EnaC regulation in the kidney by vesicle trafficking and recycling

肾脏离子和水分调节的缺陷与许多疾病有关，包括， 肾源性尿崩症、高血压、巴特综合征、吉特曼综合征、假性醛固酮减少症 1型和利德尔综合征。大脑皮层集合管中的盐分和水分运输 肾脏(Ccd)是通过特定的受激素反馈严格控制的通道来调节的。 机械装置。拟议的研究将通过以下方式定义和描述新的渠道调节机制 囊泡贩运和回收。这些研究旨在调查三种最常见的 上皮钠、钾和水通道是位于CCD顶膜的转运体。 由囊泡贩运共同定位和共同调节。使用小鼠CCD细胞系的研究将定义 这些贩运事件的潜在生理调节。研究将确定特定的细胞机制 在这些渠道的规范回收方面非常重要。一个多学科的方法将首先确定亚细胞 应用免疫细胞化学标记及荧光和电子技术研究囊泡转运室 显微成像。接下来，将验证通道在这些囊泡隔室中的定位 生化方面的。贩运活动的动态将使用活细胞成像和 电生理技术。最后，规范这些渠道循环利用的机制将 将被澄清。通过研究Rab蛋白在调节通道循环中的作用 将确定涉及渠道贩运的组件。这些基础细胞生物学研究将进一步 了解肾脏中水和离子通道的调节。这不仅有助于理解 涉及盐和水平衡的基本动态平衡过程，但更重要的是，这项工作将 深入了解此法规中与渠道失控相关的潜在缺陷 处于病理生理疾病状态。

Defects in ion and water regulation in the kidney have been associated with numerous diseases including, nephrogenic diabetes insipidus, hypertension, Bartter syndrome, Gitelman syndrome, pseudohypoaldosteronism type 1 and Liddle syndrome. Salt and water transport in the cortical collecting duct (CCD) of the kidney is mediated through specific channels which are tightly controlled by hormonal feedback mechanisms. The proposed studies will define and characterize novel mechanisms of channel regulation by vesicle trafficking and recycling. The studies aim to investigate whether three of the most common transporters at the CCD apical membranes, namely the epithelial sodium, potassium and water channels are co-localized and co-regulated by vesicle trafficking. Studies using a mouse CCD cell line will define the underlying physiological regulation of these trafficking events. Studies will identify specific cell machinery important in the regulated recycling of these channels. A multidisciplinary approach will first identify the subcellular vesicle trafficking compartments by immuno-cytochemical labeling and fluorescent and electron microscopic imaging. Next, the localization of channels in these vesicle compartments will be verified biochemically. The dynamics of the trafficking events will be determined using live-cell imaging and electrophysiological techniques. Finally, the mechanisms which regulate the recycling of these channels will be elucidated. By investigating the role of Rab-proteins in regulated channel recycling the essential cellular components involved in channel trafficking will be determined. These basic cell biological studies will further an understanding of water and ion channel regulation in the kidney. This will help not only to understand the essential homeostatic processes involved in salt and water balance, but more importantly, the work will provide insights into underlying defects in this regulation associated with channel misregulation which result in pathophysiological disease states.