Endothelial cell potassium channels

内皮细胞钾通道

• 批准号: 9363956
• 负责人: Jonathan H Jaggar
• 金额: $ 49.03万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9363956
• 关键词: Arteries; Attenuated; Biotinylation; Blood Pressure; Blood Vessels; Blood flow; Bradykinin; Cardiovascular Diseases; Cell physiology; Coupling; Data; Electrophysiology (science); Endothelial Cells; Endothelium; Endothelium-Dependent Relaxing Factors; Family; Fluorescence Resonance Energy Transfer; Hypertension; Image; Immunofluorescence Immunologic; Impairment; Ion Channel; Knock-out; Knockout Mice; Ligands; Measures; Mediating; Membrane; Membrane Potentials; Methods; Modeling; Modification; Mus; Myography; Nitric Oxide; Organ; Pathologic; Physiological; Potassium; Potassium Channel; Preparation; Process; Proteins; RNA Interference; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Smooth Muscle Myocytes; Sodium; Stimulus; Systemic blood pressure; Systemic hypertension; Telemetry; Testing; Time; Vasodilation; Western Blotting; attenuation; member; normotensive; novel; patch clamp; polycystic kidney disease 1 protein; receptor

Project Summary Endothelial cells regulate arterial smooth muscle cell contractility, thereby modulating regional organ blood flow and systemic blood pressure. Dysfunctional endothelial cell control of contractility is a hallmark of several cardiovascular diseases, including hypertension, but pathological mechanisms involved are poorly understood. Membrane potential controls endothelial cell functions, but ion channels involved and functional significance are unclear. Several members of the transient receptor potential (TRP) family, are expressed in endothelial cells, but physiological functions are unclear. Similarly, several receptor ligands and stimuli are endothelium-dependent vasodilators, but their signaling mechanisms are poorly understood. Our preliminary data demonstrate that in endothelial cells, potassium (K+) channels are activated by nearby transient receptor potential (TRP) channels, leading to vasodilation. Data also suggest that K+ channel signaling is dysfunctional during hypertension. For this proposal, we will study K+ and TRP channel knockout mice. Three specific aims will be investigated. Aim 1 will test the hypothesis that K+ currents in endothelial cells regulate arterial hyperpolarization and vasodilation. Aim 2 will investigate the hypothesis that TRP channels stimulate K+ channels in endothelial cells, producing vasodilation. Aim 3 will study the hypothesis that systemic hypertension is associated with a pathological attenuation in K+ channel signaling in endothelial cells that inhibits vasodilation by these proteins. Methods used will include RT-PCR, Western blotting, biotinylation, FRET, RNAi, co-IP, immunofluorescence, patch-clamp electrophysiology, membrane potential recording, intracellular Ca2+ imaging, arterial myography and blood pressure telemetry. This proposal will provide significant novel information concerning vasoregulation by endothelial cell K+ channels.

项目摘要 内皮细胞调节动脉平滑肌细胞收缩力，从而调节区域器官血流和全身血压。功能失调的内皮细胞控制收缩力是几种心血管疾病的标志，包括高血压，但涉及的病理机制知之甚少。膜电位控制在内皮细胞功能，但涉及的离子通道和功能意义尚不清楚。瞬时受体电位（TRP）家族的几个成员在内皮细胞中表达，但生理功能尚不清楚。同样，几种受体配体和刺激是内皮依赖性的血管扩张剂，但它们的信号传导机制知之甚少。我们的初步数据表明，在内皮细胞中，钾（K+）通道被附近的瞬态受体电位（TRP）通道激活，导致血管舒张。数据还表明，在高血压期间，K+通道信号传导功能失调。 对于此建议，我们将研究K+和TRP通道敲除小鼠。将研究三个具体目标。 AIM 1将测试内皮细胞中K+电流调节动脉超极化和血管舒张的假设。 AIM 2将研究TRP通道刺激内皮细胞中K+通道的假设，从而产生血管舒张。 AIM 3将研究以下假设：系统性高血压与内皮细胞中K+通道信号中的病理衰减有关，从而抑制这些蛋白质的血管舒张。所使用的方法将包括RT-PCR，Western印迹，生物素化，FRET，RNAI，CO-IP，免疫荧光，斑块钳电生理学，膜电位记录，细胞内CA2+成像，动脉my仪和血压遥测。 该建议将提供有关内皮细胞K+通道血管调节的重要新信息。