Decoding gap junction communication under shear stress

描述(申请人提供)：在内皮中，人们对通过缝隙连接调节细胞信号以响应血流动力学剪切力的机制知之甚少。相对较少的研究研究了单个缝隙连接蛋白在流体诱导的流体剪应力作用下的表达和功能。尽管一氧化氮(NO)已被证明在静态细胞培养中调节缝隙连接通讯，但剪切力诱导的NO对这种通讯的调节尚未被探索。需要研究的问题是，在不同的血流动力学机制(层流或紊乱)中，这种类金刚蛋白的释放如何调节不同的连接蛋白。此外，在没有细胞骨架的情况下，流体剪应力如何影响细胞间通讯的问题也需要研究，因为细胞骨架蛋白被推测参与缝隙连接组装。这项建议中概述的策略将使我们能够利用新开发的技术，如siRNA和长寡核苷酸微阵列，阐明血管壁重塑过程中内皮细胞缝隙连接的生理调节机制，以响应各种血流动力学剪切力。

DESCRIPTION (provided by applicant): In the endothelium very little is known about the mechanisms that are involved in regulating cellular signaling via gap junctions in response to hemodynamic shearing forces. Relatively few studies have investigated the expression and function of individual gap junction proteins in response to flow-induced fluid shear stress. Although nitric oxide (NO) has been shown to modulate the gap junction communication in static cell culture, the regulation of shear induced NO on this communication has not yet been explored. The question of how this autacoid release regulates different connexins in the various hemodynamic schemes (laminar or disturbed) needs to be investigated. In addition, the question of how fluid shear stress affects intercellular communication in the absence of cytoskeleton also needs to be studied, since cytoskeletal proteins are speculated to be involved in gap junction assembly. The strategy outlined in this proposal will allow us to elucidate the poorly understood physiological regulatory mechanisms of gap junctions in the endothelium during vessel wall remodeling in response to various hemodynamic shearing forces using newly developed techniques such as siRNA and Long Oligo microarrays.