权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

SK3 channel trafficking in endothelial cells

内皮细胞中的 SK3 通道运输

基本信息

批准号：
10275918
负责人：
Jonathan H Jaggar
金额：
$ 63.6万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10275918
关键词：
Arteries Attenuated Biotinylation Blood Blood Pressure Blood Vessels Calcium Cardiovascular Diseases Cell Line Cell membrane Cells Data Electrophysiology (science)Endothelial Cells Fluorescence Resonance Energy Transfer Functional disorder Genes Genetic Human Hypertension Image Immunofluorescence Immunologic Ion Channel Knockout Mice Mediating Membrane Potentials Membrane Proteins Methods Microscopy Modeling Mus Mutation Myography Pathologic Pathway interactions Patients Physiological Potassium Probability Process Proteins RNA Interference Resolution Signal Transduction Stimulus Surface Systemic blood pressure Systemic hypertension TRP channel Telemetry Testing Transgenic Mice Urine Vanilloid Vasodilation Vasodilator Agents Western Blotting familial hypertension fluvastatin inhibitor/antagonist mouse model mutant novel patch clamp rab GTP-Binding Proteins receptor stem trafficking vasoconstriction

项目摘要

Project Summary Endothelial cells line the wall of all blood vessels and regulate a wide variety of functions, including contractility and systemic blood pressure. Endothelial cell dysfunction is a hallmark of several cardiovascular diseases, but pathological mechanisms involved are unclear. Endothelial cells express small-conductance calcium-activated potassium (SK3) channels, which regulate contractility. Currents (I) generated by SK3 channels in endothelial cells is the product of the number of channels (N), their open probability (PO) and amplitude (i), such that I=N.PO.i. Previous studies have focused on identifying mechanisms that regulate the activity of surface SK channels in endothelial cells. In contrast, mechanisms that control the number (N) of surface SK3 channels in endothelial cells are poorly understood. Importantly, it is unclear whether physiological stimuli regulate the abundance of surface SK3 channels to alter arterial contractility. Similarly uncertain is if pathological alterations in the control of surface SK3 channel abundance occurs during hypertension, leading to attenuated vasodilation. Using a wide variety of approaches, we provide evidence that vasodilator stimuli activate trafficking mechanisms that rapidly increase surface SK3 channel abundance in endothelial cells to induce vasodilation. Preliminary data also suggest that SK3 channel trafficking is dysfunctional during hypertension, which attenuates this vasodilatory signaling mechanism. In this proposal, we will investigate three specific aims. Aim 1 will investigate signaling mechanisms by which physiological stimuli alter the surface abundance of SK3 channels and examine their functional significance. Aim 2 will examine trafficking mechanisms that control surface SK3 channels in endothelial cells to modify arterial contractility. Aim 3 will study the hypothesis that hypertension is associated with pathological alterations in SK3 channel trafficking in endothelial cells that inhibit vasodilation by these proteins. Methods used will include biotinylation, Western blotting, FRET, RNAi, co-IP, immunofluorescence, super-resolution microscopy, patch-clamp electrophysiology, membrane potential recording, intracellular Ca2+ imaging, arterial myography, blood pressure telemetry and transgenic mice. This project will provide significant novel information concerning vasoregulation by SK3 channel trafficking in endothelial cells.

项目总结