REGULATION OF EFFERENT DUCTULES EPIDIDYMIS

附睾传出小管的调节

• 批准号: 6352965
• 负责人: HARI Om GOYAL
• 金额: $ 11万
• 依托单位: TUSKEGEE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6352965
• 关键词: androgen receptor; electron microscopy; epididymis; estrogen receptors; gene expression; goats; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; light microscopy; male castration; male reproductive system; messenger RNA; molecular cloning; northern blottings; polymerase chain reaction; protein biosynthesis; receptor binding; southern blotting; spermatogenesis; statistics /biometry; testis; testosterone; western blottings

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.

该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）的功能和 附睾（EP）参与精子的成熟。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 用于雄激素受体 (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 取决于 ED 和 EP 的区域、细胞类型以及生长状态。 为了检验这个假设，我们建议表征 mRNA 和蛋白质 不同区域的分子亚细胞水平的 AR 和 ER 成熟动物（实验 1）、未成熟生长动物（实验 2）的 ED 和 EP， 成熟的阉割动物，用或不用雄激素替代或 雌激素或两者（实验 4）。逆转录酶聚合酶链 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。 免疫组织化学和蛋白质印迹技术将用于 识别 AR 和 ER 蛋白。 mRNA和蛋白质的变化会相关 与结构形态测量和荷尔蒙概况（睾酮， 雌激素和黄体生成素）。 而前两个实验的结果将提供半定量 AR 和 ER 的基因/蛋白质表达数据作为该区域的函数 和发展，后两个实验将提供 雄激素和雄激素之间差异显着性的半定量数据 雌激素以及循环因子和睾丸因子之间 调节 AR 和 ER 基因/蛋白表达。总体而言，结果将 提供有关 ED 和 EP 监管的见解，这些监管功能在功能上 对于精子成熟非常重要，因此对于男性生育能力也很重要。