ACIDIFICATION MECHANISMS IN THE EPIDIDYMIS/VAS DEFERENS

附睾/输精管的酸化机制

• 批准号: 6296454
• 负责人: SYLVIE BRETON
• 金额: $ 26.21万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6296454
• 关键词: acid base balance; adrenergic agents; aldosterone; antiadrenergic agents; apical membrane; carbonate dehydratase; cytoskeletal proteins; epididymis; guanine nucleotide binding protein; hormone inhibitor; immunocytochemistry; in situ hybridization; ion transport; laboratory rat; membrane transport proteins; northern blottings; testosterone; vesicle /vacuole

The acidic lumenal pH in the epididymis and vas deferens is largely generated by a bafilomycin-sensitive H-ATPase on the plasma membrane and intracellular vesicles of a population of specialized epithelial cells that resemble kidney collecting duct intercalated cells in many respects. This acidic pH plays a critical role in sperm maturation and helps maintain sperm in a quiescent state until ejaculation: defective acidification may impair reproductive function in pathologic conditions, while intervention to manipulate acidification male be used to control male fertility. The aim of this proposal is to develop a thorough understanding of the regulatory mechanisms involved in this acidification process. The central hypothesis behind the proposed studies is that the differentiation and functional activity of these proton pumping cells is regulated physiologically, that vesicle recycling is involved in controlling the plasma membrane content of H-ATPase, and that transepithelia proton secretion involves parallel ion transporting proteins. A unique aspect of the proposed studies is the use of a multidisciplinary approach that will benefit greatly from the combined expertise of participants in the other projects of this PPG application. Cell and molecular biology, and immunocytochemistry will be combined with measurements of transepithelial ion flux using ion- specific vibrating probe technology, and single-cell spectrofluorimetric analysis, to identity active transporters in the intact epithelium. A wide variety of antibody and cDNA reagents are available for these studies, and a carbonic anhydrase II-deficient mouse model with potentially defective luminal acidification will also be examined. Specific aims are: 1) To define the pathways of proton secretion in normal epididymis and vas deferens, and the parallel transport proteins (e.g., Na/H and anion exchangers; Na-HC03 and NaK/2C1 co-transporters) that are involved: 2) To define the cell biological and physiological regulatory mechanism(s) of H-ATPase recycling in the epididymis and vas deferens, including the role of the cytoskeleton, GTP-binding proteins, acid-base balance and hormones: 3) To follow the development of PP-rich cells and acidification capacity in excurrent ducts during postnatal maturation. Normal animals and carbonic- anhydrase-deficient mice will be used for these studies. These studies will provide important information on the poorly understood process of reproductive tract acidification, as well as exploiting a novel epithelial model to dissect and distinct intracellular vesicle recycling pathway.

附睾和VAS延期中的酸性腔内pH很大 由血浆上的Bafilomycin敏感HATPase产生 专业人群的膜和细胞内囊泡 上皮细胞，类似于肾脏收集管道插入细胞的细胞 许多方面。这种酸性pH在精子成熟中起关键作用 并有助于将精子保持在静止状态，直到射精为止： 缺陷酸化可能会损害病理学的生殖功能 条件，在处理酸化男性的干预措施时 控制男性生育能力。 该提议的目的是开发 对此涉及的监管机制的透彻理解 酸化过程。提议背后的中心假设 研究是这些质子的分化和功能活性 在生理上调节泵送细胞，囊泡回收是 参与控制H ATPase的质膜含量，并参与 那个transepithelia质子分泌涉及平行离子运输 蛋白质。 拟议研究的一个独特方面是使用 多学科的方法将从合计中受益匪浅 该PPG应用程序其他项目的参与者的专业知识。 细胞和分子生物学以及免疫细胞化学将是 结合使用离子 - 特定的振动技术和单细胞光谱 分析，在完整上皮中的身份活性转运蛋白。 宽 这些研究都可以使用多种抗体和cDNA试剂 以及具有潜在的碳酸酐酶II缺陷小鼠模型 还将检查腔酸缺陷。 具体目标 是：1）定义正常质子分泌的途径 附睾和VAS延迟，以及平行的转运蛋白（例如， NA/H和阴离子交换器； NA-HC03和NAK/2C1共发器） 所涉及的：2）定义细胞生物学和生理 Epidymis和H-ATPase回收的调节机制 VAS延期，包括细胞骨架的作用，GTP结合 蛋白质，酸碱平衡和激素：3）遵循 富含PP的细胞的发展和酸化能力 产后成熟期间的管道。 正常动物和碳 这些研究将使用缺血酶缺陷小鼠。 这些研究 将提供有关较了解过程的重要信息 生殖道酸化，并利用新型上皮 解剖和不同细胞内囊泡回收途径的模型。