Lipid rafts, dopamine 1 receptor, and hypertension

脂筏、多巴胺 1 受体和高血压

• 批准号: 9886774
• 负责人: Pedro A. Jose
• 金额: $ 64.24万
• 依托单位: GEORGE WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-09 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9886774
• 关键词: ADRBK1 gene; Adenylate Cyclase; Agonist; Back; Biological; Blood Pressure; C57BL/6 Mouse; Caucasians; Cell membrane; Cells; Coupled; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; DRD1 Protein; DRD1 gene; Data; Diet; Dopamine; Dopamine D1 Receptor; Dopamine Receptor; Essential Hypertension; Ethnic Origin; Ethylmaleimide; Family; G protein coupled receptor kinase; G-Protein-Coupled Receptors; Gene Frequency; Genes; Genetic Polymorphism; Golgi Apparatus; Haplotypes; Human; Hypertension; Impairment; Kidney; Knowledge; Leucine; Lipids; Maintenance; Mediating; Membrane; Membrane Microdomains; Minor; Molecular; Mus; Mutate; Mutation; NADPH Oxidase; Palmitic Acylation Site; Paraoxonase-2; Pathway interactions; Pharmacology; Phosphorylation; Phosphotransferases; Plasma; Plasma Cells; Population; Production; Protein Dephosphorylation; Protein phosphatase; Proteins; Proximal Kidney Tubules; Reactive Oxygen Species; Receptor Gene; Recycling; Regulation; Renal tubule structure; Rodent; Scaffolding Protein; Sodium Chloride; Sorting - Cell Movement; Testing; Variant; Zinc Fingers; antioxidant enzyme; blood pressure regulation; desensitization; genetic variant; high salt diet; mutant; normotensive; palmitoylation; prevent; receptor; receptor expression; receptor function; response; salt sensitive hypertension; sorting nexins; trafficking

Project Summary The D1 dopamine receptor (D1R) is important in the regulation of blood pressure (BP). Drd1 germline deletion in mice causes hypertension. G protein-coupled receptor kinase type 4 (GRK4) is important in the normal cellular recycling and function of D1R. However, some of the recycled D1R has to be targeted to lipid rafts to be functional. Thus, mutant D1Rs 347C>A and 351C>A still target to the plasma membrane but fail to localize in lipid rafts; these mutations prevent the increase in D1R-mediated stimulation of cAMP production in renal proximal tubule cells (RPTCs) and impair (e.g., 347C>A) normal BP regulation in mice. C347 and C351 in D1R are palmitoylation sites; SNX19 functions as a scaffold protein for the palmitoylation of D1R at the Golgi, via Golgi-specific zinc finger protein. Lipid rafts are also needed for the proper membrane distribution and maintenance of adenylyl cyclases 5 and 6 and regulation by D1R. Inhibition or molecular biological disruption of palmitoylation prevents D1R targeting to lipid rafts, impairs D1R function, and causes hypertension. Silencing SNX19 impairs D1R-mediated increase in cAMP and decrease in Na+ transport in RPTCs but not distal convoluted tubule cells. Renal SNX19 is important in the regulation of BP; renal-restricted silencing of Snx19 increases BP in C57Bl/6 mice on normal but not low salt diet. Germline deletion of SNX19 in mice also increases BP on normal salt diet. SNX19 is upstream of D1R. SNX19 rs2298566 is associated with decreased ability to excrete Na+ and high BP in humans and impairs D1R function in human (h)RPTCs. SNX19 protein is decreased in hRPTCs from hypertensive humans. We will test the overall hypothesis that SNX19 is important in the trafficking of D1R to lipid rafts in the RPT plasma membrane for normal D1R function. Impaired functioning of D1R in the kidney, caused by its impaired trafficking to lipid rafts in RPT plasma membranes, causes salt-sensitive hypertension. Specific Aim 1 will test the hypothesis that SNX19 targeting of D1R into the lipid rafts of RPTC plasma membrane is crucial for normal D1R function. SNX19 and GRK4 interact in lipid rafts to regulate D1R function, including D1R-mediated inhibition of Na+ transport in RPTCs. Specific aim 2 will test the hypothesis that mutations of the palmitoylation sites in the D1R gene prevent the ability of SNX19 to target the D1R to lipid rafts in the RPTC plasma membrane. Germline deletion or renal-restricted deletion of SNX19 in C57Bl/6 mice causes salt- sensitive hypertension. Mutating DRD1 to prevent D1R targeting to lipid rafts of RPTC plasma membrane also causes salt-sensitive hypertension. These genes and their proteins could be targets in the treatment of human essential hypertension.

项目摘要 D1多巴胺受体（D1R）在调节血压（BP）中很重要。 DRD1 小鼠的种系缺失会导致高血压。 G蛋白偶联受体激酶4型 （GRK4）在正常的D1R的细胞回收和功能中很重要。但是，一些 回收的D1R必须针对脂质筏具有功能。因此，突变体D1RS 347C> a和 351C>仍然是质膜的目标，但无法定位在脂质筏中；这些突变 防止肾近端小管中D1R介导的CAMP产生刺激的增加 细胞（RPTC）和损害（例如347C> a）小鼠的正常BP调节。 D1R中的C347和C351 是棕榈酰化场所； SNX19充当D1R棕榈酰化的支架蛋白 高尔基通过高尔基体特异性锌指蛋白。适当的脂质筏也需要 D1R的膜分布和维持腺苷循环酶5和6以及调节。 抑制或分子生物学破坏棕榈酰化可防止D1R靶向脂质筏， 损害D1R功能，并引起高血压。沉默SNX19损害D1R介导的 RPTC中的cAMP增加并减少了Na+转运的降低，但没有远端的曲折小管细胞。 肾脏SNX19在BP的调节中很重要。 SNX19的肾脏限制沉默增加 C57BL/6小鼠的BP在正常但低盐饮食中。小鼠SNX19的种系缺失 在正常盐饮食上增加BP。 SNX19是D1R的上游。 SNX19 RS2298566是关联的 人类排泄Na+和高bp的能力降低，并损害人类的D1R功能 （h）RPTC。高血压人的HRPTC中SNX19蛋白降低。我们将测试 总体假设SNX19在D1R到RPT中的脂质筏方面很重要 正常D1R功能的质膜。 D1R在肾脏中的功能受损，引起 由于其在RPT质膜中贩运脂质筏的贩运受损，导致盐敏感 高血压。具体目标1将检验以下假设：SNX19将D1R靶向脂质 RPTC质膜的筏对于正常的D1R功能至关重要。 SNX19和GRK4互动 在脂质筏中调节D1R功能，包括D1R介导的Na+转运的抑制 RPTC。具体目标2将检验以下假设： D1R基因阻止SNX19靶向D1R在RPTC等离子体中脂质筏的能力 膜。 C57BL/6小鼠中SNX19的种系缺失或肾脏限制缺失会导致盐 - 敏感的高血压。突变DRD1以防止D1R靶向RPTC等离子体的脂质筏 膜还会引起盐敏感的高血压。这些基因及其蛋白质可能是 治疗人类基本高血压的靶标。