Photochemical Crosslinking Studies of Rat Epididymal Sperm Plasma Membrane

本研究的目的是对暴露在大鼠睾丸和附睾精子质膜亲水性（外周蛋白）或疏水性（整体蛋白）区域的多肽进行综合评价。这将通过与可透性、半透性或不可透性的放射性化学交联试剂反应从睾丸精管液和附睾不同区域恢复的完整精子来完成。光解后，质膜将通过密度梯度离心纯化，蛋白质将通过聚丙烯酰胺凝胶电泳和放射自显影进行分析。在其他研究中，将对选定的附睾分泌蛋白进行最近邻分析。通过鉴定构成精子质膜整体蛋白和外周蛋白的主要多肽在附睾转运过程中的作用，这些研究可能为未来的研究奠定基础，以测试在发育过程中添加到精子质膜或从精子质膜中移除的蛋白的功能意义。表面蛋白在受精过程中的作用也是今后研究的一个重要方向。最后，这些结果可能对我们理解分泌蛋白在生殖过程中所起的作用很重要。

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.