WATER AND ELECTROLYTE MOVEMENT IN RENAL TUBULES

这项提议的总体目标是建立有意义的 哺乳动物肾单位的结构-功能关系。 将进行三条主要调查路线，其中两条 代表正在进行的研究的自然进展，这将是 通过开发新的 方法和新的协作关系。三位一体 研究领域包括异噬和自噬的检验。 在肾小管上皮细胞的定位和作用机制 质子和碳酸氢盐在收集管道中的传输，以及 继续对肾脏进行形态观察。更多 具体地说： 1.自噬和异噬的过程将是 在哺乳动物近端小管中进行评估以更好地表征 亚细胞器、蛋白质和相关物质 分子物质会发生降解。特别强调将 放在测定组织蛋白酶B基因表达上作为一种 更好地理解这种机制的手段 控制自噬空泡-溶酶体中组织蛋白酶B的水平 车厢。 2.质子和亚细胞事件涉及的位置和亚细胞事件 将评估碳酸氢盐在哺乳动物体内的运输情况 通过检查管道的结构和功能来收集风管 主细胞和嵌合细胞。严重依赖将会 被放在运输蛋白和离子的鉴定上 通过使用高度特异的抗体探针和 免疫细胞化学和光亲和标记技术。 3.交感神经纤维在脑内的分布 哺乳动物肾脏与神经效应物质的关系 肾小管和肾小管上的连接和α、β肾上腺素受体 将对血管系统进行检查。我们将特别强调 α和β肾上腺素能受体的识别贯穿始终 超微结构水平上的肾实质 针对受体的特异性抗体和通过 放射性标记肾上腺素能的放射自显影定位 使用光亲和标记的拮抗剂。 这些调查将需要应用各种 生理、生化和形态技术，包括 光学显微镜、透射电子显微镜和扫描电子显微镜， 体外光学和电子显微镜放射自显影 离体肾小管微灌流、超微酶 分析和免疫细胞化学。

The overall objective of this proposal is to establish meaningful structural-functional relationships in the mammalian nephron. Three major lines of inquiry will be pursued, two of which represent natural progressions of ongoing research that will be expanded and intensified through the development of new methodologies and new collaborative relationships. The three areas of inquiry include examination of heterophagy and autophagy in the renal tubule, delineation of the sites and mechanisms of proton and bicarbonate transport in the collecting duct, and continued morphological exploration of the kidney. More specifically: 1. The processes of autophagy and heterophagy will be evaluated in the mammalian proximal tubule to better characterize the manner in which subcellular organelles, proteins and related molecular substances undergo degradation. Special emphasis will be placed on the measurement of cathepsin B mRNA expression as a means of obtaining a better understanding of the mechanism that controls cathepsin B levels in the autophagic vacuole-lysosomal compartment. 2. The sites and subcellular events involved in proton and bicarbonate transport will be evaluated along the mammalian collecting duct by examining the structure and function of the principal cells and the intercalated cells. Heavy reliance will be placed on identification of transport proteins and ion channels through the use of highly specific antibody probes and immunocytochemical and photoaffinity labelling techniques. 3. The distribution of the sympathetic nerve fibers in the mammalian kidney and the relationship between neuroeffector junctions and alpha- and beta-adrenoceptors on tubules and vasculature will be examined. Special emphasis will be placed on the identification of alpha- and beta-adrenoceptors throughout the renal parenchyma at the ultrastructural level with the use of specific antibodies directed against the receptors and by autoradiographic localization of radiolabelled adrenergic antagonists using photoaffinity labelling. These investigations will require the application of a variety of physiologic, biochemical and morphologic techniques including light microscopy, transmission and scanning electron microscopy, light and electron microscopic autoradiography, in vitro microperfusion of isolated kidney tubules, ultramicroenzyme analysis and immunocytochemistry.