EPIDIDYMAL IMMOBILIN--STRUCTURE, FUNCTION, BIOGENESIS

附睾固定素——结构、功能、生物发生

• 批准号: 3448393
• 负责人: Norka D. Ruiz Bravo
• 金额: $ 4.58万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-01 至 1989-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3448393
• 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.

固定素，一种新的高相对分子质量的糖蛋白(粘蛋白)，在 大鼠附睾液抑制大鼠运动的启动 精子。固定素似乎通过其功能抑制精子的活力 粘弹性，而不是它的生物化学特性。一批 关于固定化的重要生理和生化问题 仍未得到回复： 1.固定素的结构与其抑制精子的关系 能动性。 固定素及其粘弹性与其相互作用的可能相关性 将检查抑制精子运动的能力。Immobilin将成为 纯品及其氨基酸和碳水化合物组成 下定决心。固定素将进行化学和酶修饰，以及 每一种修饰对固定素生物活性的影响将是 下定决心。 2.固定化蛋白的生物发生。 将在兔体内产生针对固定素的多克隆抗体，并将其用作 探头： A)固定素的合成部位。组织和细胞的特异性 固定素的合成将通过免疫印迹和生化两种方法进行检测。 和均质组织的免疫沉淀，在形态上，通过 免疫标记组织切片。 B)对固定素生物合成的调控。一旦组织和细胞 固定素生物合成的特异性已被确定，个体发育 本课程将研究固定素的生物合成。适当的匀浆 来自胎儿、新生、青春期和成年大鼠的器官将被免疫 已经检查过是否有固定素。此外，无论是最初出现的 附睾尾部管腔内的固定素蛋白是由于 随之而来的是功能性(可翻译的)固定素mRNA的出现 通过体外翻译和免疫沉淀间接确定 新合成的固定素。 固定素生物合成的可能的激素调节将是 活体研究。荷尔蒙的状态会受到 垂体切除、去势、输出管结扎和雄激素 替代疗法。雄激素是否通过增加 体外实验将检测功能性Immobilin mRNA的水平。 新合成的固定素的翻译和免疫沉淀。