MOLECULAR ANALYSIS OF THE SEA URCHIN SPERM RECEPTOR

海胆精子受体的分子分析

• 批准号: 2203027
• 负责人: KATHLEEN R FOLTZ
• 金额: $ 10.15万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 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 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2203027
• 关键词: Asteroidea; affinity chromatography; alternatives to animals in research; antireceptor antibody; cell membrane; complementary DNA; crosslink; cytoplasm; cytoskeletal proteins; egg /ovum; immunofluorescence technique; immunoprecipitation; microinjections; monoclonal antibody; peptides; phosphorylation; protein tyrosine kinase; receptor binding; receptor expression; recombinant proteins; sea urchins; sperm

Fertilization is the result of a set of well-choreographed events between the sperm and the egg. Successful gamete interactions result in the activation of the egg and initiate developmental pathways. Unsuccessful interactions, as a result of either sperm or egg dysfunction, are disastrous. Although much is known about the steps preceding binding and sperm-egg fusion, the mechanism of the binding event and the resulting signal transduction pathways that occur within the egg have been elusive. The recent identification and cloning of the sea urchin egg plasma membrane receptor for sperm now makes it possible to investigate these questions and to determine how the receptor functions at egg activation. This proposal describes experiments designed to address this structure/function question in the sea urchin, a well-characterized model system for fertilization and early development. The general question centers on the role that the receptor plays in transducing a signal in response to sperm binding, be it direct or indirect. Our preliminary data indicate that the receptor interacts with an egg cortical protein tyrosine kinase and may serve to organize the actin cytoskeletal rearrangements that occur at fertilization. The specific aims are: (i) To define the topology of the receptor in the membrane in order to establish which domains may interact with other egg proteins; (ii) To identify egg proteins that interact with the receptor; (iii) To test the functional role(s) of the cytoplasmic domain by injection of specific antibodies and peptides; and (iv) To initiate mutational analyses of the receptor, employing a heterologous expression system. These experiments will provide for a greater understanding of how sperm binding triggers the changes that occur in the egg, adding to our general knowledge of the basic processes of fertilization so important to our understanding of fertility and infertility.

受精是一系列精心设计的事件的结果 精子和卵子。成功的配子相互作用导致 激活卵子并启动发育途径。不成功 由于精子或卵子功能障碍而导致的相互作用 灾难性的。尽管人们对绑定之前的步骤了解很多， 精卵融合、结合事件的机制以及由此产生的结果 卵内发生的信号转导途径一直难以捉摸。 海胆卵血浆的最新鉴定与克隆 精子的膜受体现在使得研究这些成为可能 问题并确定受体在卵子激活时如何发挥作用。 该提案描述了旨在解决此问题的实验 海胆的结构/功能问题，一个良好表征的模型 受精和早期发育系统。一般问题 集中于受体在信号转导中所起的作用 对精子结合的反应，无论是直接的还是间接的。我们的初步数据 表明受体与卵皮质蛋白酪氨酸相互作用 激酶并可能有助于组织肌动蛋白细胞骨架重排 受精时发生的情况。具体目标是： (i) 定义 膜上受体的拓扑结构，以确定 结构域可能与其他卵蛋白相互作用； (ii) 识别鸡蛋 与受体相互作用的蛋白质； (iii) 测试功能 通过注射特异性抗体来发挥细胞质结构域的作用 肽； (iv) 启动受体的突变分析， 采用异源表达系统。这些实验将提供 更好地了解精子结合如何引发变化 发生在鸡蛋中，增加了我们对基本过程的常识 受精对于我们理解生育力非常重要 不孕症。