EnaC regulation in the kidney by vesicle trafficking and recycling

通过囊泡运输和再循环调节肾脏中的 EnaC

• 批准号: 7995590
• 负责人: Michael B Butterworth
• 金额: $ 5.11万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-16 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7995590
• 关键词: Address; Apical; Bartter Disease; Biochemical; Biological; Biological Assay; Biotinylation; Cell Line; Cells; Cellular biology; Chimeric Proteins; Collaborations; Cyclic AMP; Data; Defect; Disease; Distal; Docking; Duct (organ) structure; Electrons; Epithelial; Epithelium; Equilibrium; Event; Feedback; Hormonal; Hypertension; Hypoaldosteronism; Image; Institution; Ion Channel; Ions; KCNJ1 gene; Kidney; Label; Life; Location; Mediating; Membrane; Microscopic; Modeling; Movement; Mus; Nephrogenic Diabetes Insipidus; Nephrons; Pathway interactions; Physiological; Physiology; Play; Population; Potassium; Potassium Channel; Principal Investigator; Process; Proteins; Recycling; Regulation; Research Personnel; Role; Sodium; Sodium Channel; Sodium Chloride; Surface; Syndrome; Techniques; Vasopressins; Vesicle; Water; Work; absorption; apical membrane; aquaporin-2; cellular imaging; collecting tubule structure; density; driving force; epithelial Na+ channel; insight; interdisciplinary approach; knock-down; novel; overexpression; preference; programs; renal epithelium; trafficking; uptake; water channel

DESCRIPTION (provided by applicant): Defects in ion and water regulation in the kidney have been associated with numerous diseases including, nephrogenic diabetes insipidus, hypertension, Bartter syndrome, Gitelman syndrome, pseudo-hypoaldosteronism 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 sub-cellular 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.

描述（由申请人提供）： 肾脏中离子和水调节的缺陷与许多疾病有关，包括肾性尿崩症、高血压、Bartter 综合征、Gitelman 综合征、1 型假性醛固酮减少症和 Liddle 综合征。肾脏皮质集合管（CCD）中盐和水的运输是通过特定通道介导的，这些通道受到激素反馈机制的严格控制。拟议的研究将定义和表征通过囊泡运输和回收进行通道调节的新机制。这些研究旨在调查 CCD 顶膜上三种最常见的转运蛋白，即上皮钠通道、钾通道和水通道是否共同定位并受到囊泡运输的共同调节。使用小鼠 CCD 细胞系的研究将确定这些贩运事件的潜在生理调节。研究将确定在这些通道的调节回收中重要的特定细胞机制。多学科方法将首先通过免疫细胞化学标记以及荧光和电子显微镜成像来识别亚细胞囊泡运输区室。接下来，将对这些囊泡区室中通道的定位进行生化验证。贩运事件的动态将使用活细胞成像和电生理学技术来确定。最后，将阐明调节这些通道循环利用的机制。通过研究 Rab 蛋白在调节通道再循环中的作用，将确定参与通道运输的重要细胞成分。这些基础细胞生物学研究将进一步了解肾脏中的水和离子通道调节。这不仅有助于了解盐和水平衡所涉及的基本稳态过程，更重要的是，这项工作将深入了解与通道失调相关的潜在缺陷，从而导致病理生理疾病状态。

项目成果

USP10: the nexus between nexin and vasopressin.

USP10：连接蛋白和加压素之间的联系。

• 发表时间: 2008-10
• 作者: Butterworth, Michael B;Johnson, John P
• 通讯作者: Johnson, John P