VITAMIN A AND REPRODUCTION

维生素 A 与生殖

• 批准号: 3326209
• 负责人: DAVID E ONG
• 金额: $ 13.67万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-30 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3326209
• 关键词: Sertoli cells; acyltransferase; cell free system; enzyme mechanism; epididymis; human tissue; laboratory rabbit; laboratory rat; protein purification; radioimmunoassay; receptor; retinoid binding proteins; retinoids; tissue /cell culture; transport proteins; vitamin metabolism

The broad long-term goal of this proposal is to understand the essential role that vitamin A plays in male reproductive function, seeking to understand the necessary movements and metabolism of vitamin A in the testis and epididymis. The specific research proposed seeks to clarify the roles of several specific transport or binding proteins for vitamin A in these organs. Particular emphasis is given to two previously undescribed retinoic acid-binding proteins present in the epididymis of the rat. It is proposed to purify and characterize these proteins including determination of sequence, binding specificity, and endogenous ligand. The distribution and cellular location of the proteins will be determined by preparation of antiserum that can be used for sensitive radioimmunoassay and immunolocalization. Other species will be examined for the presence of these proteins., The function of the proteins will be examined by studying their ability to deliver retinoic acid to developing sperm. Preliminary work has shown that Sertoli cells in primary culture will internalize retinol from its complex with retinol-binding protein, the plasma transport protein for vitamin A, in a receptor-mediated fashion. This process will be further characterized. The metabolism of the internalized retinol will be determined, with particular emphasis on esterification and the possible involvement of a novel enzyme activity, lecithinretinol acyl transferase. The possible release by the Sertoli cell of the internalized vitamin A to the medium will be examined. The plasma membrane receptor for RBP will be studied in cell-free systems, following the transfer of retinol from RBP to CRBP. Purification of the RBP receptor will be attempted in order to produce antiserum usable for radioimmunoassay and immunolocalization. Finally, the cell-specific location of cellular retinol-binding protein and cellular retinoic acid-binding protein in human reproductive tissue will be determined. With antiserum to human CRBP, localization studies in human testis and epididymis will be continued. Such studies will be extended to ovary and uterus, two tissues with abundant CRBP. Human CRABP will be purified and antiserum to CRABP obtained that will be used for radioimmunoassay and immunolocalization studies similar to those for CRBP.

该提案的广泛长期目标是了解基本内容 维生素 A 在男性生殖功能中的作用 了解维生素A在体内的必要运动和代谢 睾丸和附睾。拟议的具体研究旨在澄清 维生素 A 的几种特异性转运或结合蛋白的作用 在这些器官中。特别强调的是之前的两个 附睾中存在未描述的视黄酸结合蛋白 老鼠。建议纯化和表征这些蛋白质 包括序列、结合特异性和内源性的确定 配体。蛋白质的分布和细胞位置将是 通过制备可用于敏感人群的抗血清来测定 放射免疫测定和免疫定位。其他物种将接受检查 由于这些蛋白质的存在，蛋白质的功能将是 通过研究它们将视黄酸传递给发育中的能力来检查 精子。 初步工作表明，原代培养物中的支持细胞将 将视黄醇从其与视黄醇结合蛋白的复合物中内化， 维生素 A 的血浆转运蛋白，以受体介导的方式。 该过程将被进一步表征。的新陈代谢 将确定内化视黄醇，特别强调 酯化和可能涉及一种新的酶活性， 卵磷脂视黄醇酰基转移酶。塞尔托利可能释放 将检查细胞内化的维生素 A 到培养基中的情况。这 RBP 的质膜受体将在无细胞系统中进行研究， 视黄醇从 RBP 转移到 CRBP 后。纯化 将尝试 RBP 受体以产生可用于 放射免疫测定和免疫定位。 最后，细胞视黄醇结合蛋白的细胞特异性位置 和人类生殖组织中的细胞视黄酸结合蛋白 将被确定。人类CRBP抗血清的定位研究 人类的睾丸和附睾将继续。此类研究将 延伸至卵巢和子宫，这两种组织都含有丰富的CRBP。人类CRABP 将被纯化并获得抗CRABP的血清，其将用于 放射免疫测定和免疫定位研究类似于 CRBP。