PKD proteins in endothelial cells

内皮细胞中的 PKD 蛋白

• 批准号: 10339327
• 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
• 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/10339327
• 关键词: Acetylcholine; Arteries; Attenuated; Biotinylation; Blood Pressure; Blood Vessels; Blood flow; Calcium; 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; Pharmacology; Physiological; Potassium Channel; 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; Wnt proteins; autocrine; cell type; mimetics; novel; patch clamp; polycystic kidney disease 1 protein; protein kinase D; protein phosphatase inhibitor-2; response

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 内皮细胞排列在所有血管的壁上，调节各种功能，包括收缩性 控制全身血压功能障碍的内皮细胞是几种 心血管疾病，但涉及的病理机制知之甚少。内皮细胞 表达PKD 1（多囊蛋白-1），一种十一跨膜结构域蛋白，和PKD 2（多囊蛋白-2），一种 瞬时受体电位（TRP）通道。调节机制、生理功能和病理 高血压期间内皮细胞中PKD 1蛋白和PKD 2通道的参与尚不清楚。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，我们提供的证据， 内皮细胞中PKD 1与PKD 2的物理偶联刺激血管舒张。初步数据还表明， PKD 1/PKD 2通道信号在高血压期间功能失调，这减弱了这种血管舒张作用， 信号机制在本建议中，我们将探讨三个具体目标。目标1将检验假设 内皮依赖性生理刺激激活内皮细胞中的PKD 1/PKD 2偶联，导致 血管舒张目的2将探讨内皮依赖性刺激激活 PKD 1/PKD 2通道在内皮细胞中产生血管舒张。目标3将研究假设， 高血压与内皮细胞PKD 1/PKD 2通道信号转导的病理改变相关 抑制由这些蛋白质介导的血管舒张。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，膜片钳电生理，膜电位 记录、细胞内Ca 2+成像、动脉肌电图和血压遥测。该项目将 提供了关于内皮细胞PKD 1和PKD 2蛋白的血管调节的重要新信息。