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REGULATION OF RENAL NA/K-ATPASE ACTIVITY

肾 NA/K-ATP 酶活性的调节

基本信息

批准号：
3152799
负责人：
BARBARA M RAYSON
金额：
$ 9.31万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-09-01 至 1988-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3152799
关键词：
adenosinetriphosphatase cell membrane electrolyte balance fluorescence spectrometry histochemistry /cytochemistry hormone regulation /control mechanism intracellular intracellular membranes ion transport nuclear magnetic resonance spectroscopy potassium renal tubular transport

项目摘要

The number of Na/K-ATPase sites in the plasma membrane of Hela cells have previously been demonstrated to be regulated by changes in intracellular electrolyte levels. Preliminary evidence suggests that a similar regulatory mechanism is demonstrable in a superfused suspension of outer medullary kidney tubule segments, as described in this proposal. The superfused kidney tubular preparation provides an ideal system in which to study this mechanism. Intracellular electrolyte levels can be manipulated through precisely regulated changes in extracellular composition and measured directly using techniques which have been established in this particular preparation. Intracellular Na+ levels will be measured using NMR techniques and intracellular Ca2+ levels will be measured using the fluorescent indicator 'Quin 2'. In addition, it is proposed to establish the NMR technique of intracellular K+ level measurement. Intracellular K+ levels will also be determined in the characterization of the response described. In vivo, the changes required would be very complex and difficult to interpret. Finally, the yield of cells, characterized by high Na/K-ATPase concentrations, will provide the material necessary for the investigation of the cellular mechanisms involved. The first goal of the proposed project is to characterize the changes in Na/K-ATPase activity observed after chronic changes in intracellular electrolyte levels. This characterization will contribute to the definition of steroidal cellular actions, which are believed to implicate changes in intracellular electrolyte levels. The second goal is to define the ion species responsible for the changes in Na/K-ATPase activity observed. Finally, the third goal is to investigate the possible mechanisms involved in the regulation of Na/K-ATPase activity by chronic changes in intracellular electrolyte levels. (i) The possible association between changes in Na/K-ATPase activity and changes in plasma membrane area will be investigated to determine whether the response observed is a reflection of an increased concentration of transporting sites. (ii) To test whether changes in intracellular electrolyte levels unmask latent Na/K-ATPase sites. (iii) To determine whether changes in either enzyme synthetic or degradative rates are involved, and if so (iv) to examine possible mechanisms implicated in such changes.

Hela 细胞质膜中 Na/K-ATPase 位点的数量先前已被证明受细胞内变化的调节电解质水平。初步证据表明，类似的调节机制可以在外部的超融合悬浮中得到证明髓质肾小管段，如本提案中所述。灌注肾管制剂提供了一个理想的系统，其中来研究这个机制。细胞内电解质水平可通过精确调节细胞外的变化来操纵成分并直接使用已被证实的技术进行测量在此特定准备中建立。细胞内 Na+ 水平将使用 NMR 技术测量细胞内 Ca2+ 水平使用荧光指示剂“Quin 2”进行测量。此外，它是提出建立细胞内K+水平的NMR技术测量。细胞内 K+ 水平也将在描述的响应的特征。在体内，需要改变将非常复杂且难以解释。最后，产量为细胞以高 Na/K-ATPase 浓度为特征，将提供研究细胞机制所需的材料涉及。拟议项目的第一个目标是描述变化的特征细胞内慢性变化后观察到的Na/K-ATP酶活性电解质水平。这种特征将有助于类固醇细胞作用的定义，据信涉及细胞内电解质水平的变化。第二个目标是定义导致变化的离子种类观察到 Na/K-ATP 酶活性。最后，第三个目标是研究可能涉及的机制通过慢性变化调节Na/K-ATP酶活性细胞内电解质水平。 (i) 之间可能存在的关联 Na/K-ATP酶活性的变化和质膜面积的变化将进行调查以确定观察到的响应是否反映了运输地点更加集中。 (ii) 测试是否细胞内电解质水平的变化揭示了潜在的Na/K-ATP酶网站。 (iii) 确定酶合成或酶的变化是否发生变化涉及降解率，如果是的话 (iv) 检查可能的这些变化所涉及的机制。