Adrenergic and Purinergic Regulation of Target Cells

靶细胞的肾上腺素能和嘌呤能调节

• 批准号: 7730124
• 负责人: PAUL A INSEL
• 金额: $ 30.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-20 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7730124
• 关键词: ATP Receptors; Adrenal Glands; Adrenergic Agents; Adrenergic Receptor; Canis familiaris; Cardiovascular system; Catecholamines; Caveolae; Caveolins; Cell surface; Cells; Chromaffin Cells; Data; Disease; Duct (organ) structure; Ductal Epithelium; Engineering; Environment; Epinephrine; Epithelial; Epithelial Cells; Epithelium; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-substrate; GTP-Binding Proteins; Goals; Heterotrimeric GTP-Binding Proteins; Hypertension; Ion Channel; Kidney; Knock-out; Knockout Mice; Link; Liquid substance; MDCK cell; Mediating; Membrane Microdomains; Modification; Molecular; Multiple Abnormalities; Mus; Nerve; Norepinephrine; Nucleotides; P2X-receptor; P2Y2 receptor; Pharmaceutical Preparations; Purinoceptor; Regulation; Renal function; Resistance; Role; Signal Pathway; Signal Transduction; Signaling Protein; Sodium Chloride; Sympathetic Nervous System; System; Testing; Tubular formation; Water; adrenergic; base; caveolin-2; cell growth regulation; cell injury; collecting tubule structure; design; extracellular; genetically modified cells; human CCR10 protein; insight; novel; public health relevance; receptor; research study; response; scaffold; tissue/cell culture

DESCRIPTION (provided by applicant): This renewal application seeks to continue studies that use Madin Darby Canine Kidney (MDCK) cells and knockout mice to assess molecular and functional aspects of adrenergic and P2Y (nucleotide, purinergic) receptors. The proposed studies will test hypotheses related to findings that we have obtained in MDCK cells and more recently in mouse primary kidney collecting duct (mpkCCD) cells as well as in P2Y2 knockout mice, which we have recently shown have salt-resistant hypertension and multiple abnormalities in renal function. The Specific Aims involve studies of: 1) Expression and compartmentation in raft/caveolar microdomains of G protein-coupled receptor/G-proteins/effectors of adrenergic and P2Y receptors in MDCK and mpkCCD cells and assessment of sumoylation (a novel modification that we have recently identified) of caveolins; 2) Functional roles of renal P2Y and adrenergic receptors using receptor-knockout mice. The combined use of cultured renal epithelial cells and knockout mice provides a complementary strategy that is designed to reveal new insights regarding the regulation of epithelial cells and renal function by adrenergic and P2Y receptors. The findings may have implications for cardiovascular, renal and other disorders that involve the sympathetic nervous system or in which cell injury leads to release of nucleotides. PUBLIC HEALTH RELEVANCE: This project will assess 2 important classes of proteins, receptors for adrenaline (and adrenaline-like drugs) and for a major cell constituent, ATP. The receptors, which are located on the surface of cells, recognize those molecules in the extracellular environment but then alter intracellular function. Experiments are proposed in: 1) tissue culture cells to assess the organization of the components involved in information transfer across the cell surface and 2) mice engineered to have loss in receptors for ATP or adrenaline with the goal of understanding how the receptors regulate fluid and salt handling, in particular by the kidney.

描述（由申请人提供）：本更新申请旨在继续使用 Madin Darby 犬肾 (MDCK) 细胞和基因敲除小鼠来评估肾上腺素能和 P2Y（核苷酸、嘌呤能）受体的分子和功能方面的研究。拟议的研究将测试与我们在 MDCK 细胞、最近在小鼠原代肾集合管 (mpkCCD) 细胞以及 P2Y2 敲除小鼠中获得的发现相关的假设，我们最近显示这些小鼠具有耐盐性高血压和多种异常在肾功能方面。具体目标涉及以下研究： 1) MDCK 和 mpkCCD 细胞中 G 蛋白偶联受体/G 蛋白/肾上腺素能和 P2Y 受体的筏/穴微结构域的表达和区隔以及 sumoylation 的评估（我们的一种新修饰）最近发现的）小窝蛋白； 2) 使用受体敲除小鼠研究肾 P2Y 和肾上腺素能受体的功能作用。培养的肾上皮细胞和基因敲除小鼠的联合使用提供了一种补充策略，旨在揭示有关肾上腺素能受体和 P2Y 受体调节上皮细胞和肾功能的新见解。这些发现可能对心血管、肾脏和其他涉及交感神经系统或细胞损伤导致核苷酸释放的疾病有影响。 公共健康相关性：该项目将评估两类重要的蛋白质：肾上腺素（和肾上腺素样药物）受体和主要细胞成分 ATP 受体。位于细胞表面的受体识别细胞外环境中的这些分子，然后改变细胞内功能。建议进行以下实验：1) 组织培养细胞，以评估参与跨细胞表面信息传递的成分的组织；2) 小鼠经过基因改造，使其 ATP 或肾上腺素受体丧失，目的是了解受体如何调节体液和肾上腺素。盐的处理，特别是通过肾脏。