PROTEASE-ACTIVATED RECEPTORS IN EMBRYONIC DEVELOPMENT

胚胎发育中的蛋白酶激活受体

• 批准号: 6642830
• 负责人: SHAUN R. COUGHLIN
• 金额: $ 36.88万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6642830
• 关键词: G protein coupled receptor kinase; biological signal transduction; blood coagulation; blood vessels; cell type; developmental genetics; electron microscopy; embryo /fetus tissue /cell culture; embryogenesis; fibrinogen; gene targeting; genetic enhancer element; genetic promoter element; genetically modified animals; histochemistry /cytochemistry; histogenesis; in situ hybridization; laboratory mouse; messenger RNA; protein structure function; receptor expression; serine proteinases; thrombin receptor; thrombomodulin; vascular endothelium; G protein coupled receptor kinase; biological signal transduction; blood coagulation; blood vessels; cell type; developmental genetics; electron microscopy; embryo /fetus tissue /cell culture; embryogenesis; fibrinogen; gene targeting; genetic enhancer element; genetic promoter element; genetically modified animals; histochemistry /cytochemistry; histogenesis; in situ hybridization; laboratory mouse; messenger RNA; protein structure function; receptor expression; serine proteinases; thrombin receptor; thrombomodulin; vascular endothelium

DESCRIPTION: (Adopted from the Applicant's abstract.) The long-term goal of this research is to understand the role of the coagulation system in development. Considerable evidence exists from knock-outs of components of that system that it has an important role in embryogenesis, including evidence from PAR-1 knock-out, which leads to the demise of approximately half of PAR1-deficient embryos at aboutE9.5. Death of these embryos was associated with bleeding but could not be attributed to defects in platelet function, implying that thrombin signaling may have a role in embryonic development unrelated to its role in hemostasis, at least in the usual sense. In situ hybridization revealed PAR1 mRNA expression in endocardial and endothelial cells at E9.5, suggesting that PAR1 signaling might be important for normal blood vessel development and vascular integrity. The Principal Investigator postulates that the coagulation cascade functions in part as a "leak detector" that monitors and regulates the integrity of developing blood vessels. To test these hypotheses he proposes the following specific aims. Aim 1 will ask the question: In which cell type(s) is PAR1 signaling important for embryonic development? To address this question they will: a) Use a lacZ knock-in to identify the cell types that express PAR1 and determine their fate in PAR1-deficient embryos; b) Characterize vascular development in PAR1-deficient embryos; c) determine by transgenic rescue whether lack of PAR1 function in endothelial cells is the primary defect in PAR1-deficient embryos; d) Use knock-in of gain-of-function mutations to identify the cells in which PAR1 activation normally occurs during embryonic development; e) Test the hypothesis that disinhibited thrombin production and PAR1 signaling are responsible for the death of thrombomodulin-deficient embryos. In Aim 2, they ask the question: Beyond PAR1, what are the other effectors of the coagulation cascade during embryonic development? They will: a) Test the hypothesis that fibrinogen becomes important in the absence of PAR1, b) Determine whether PAR4 and/or as yet unidentified receptors provide "back-up" thrombin signaling in embryonic development, c) Determine the effect of PAR2 deficiency in combination with other PAR-deficiencies on embryonic development.

描述：（从申请人的摘要中采用。） 这项研究是了解凝血系统在 发展。大量的证据来自该组成部分的敲除 它在胚胎发生中具有重要作用的系统，包括 PAR-1淘汰赛，导致大约一半的灭亡 PAR1缺陷胚胎约9.5。这些胚胎的死亡与 出血，但不能归因于血小板功能的缺陷，这意味着 该凝血酶信号传导可能在与 它在止血中的作用至少在通常的意义上。原位杂交 揭示了在E9.5处的内膜和内皮细胞中的PAR1 mRNA表达， 表明PAR1信号对于正常血管可能很重要 发展和血管完整性。主要调查员假定 凝血级联反应部分作为监测的“泄漏检测器” 并调节发育血管的完整性。测试这些 他提出了以下特定目标。 AIM 1会问 问题：在哪种细胞类型中，PAR1信号对于胚胎很重要 发展？为了解决这个问题，他们将：a）使用lacz敲击 识别表达PAR1的细胞类型并确定其命运 PAR1缺陷胚胎； b）表征PAR1缺陷的血管发育 胚胎； c）通过转基因拯救确定缺乏PAR1功能是否功能 内皮细胞是PAR1缺陷胚胎中的主要缺陷。 d）使用 敲入功能收益突变，以识别PAR1的细胞 激活通常在胚胎发育过程中发生。 e）检验假设 抑制的凝血酶产生和PAR1信号是负责的 血小板结构蛋白缺陷的胚胎死亡。在AIM 2中，他们提出了一个问题： 除了PAR1之外，凝血级联的其他效应子是什么 胚胎发展？他们将：a）检验纤维蛋白原的假设 在没有par1的情况下变得很重要，b）确定PAR4和/或AS是否 然而，身份不明的受体在胚胎中提供“备份”凝血酶信号传导 开发，c）确定PAR2缺乏症与结合的影响 其他有关胚胎发展的缺陷。