Physiological regulation of vascular function by mutant caveolin-1

描述(由申请人提供)：内源性小窝蛋白-1(CAV-1)对eNOS功能的抑制在体外和体内都有很好的特征。我们进一步剖析了eNOS和Cav-1之间的相互作用，并通过突变小窝蛋白支架结构域(CSD)解离了Cav-1与eNOS的抑制结合。AP-Cav 3 PM是一种细胞通透性多肽，含有Cav-1 CSD的突变形式，在体外和体外都能激活eNOS。重要的是，我们现在已经开发出可诱导的内皮细胞特异性Cav-1转基因小鼠，它具有Cav-1(F92ACav-1)的突变形式，通过Cav-1和eNOS的解离促进NO的释放。考虑到这一点，了解eNOS和Cav-1之间的相互作用将有助于从分子上剖析Cav-1作为eNOS的负调控因子的作用以及它在血管生成中的作用。我们假设F92ACav-1突变小鼠除了NO产生增加、血压降低、血管反应性降低和血管生成潜力增加外，还将表现出结构正常的小窝和Cav-1分布。令人兴奋的是，这一提议提供了第一个体内平台，用于研究Cav-1作为小窝的主要结构成分的生理作用与其作为信号平台的作用。为了更详细地研究这一重要相互作用的调节，我们将确定：1.F92ACav-1突变对体内心血管表型的影响；2.F92ACav-1突变对体外血管系统生理反应的影响；3.F92ACav-1突变如何影响体内血管生成。

DESCRIPTION (provided by applicant): Inhibition of eNOS function by endogenous caveolin-1 (Cav-1) has been well characterized in vitro and in vivo. We have further dissected the interaction between eNOS and Cav-1, and have dissociated the inhibitory binding of Cav-1 to eNOS using mutagenesis of the caveolin scaffolding domain (CSD). AP-Cav 3PM, a cell permeable peptide harboring a mutant form of the Cav-1 CSD, activates eNOS in vitro and ex vivo. Importantly we have now developed inducible, endothelial cell specific Cav-1 transgenic mice that have a mutant form of Cav-1 (F92A Cav-1) that promotes NO release through the dissociation of Cav-1 and eNOS. With this in mind, understanding the interactions between eNOS and Cav-1 will permit molecular dissection of the roles of Cav-1 as a negative regulator of eNOS and the role this plays angiogenesis. We hypothesize that F92A Cav-1 mutant mice will exhibit structurally normal caveolae and Cav-1 distribution, in addition to increased NO production, reduced blood pressure, lower vascular reactivity, and increased angiogenic potential. Excitingly, this proposal presents the first in vivo platform for the study of Cav-1 physiological role as the major structural component of caveolae versus its role as a signaling platform. To examine the regulation of this important interaction in more detail, we will determine:1 The effects of the F92A Cav-1 mutation on the cardiovascular phenotype in vivo; 2. The effects of the F92A Cav-1 mutation on physiologic responses of the vasculature ex vivo and 3. How the F92A Cav-1 mutation affects angiogenesis in vivo.