REGULATION OF EFFERENT DUCTULES EPIDIDYMIS

这项研究的长期目标是了解雄激素的作用 和/或雌激素调节传出小管(ED)和 精子成熟过程中的附睾部。本研究的目的是 检验雄激素或雌激素或两者都很重要的假设 雄激素受体(AR)和雌激素受体(ER)的表达 基因和/或蛋白质。表达的上调或下调可能取决于 根据ED和EP的区域、细胞类型和生长状态。 为了验证这一假设，我们建议对mRNAs和蛋白质进行表征 不同区域AR和ER在分子亚细胞水平的表达 成熟动物的ED和EP(Exp.1)、幼龄生长动物(Exp.2), 成熟的去势动物，无论是否替代雄激素或 雌激素或两者兼而有之(实验4)。逆转录酶聚合酶链 反应技术将被用来鉴定mRNA转录本，并原位 杂交将被用来在亚细胞水平上定位mRNA。 免疫组织化学和免疫印迹技术将用于 识别AR和ER蛋白。MRNAs和蛋白质的变化将与 与那些结构形态和激素谱(睾酮， 雌激素和黄体生成素)。 鉴于前两个实验的结果将提供半定量的 AR和ER基因/蛋白表达随区域变化的数据 和开发，后两个实验的结果将提供 雄激素与雄激素差异显著的半定量数据 雌激素以及循环因子和睾丸因子之间的关系 调节AR和ER基因/蛋白的表达。总体而言，结果将 就教育署和环境保护局的监管提供见解 对精子成熟很重要，因此对男性生育能力也很重要。

The long term goal of the study is to understand the role of androgens and/or estrogens in regulating functions of the efferent ductules (ED) and epididymis (EP) in sperm maturation. The aim of the present study is to test the hypothesis that androgens, or estrogens, or both are important for expression of the androgen receptor (AR) and estrogen receptor (ER) genes and/or proteins. The up-or down-regulation of expression may depend upon the region, the cell type, and the state of growth of the ED and EP. To test the hypothesis we propose to characterize mRNAs and proteins for AR and ER at the molecular subcellular levels in different regions of the ED and EP of mature animals (Exp. 1), immature growing animals (Exp. 2), mature castrated animals with or without substitution of androgens or estrogens or both (Exp. 4). The reverse transcriptase polymerase chain reaction technique will be used to identify mRNA transcripts, and in situ hybridization will be used to localize mRNA at the subcellular level. Immunohistochemical and Western blotting techniques will be used to identify AR and ER proteins. Changes in mRNAs and proteins will be related with those in structural morphometry and hormonal profiles (testosterone, estrogen, and luteinizing hormone). Whereas results of the first two experiments will provide semiquantitative data on gene/protein expression for AR and ER as a function of the region and development, those from the latter two experiments will provide semiquantitative data on differential significance between androgens and estrogens as well as between circulating factors and testicular factors in regulating AR and ER gene/protein expression. Overall, results will provide insights on regulation of the ED and EP which are functionally important in sperm maturation and, thus, in male fertility.