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（艾德）的功能， 附睾（EP）在精子成熟中的作用。本研究的目的是 检验雄激素或雌激素或两者都重要的假设 用于表达雄激素受体（AR）和雌激素受体（ER） 基因和/或蛋白质。表达的上调或下调可能取决于 根据区域、细胞类型以及艾德和EP的生长状态。 为了验证这一假设，我们建议表征mRNA和蛋白质， AR和ER在分子亚细胞水平上在不同区域的 成熟动物的艾德和EP（实验1），未成熟的生长动物（Exp. 2）、 有或没有雄激素替代的成熟去势动物，或 雌激素或两者（Exp. 4）。逆转录聚合酶链 反应技术将用于鉴定mRNA转录本，并在原位 杂交将用于在亚细胞水平定位mRNA。 免疫组织化学和Western印迹技术将用于 鉴定AR和ER蛋白。mRNA和蛋白质的变化将与 与那些在结构形态测量学和激素谱（睾酮， 雌激素和促黄体激素）。 而前两个实验的结果将提供半定量的 AR和ER的基因/蛋白表达数据作为区域的函数 和发展，后两个实验将提供 雄激素和雌激素之间差异显著性的半定量数据 雌激素以及循环因素和睾丸因素之间的关系， 调节AR和ER基因/蛋白表达。总体而言，结果将 提供了关于功能性调节艾德和EP的见解， 在精子成熟中起重要作用，因此对男性生育力也很重要。

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.