Molecular Genetics of the ET-1/AngII Receptor

ET-1/AngII 受体的分子遗传学

• 批准号: 6852637
• 负责人: NELSON RUIZ-OPAZO
• 金额: $ 40.38万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6852637
• 关键词: angiotensin /renin /aldosterone hypertension; angiotensin II; angiotensin receptor; endothelin; gender difference; gene targeting; genetic susceptibility; genetically modified animals; hypertension; laboratory mouse; laboratory rat; protein isoforms; receptor expression

DESCRIPTION (provided by the applicant): Endothelin-1 (ET-1) and angiotensin II (AngII) are critical hormones necessary for the appropriate function of the cardiovascular system, acting centrally as well as peripherally. Both hormonal systems have been implicated in cardiovascular pathology -- hypertension, endothelial dysfunction, cardiac hypertrophy and failure. Several isoreceptors for both ET-1 and AngII have been recently characterized. Of these, the dual ET-1/AngII receptor, isolated by us via an approach based on and supporting the molecular recognition theory elucidates novel information providing the bases for new hypotheses and questions. The long-term objective of this proposal is to dissect the specific contribution of the ET-l/AngII receptor gene to blood pressure regulation in normal and pathophysiological states like hypertension. In this research proposal, we will test the following hypotheses: 1) Based on our preliminary data showing a) gender-bias o 1I the cosegregation of the ET-II /AngII receptor locus with hypertension in an F2 (Dahl S x Dahl R) intercross, and b) structural differences between the Dahl salt-sensitive and the Dahl salt-resistant rat ET-1/AngII receptors, we hypothesize that the ET-1/AngII receptor acts in a gender specific manner influencing blood pressure in the Dahl rat model. 2) Based on the detected abundance of the ET-l/AngII receptor mRNA in brain and cardiovascular tissues, we hypothesize that this receptor is a prominent physiologic target for ET- 1 and AngII in these organs, and therefore plays an important role in blood pressure regulation. Investigating the following specific aims will test these hypotheses: 1. Gender specific role of the ET-1/AngII receptor in susceptibility to salt-sensitive hypertension. 1A) Functional characterization of the Dahl S and Dahl R ET-1/AngII receptors that differ in two amino acids. 1B) Establishment of three inbred trans genie Dahl S rat lines bearing exogenous Dahl R ET-1/AngII receptor, Tg[RET-1/AngIIr], and three control Dahl S rat lines bearing exogenous Dahl S ET- 1/AngIl receptor, Tg[SET- 1/AnglIr], driven by the cognate Dahl R 5' flanking regulatory region. 1C) Determine the transgenic molecular phenotype, that is the relative amounts of exogenous (transgenic) Dahl R and Dahl S ET- 1/Angll receptor mRNA and endogenous Dahl S ET-1/AngIl receptor mRNA expression levels in Tg[RET-1/AngIIr] and Tg[SET-1/AngIIr} rats respectively. 1D) Determine the physiological effects of the Dahl R and Dahl S ET- 1/AngII receptor transgene by analyzing differences between age-matched non-transgenic inbred Dahl S rats (males and females) and transgenic inbred Tg[RET- 1/AnglIr] and Tg[SET1I AngIIr] Dahl S rat lines with respect to 1) the onset, course and degree of hypertension; 2) lifespan; 3) severity of hypertensive renal disease. 2. Dissection of physiologic role(s) of the ET-1/AngII receptor. 2A) Tissue specific, spatial and temporal expression patterns in development of the ET-1/AngII receptor gene. 2B) Targeted disruption of the ET- 1/AngIl receptor gene and development of "knockout" mouse model in order to define its physiologic role in an integrated biologic experimental system. Our proposed studies will elucidate the physiological role of the ET- 1 /AngII receptor throughout development. Furthermore, it will demonstrate the gender specific involvement of the ET- 1/AngII receptor in salt-sensitive hypertension in the Dahl rat model, thus providing a validated prioritization scheme to investigate the potential role of this receptor in human essential hypertension.

描述（申请人提供）：内皮素-1（ET-1）和血管紧张素II （Angii）是适当功能的关键激素 心血管系统，集中作用以及外围。两者都是激素 系统与心血管病理学有关 - 高血压， 内皮功能障碍，心脏肥大和衰竭。几个异感受器 对于ET-1和ANGII，最近都被表征了。其中，双重 ET-1/ANGII受体，由我们通过基于和支持的方法隔离 分子识别理论阐明了提供基础的新信息 用于新的假设和问题。该提议的长期目标是 剖析ET-L/ANGII受体基因对血液的特定贡献 正常和病理生理状态（如高血压）的压力调节。 在这项研究建议中，我们将测试以下假设：1）基于 我们的初步数据显示a）性别偏见o 1i ET-II的cosegregation /AngII受体基因座，在F2（Dahl S X Dahl R）中有高血压， b）dahl盐敏感与dahl之间的结构差异 耐盐的大鼠ET-1/ANGII受体，我们假设ET-1/ANGII 受体以性别特定方式作用，影响血压 达尔大鼠模型。 2）基于检测到的ET-L/ANGII受体的丰度 大脑和心血管组织中的mRNA，我们假设该受体是 这些器官中的ET 1和Angii的突出生理靶标，以及 因此，在血压调节中起着重要作用。调查 以下具体目的将检验以下假设：1。特定角色 ET-1/ANGII受体的易感性敏感性高血压。 1A）DAHL S和DAHL R ET-1/ANGII受体的功能表征 这在两个氨基酸中有所不同。 1b）建立三个近交式精灵 DAHL S带有外源性DAHL R ET-1/ANGII受体的大鼠系 Tg [ret-1/angiir]和三个控制Dahl S带有外源性Dahl s的大鼠线 ET-1/Angil受体，TG [Set-1/Anglir]，由同源dahl r 5'驱动 侧翼监管区域。 1C）确定转基因分子表型， 这就是外源（转基因）dahl r和dahl s et-的相对量 1/Angll受体mRNA和内源性DAHL S ET-1/Angil受体mRNA表达 TG [RET-1/Angiir]和Tg [set-1/angiir}大鼠的水平分别。 1D） 确定dahl r和dahl s et-1/angii的生理效应 受体转基因通过分析年龄匹配的非转基因之间的差异 近交dahl s大鼠（男性和女性）和转基因的近百亲tg [ret-1/anglir] 和Tg [set1i angiir] Dahl s鼠线相对于1）发作，过程和 高血压程度； 2）寿命； 3）高血压肾脏疾病的严重程度。 2。ET-1/ANGII受体的生理作用的解剖。 2a）组织 特定的，空间和时间表达模式的发展 ET-1/ANGII受体基因。 2b）靶向破坏ET-1/Angil受体 基因和“敲除”小鼠模型的开发，以定义其 在综合生物实验系统中的生理作用。我们提出的 研究将阐明ET-1 /Angii受体的生理作用 整个发展。此外，它将展示特定的性别 ET-1/ANGII受体参与盐敏感高血压 DAHL大鼠模型，因此提供了经过验证的优先级方案 该受体在人类基本高血压中的潜在作用。