PKD proteins in endothelial cells

内皮细胞中的 PKD 蛋白

基本信息

批准号：
10560613
负责人：
Jonathan H Jaggar
金额：
$ 60.23万
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：
2021-02-15 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10560613
关键词：
Acetylcholine Arteries Attenuated Biotinylation Blood Pressure Blood Vessels Blood flow Calcium Calcium Signaling Calcium-Activated Potassium Channel Cardiovascular Diseases Cell Line Cell Wall Cell physiology Coupling Data Electrophysiology (science)Endothelial Cells Endothelium Fluorescence Resonance Energy Transfer Genetic Models Hypertension Image Immunofluorescence Immunologic Impairment Knock-out Knockout Mice Ligands Light Lighting Mediating Membrane Potentials Methods Microscope Modification Mus Myography Nitric Oxide Synthase Organ PKD2 protein Pathologic Pathway interactions Physical Stimulation Physiological Process Proteins RNA Interference Receptor, Angiotensin, Type 1 Regulation Resistance Reverse Transcriptase Polymerase Chain Reaction Signal Transduction Stimulus Superoxide Dismutase Systemic blood pressure TRP channel Telemetry Tertiary Protein Structure Testing Transmembrane Domain Vasodilation Vasodilator Agents Western Blotting autocrine cell type mimetics novel patch clamp pharmacologic polycystic kidney disease 1 protein protein kinase D protein phosphatase inhibitor-2 response

项目摘要

Project Summary Endothelial cells line the wall of all blood vessels and regulate a wide variety of functions, including contractility which controls systemic blood pressure. Dysfunctional endothelial cells are a hallmark of several cardiovascular diseases, but pathological mechanisms involved are poorly understood. Endothelial cells express both PKD1 (polycystin-1), an eleven transmembrane domain protein, and PKD2 (polycystin-2), a transient receptor potential (TRP) channel. Regulatory mechanisms, physiological functions and pathological involvement during hypertension of PKD1 protein and PKD2 channels in endothelial cells are unclear. Using a wide variety of approaches and inducible, endothelial cell-specific knockout mice, we provide evidence that physical coupling of PKD1 to PKD2 in endothelial cells stimulates vasodilation. Preliminary data also suggest that PKD1/PKD2 channel signaling is dysfunctional during hypertension, which attenuates this vasodilatory signaling mechanism. In this proposal, we will investigate three specific aims. Aim 1 will test the hypothesis that endothelial-dependent physiological stimuli activate PKD1/PKD2 coupling in endothelial cells, leading to vasodilation. Aim 2 will investigate the mechanisms by which endothelial-dependent stimuli activate PKD1/PKD2 channels in endothelial cells to produce vasodilation. Aim 3 will study the hypothesis that hypertension is associated with pathological alterations in PKD1/PKD2 channel signaling in endothelial cells that inhibits vasodilation mediated 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 project will provide significant novel information concerning vasoregulation by endothelial cell PKD1 and PKD2 proteins.

项目摘要内皮细胞在所有血管的壁上排列并调节各种功能，包括收缩力控制系统性血压。功能失调的内皮细胞是几个的标志心血管疾病，但涉及的病理机制知之甚少。内皮细胞表达PKD1（Polycystin-1），十一跨膜结构蛋白和PKD2（Polycystin-2），A 瞬态受体电位（TRP）通道。调节机制，生理功能和病理在内皮细胞中PKD1蛋白和PKD2通道高血压期间的参与尚不清楚。使用各种各样的方法和可诱导的内皮细胞特异性敲除小鼠，我们提供了证据表明 PKD1与内皮细胞中PKD2的物理耦合刺激血管舒张。初步数据也暗示在高血压期间，PKD1/PKD2通道信号传导功能失调，这会衰减这种血管舒张信号传导机制。在此提案中，我们将研究三个具体目标。 AIM 1将检验假设内皮依赖性的生理刺激激活了内皮细胞中的PKD1/PKD2耦合，导致血管舒张。 AIM 2将研究内皮依赖性刺激激活的机制内皮细胞中的PKD1/PKD2通道产生血管舒张。 AIM 3将研究以下假设高血压与内皮细胞中PKD1/PKD2通道信号传导的病理改变有关这抑制了这些蛋白质介导的血管舒张。所使用的方法将包括RT-PCR，Western blotting，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，动脉典范和血压遥测。这个项目将提供有关内皮细胞PKD1和PKD2蛋白质血管调节的重要新信息。