Functional Analysis of the Prostaglandin EP3 Receptor

前列腺素 EP3 受体的功能分析

• 批准号: 6624506
• 负责人: RICHARD M. BREYER
• 金额: $ 28.88万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6624506
• 关键词: RNA splicing; biological signal transduction; cyclic AMP; eicosanoid metabolism; enzyme linked immunosorbent assay; genetic mapping; genetic transcription; laboratory mouse; messenger RNA; molecular cloning; phosphorylation; prostaglandin E; prostaglandin receptor; protein isoforms; protein structure function; receptor coupling; receptor expression; renal tubular transport; tissue /cell culture; transfection; video microscopy; western blottings

DESCRIPTION (provided by applicant): Prostaglandin E2 (PGE2) is a major cyclooxygenase metabolite of arachidonic acid and a potent modulator of a wide variety of cellular responses including vascular tone, febrile response as well as water and ion transport in the kidney. There is now firm evidence that most of PGE2's effects are mediated via specific guanine nucleotide regulatory protein coupled receptors designated EP1, EP2, EP3, and EP4. The EP3 receptor is the unique among the EP receptor family in that it is represented by multiple alternatively spliced variants generated by alternative splicing from a single gene of a common precursor mRNA. The principal hypothesis of this proposal is that differential expression of EP3 receptor splice variants on individual cell types results in differential activation of signaling pathways that determine the physiologic response of a target cell to PGE2. Implicit in this hypothesis is the notion that the EP3 receptor variants have functionally distinct tissue distribution, cellular expression, and intracellular compartmentalization as well as distinct signal transduction properties. To test this hypothesis, in Specific Aim 1 we will determine the physiologic role of the EP3 receptor splice variants using an genetically modified mice strategy expressing a restricted repertoire of EP3 receptor splice variants. We will characterize phenotypic changes focusing on blood pressure, and vascular reactivity. Although the EP3 receptor has classically been characterized as a Gi coupled receptor that signals by lowering intracellular cAMP levels, recent studies suggest that EP3 alternative splice variants couple through other non-Gi signal transduction pathways. In Specific Aim 2, studies will focus on the characterization of the EP3 receptor-mediated mechanism of cAMP independent signal transduction and its effect on gene transcription. We will examine the potential for this pathway to modulate gene expression using specific inhibitors and phosphorylation-specific antibodies in Western blot analysis, and promoter analysis.

描述（申请人提供）：前列腺素E2（PGE2）是主要的 花生四烯酸的环氧合酶代谢物和宽的有效调节剂 各种细胞反应，包括血管张力，发热反应 作为肾脏中的水和离子运输。现在有大多数证据表明 PGE2的作用是通过特异性鸟嘌呤核苷酸调节介导的 蛋白质偶联受体指定EP1，EP2，EP3和EP4。 EP3受体 是EP受体家族中的独特之处，因为它由 通过替代拼接生成的多个剪接变体 一个常见前体mRNA的单个基因。主要假设 提案是EP3受体剪接变体的差异表达在 单个细胞类型导致信号通路的差异激活 这决定了靶细胞对PGE2的生理反应。隐含 该假设是EP3受体变体在功能上的观念 独特的组织分布，细胞表达和细胞内 隔室化以及不同的信号转导特性。到 检验该假设，在特定目的1中我们将确定生理作用 使用基因修饰的小鼠策略的EP3受体剪接变体的 表达EP3受体剪接变体的受限曲目。我们将 表征表型变化的特征，重点是血压和血管 反应性。尽管EP3受体在经典上被描述为 GI偶联受体通过降低细胞内营地的信号，最近 研究表明，EP3替代剪接变体通过其他 非GI信号转导途径。在特定目标2中，研究将重点放在 CAMP独立的EP3受体介导的机制的表征 信号转导及其对基因转录的影响。我们将检查 使用特定的这种途径调节基因表达的潜力 蛋白质印迹分析中的抑制剂和磷酸化特异性抗体， 和启动子分析。