EPIDIDYMAL IMMOBILIN--STRUCTURE, FUNCTION, BIOGENESIS

附睾固定素——结构、功能、生物发生

• 批准号: 3448391
• 负责人: Norka D. Ruiz Bravo
• 金额: $ 5.04万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3448391
• 关键词: androgens; animal age group; antibody; autoradiography; carbohydrate structure; epididymis; gel electrophoresis; genetic translation; growth /development; hormone regulation /control mechanism; hormone therapy; hypophysectomy; immunologic assay /test; male castration; messenger RNA; sperm motility; spermicide; viscosity

Immobilin, a novel glycoprotein (mucin) of high molecular weight found in rat cauda epididymal fluid, inhibits the initiation of motility of rat sperm. Immobilin appears to inhibit sperm motility by virtue of its viscoelasticity rather than by its biochemical properties. A number of physiologically and biochemically important questions about immobilin remain unanswered: 1. Relationship of immobilin's structure to its inhibition of sperm motility. The possible correlation between immobilin, its viscoelasticity and its ability to inhibit sperm motility will be examined. Immobilin will be purified to homogeneity and its amino acid and carbohydrate composition determined. Immobilin will be chemically and enzymatically modified, and the effects of each modification on immobilin's biological activity will be determined. 2. Biogenesis of immobilin. Polyclonal antibodies to immobilin will be generated in rabbits and used as probes: a) Site of synthesis of immobilin. The tissue and cell specificity of immobilin synthesis will be examined both biochemically, by immunoblotting and immunoprecipitation of homogenized tissues, and morphologically, by immunolabeling tissue sections. b) Regulation of immobilin biosynthesis. Once the tissue and cell specificity of immobilin biosynthesis have been determined, the ontogeny of immobilin biosynthesis will be examined. Homogenates of the appropriate organ from fetal, neonatal, pubertal and adult rats will be immunologically screened for immobilin. In addition, whether the initial appearance of immobilin protein in the lumen of the cauda epididymis is due to the concomitant appearance of functional (translatable) immobilin mRNA will be indirectly determined by in vitro translations and immunoprecipitation of newly synthesized immobilin. The possible hormonal regulation of immobilin biosynthesis will be investigated in vivo. The hormonal status will be manipulated by hypophysectomy, castration, efferent duct ligation, and androgen replacement therapy. Whether androgens exert their effect by increasing the levels of functional immobilin mRNA will be examined by in vitro translation and immunoprecipitation of newly synthesized immobilin.

Immobilin是一种新的高分子量的糖蛋白（粘蛋白）， 大鼠附睾尾液，抑制大鼠运动的启动 精子 固定蛋白似乎通过其在精子中的作用抑制精子活力。 粘弹性而不是其生化性质。 一些 关于固定蛋白的生理学和生物化学的重要问题 仍未得到答复： 1. immobilin的结构与其精子抑制作用的关系 能动性 immobilin及其黏弹性与其黏弹性之间可能存在的相关性 将检查抑制精子运动的能力。 Immobilin将 纯化至均一，其氨基酸和碳水化合物组成 测定 固定蛋白将被化学和酶修饰， 每种修饰对固定素生物活性的影响将是 测定 2. immobilin的生物发生。 将在兔中产生针对immobilin的多克隆抗体，并用作 探头： a）固定素的合成位点。 组织和细胞特异性 将通过生物化学、免疫印迹 和免疫沉淀的匀浆组织，并在形态学上， 免疫标记组织切片。 B）固定素生物合成的调节。 一旦组织和细胞 已经确定了immobilin生物合成的特异性， 将检查固定素生物合成。 适当的匀浆 将对来自胎儿、新生、青春期和成年大鼠的器官进行免疫学检查。 筛选固定素。 此外，无论最初出现的 附睾尾管腔中的immobilin蛋白是由于 伴随出现的功能性（可翻译的）immobilin mRNA将是 通过体外翻译和免疫沉淀间接测定 新合成的immobilin 可能的激素调节immobilin生物合成将是 在体内研究。 荷尔蒙的状态会被 垂体切除术、去势术、输出管结扎术和雄激素 替代疗法 雄激素是否通过增加 功能性固定蛋白mRNA的水平将通过体外 新合成的固定素的翻译和免疫沉淀。