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: 8242803
• 负责人: Michael B Butterworth
• 金额: $ 24.41万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8242803
• 关键词: Address; Apical; Bartter Disease; Biochemical; Biological; Biological Assay; Biotinylation; Cell Line; Cell physiology; Cells; Cellular biology; Chimeric Proteins; Collaborations; Cyclic AMP; Data; Defect; Disease; Distal; Docking; Duct (organ) structure; Electrons; Epithelial; Epithelium; Equilibrium; Event; Feedback; Guanosine Triphosphate; Hormonal; Hypertension; Image; Institution; Ion Channel; Ions; KCNJ1 gene; Kidney; Label; Life; Location; Mediating; Membrane; Microscopic; Modeling; Movement; Mus; Nephrogenic Diabetes Insipidus; Nephrons; Pathway interactions; Physiological; Play; Population; Potassium Channel; Principal Investigator; Process; Proteins; Pseudohypoaldosteronism; 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; renal epithelium; trafficking; uptake; water channel

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.

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、假性醛固酮增多症 1 型和利德尔综合征。盐和水在皮质集合管中的运输 肾脏的 (CCD) 通过受激素反馈严格控制的特定通道介导 机制。拟议的研究将通过以下方式定义和表征新的渠道调节机制： 囊泡运输和回收。这些研究旨在调查三种最常见的情况是否 CCD 顶膜上的转运蛋白，即上皮钠通道、钾通道和水通道 由囊泡运输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 对于这些渠道的规范回收非常重要。多学科方法将首先确定亚细胞 通过免疫细胞化学标记以及荧光和电子的囊泡运输区室 显微成像。接下来，将验证这些囊泡室中通道的定位 生物化学上。贩运事件的动态将通过活细胞成像和 电生理技术。最后，规范这些渠道回收的机制将 予以阐明。通过研究 Rab 蛋白在调节通道循环中的作用， 将确定参与渠道贩运的组成部分。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。这不仅有助于理解 涉及盐和水平衡的重要稳态过程，但更重要的是，这项工作将 深入了解该法规中与渠道监管不当相关的潜在缺陷，从而导致 处于病理生理疾病状态。

项目成果

MicroRNAs and the regulation of aldosterone signaling in the kidney.

MicroRNA 和肾脏中醛固酮信号传导的调节。

• DOI: 10.1152/ajpcell.00026.2015
• 发表时间: 2015-04-01
• 期刊: American journal of physiology. Cell physiology
• 作者: M. Butterworth

Role of microRNAs in aldosterone signaling.

microRNA 在醛固酮信号传导中的作用。

• 发表时间: 2018
• 影响因子: 3.2
• 作者: Butterworth; Michael B
• 通讯作者: Michael B