Photochemical Crosslinking Studies of Rat Epididymal Sperm Plasma Membrane

大鼠附睾精子质膜光化学交联研究

• 批准号: 8714741
• 负责人: Joe Hall
• 金额: $ 44.29万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8714741&HistoricalAwards=false
• 关键词: Photochemical; Crosslinking; Studies; Rat; Epididymal

The objective of this study is to provide a comprehensive evaluation of polypeptides exposed to the hydrophilic (peripheral proteins) or hydrophobic (integral proteins) region of the plasma membrane of rat testicular and epididymal spermatozoa. This will be done by reacting intact spermatozoa recovered from rete testis fluid and different regions of the epididymis with permeable, semi-permeable, or nonpermeable, radioactive chemical crosslinking reagents that are photosensitive. Following photolysis the plasma membrane will be purified by density gradient centrifugation, and the proteins will be analyzed by polyacrylamide gel electrophoresis and autoradiography. In other studies, nearest neighbor analysis of selected epididymal secreted proteins will be carried out. By identifying the major polypeptides that comprise the integral and peripheral proteins of the sperm plasma membrane as a function of epididymal transit, these studies may provide the groundwork for future investigations to test the functional significance of proteins that are added to, or removed from the sperm plasma membrane during development. The function of surface proteins during fertilization also represents an important direction for further research. Finally, these results may prove important to our understanding of the role secreted proteins play in the reproductive process.

这项研究的目的是提供一个全面的 评价暴露于亲水性（外周） 蛋白质）或疏水（整合蛋白质）区域 大鼠睾丸和附睾精子膜。 这将 通过使从睾丸网回收的完整精子 液体和附睾的不同区域具有渗透性， 半渗透性或非渗透性放射性化学品 光敏交联试剂。 以下 光分解质膜将被密度净化 梯度离心，蛋白质将通过 聚丙烯酰胺凝胶电泳和放射自显影。 在 其他研究，选择的附睾最近邻分析 分泌的蛋白质将被进行。 通过鉴定包含多肽的主要多肽， 精子质膜的整合蛋白和外周蛋白， 附睾运输的功能，这些研究可能提供 为今后的调查奠定基础， 蛋白质的重要性，添加到，或从 精子质膜在发育过程中。 的功能 受精过程中的表面蛋白也代表了一种 是进一步研究的重要方向。 最后，这些结果 可能对我们理解 蛋白质在生殖过程中发挥作用。