatory event during mammalian gamete interaction, with unregulated exocytosis associated a failure of fertilization. These studies define sperm ion channels that are essential components of the ZP3 signaling pathway and may provide new targets for the control of fertilization. Highly-resolved profiles of these channels are provided by state-of-the-art fluorescent and electrophysiological methods and may permit rationale design of new antagonists. In addition, identification of spermatogenic stages that express ZP3-regulated channels sets the stage for the molecular cloning and subsequent analysis of channel cDNAs.

ZP3是鸡蛋的Zona pellucida的糖蛋白成分 哺乳动物期间精子诊断性胞吐作用的启动 受精。 该实验室的远程目标是了解 ZP3信号转导的机制。当前阶段 研究侧重于ZP3的精子离子通道调节 关于离子通道激活与高程之间的关系 胞吞作用所需的内部Ca+。 这些研究利用 数字图像加工增强荧光显微镜，荧光测定法， 和贴片电极电压夹紧以探索离子通道 活精子中的调节。 有四个具体目标。 1）将建立精子膜电位的离子基础。这些 研究将涉及选择性离子耗竭和拮抗剂治疗 确定建立膜的主要离子渗透率 静止细胞的潜力。 2）ZP3对精子膜电位的影响将通过 识别ZP3激活的离子通道。 3）ZP3依赖性离子通道在诱导质体中的作用 反应将定义。选择性拮抗剂将用于确定 ZP3依赖性离子通道与内部CA+之间的关系 海拔。 4）确定离子通道的精子发生过程中的表达 将确定静息膜电位或被ZP3激活的电位。 当受精是无限或是 阻碍。 在这方面，首席反应是重要的调节 哺乳动物配子相互作用期间的事件，胞吐病不受管制 相关的受精失败。 这些研究定义了精子离子 是ZP3信号通路的必需组件的通道和 可以为控制受精的新目标提供新的目标。高度分辨 这些通道的概况由最先进的荧光灯提供 和电生理方法，并且可以允许新的原理设计 对手。此外，鉴定了精子阶段 Express ZP3调节的通道为分子克隆设定了阶段 以及随后的通道cDNA分析。