MALE REPRODUCTIVE PHYSIOLOGY

本研究的长期目标是阐明 负责精子发育的机制 哺乳动物附睾的生殖能力。 作为精子 通过附睾，它们依次暴露于 由分泌物产生的不同附睾环境， 附睾细胞的吸收。 工作假设是， 精子所必需的连续质膜的成熟- 卵的相互作用依赖于与 尿道控制的附睾分泌蛋白。 以下 已经开发了以下程序：1）在体外维持 附睾外植体中所有细胞之间的相互作用 区域及其三维结构关系， 保存，和2）分离和培养活的上皮细胞， 附睾管周细胞 该提案的具体目标是：1）研究蛋白质 以及附睾细胞的糖蛋白合成和分泌。 这 将被研究后，标记糖和氨基 酸，通过SDS-PAGE电泳和免疫沉淀; 2） 研究蛋白质和糖蛋白合成的激素控制 和分泌; 3）测定分泌蛋白的结合， 糖蛋白对附睾精子的作用及其对 精子-卵子结合，然后对精子可受精性; 4） 确定附睾分泌的区域差异， 分泌-精子相互作用：5）研究附睾分泌 能够修饰精子表面的糖苷酶; 6）研究 糖苷酶诱导的结构和功能的变化 分泌的附睾和精子糖蛋白; 7）研究附睾 精子分泌及分泌-精子相互作用 基因突变与正常的精子生产，精子形态， 精子活力和低生育力 兔子、老鼠和老鼠 被利用 这些研究应该提供重要的见解， 男性生育问题。

The long term objective of this research is the elucidation of the mechanisms which are responsible for the development of sperm fertilizing capacity in the mammalian epididymis. As spermatozoa pass through the epididymis, they are sequentially exposed to varying epididymal environments created by the secretion and absorption of epididymal cells. The working hypothesis is that the maturation of the sequential plasma membrane necessary for sperm- egg interaction is dependent upon the sequential interaction with hormonally controlled epididymal secretory proteins. The following procedures have been developed: 1) maintenance in vitro of epididymal explants where the interaction between all the cells of a region and their 3-dimensional structural relationship are preserved, and 2) isolation and culture of viable epithelial and peritubular epididymal cells. The specific objectives of this proposal are 1) to study protein and glycoprotein synthesis and secretion by epididymal cells. This will be studied, after incorporation of labeled sugar and amino acids, by SDS-PAGE electrophoresis and immunoprecipitation; 2) to study the hormonal control of protein and glycoprotein synthesis and secretion; 3) to determine the binding of secreted proteins and glycoprotein to epididymal spermatozoa and their effect first on sperm-egg binding and then on sperm fertilizing ability; 4) to determine the regional differences in epididymal secretion and secretion-sperm interactions: 5) to study the epididymal secretion of glycosidases which can modify the sperm surface; 6) to study glycosidase inhibitor-induced changes in structure and function of secreted epididymal and sperm glycoproteins; 7) to study epididymal secretion and secretion-sperm interaction in mice bearing single gene mutations with normal sperm production, sperm morphology, sperm motility, and low fertility. Rabbits, rats, and mice will be used. These studies should provide important insights into the problem of male fertility.