Vascular Cell Regulation of Connexins

描述（由申请人提供）：内皮细胞（EC）和平滑肌细胞（SMC）之间的细胞-细胞通信，由间隙连接提供，协调血管收缩性和电传导。血管间隙连接蛋白（connexins (Cx)40和Cx43）的基因敲除（KO）会产生显著不同的血压和传导。这些结果的一个可能的原因是1)Cx40和Cx43的共同调节，这实际上会2)改变通过血管壁间隙连接的细胞-细胞通信的方法。基于这一想法，我假设EC和SMC之间的接触在调节连接蛋白的功能和表达，从而调节血管反应性方面起着关键作用。为了阐明这一假设，将开发和利用一个类似于体内的体外系统来模拟EC和SMC的相互作用。该模型将从同一小鼠和主动脉切片中分离EC和SMC细胞，用于测试EC和SMC之间的染料和第二信使传递方法。这些结果将与原位野生型和连接蛋白KO动物的类似实验进行比较。结合这些方法，将有助于描述EC和SMC之间连接蛋白的生理作用。

DESCRIPTION (provided by applicant): Cell-cell communication between endothelial cells (EC) and smooth muscle cells (SMC), provided by gap junctions, coordinates vessel contractility and electrical conduction. Gene knockout (KO) of vessel gap junction proteins (connexins (Cx)40 and Cx43) yields dramatically different blood pressures and conduction. A possible reason for these results are 1) co-regulation of Cx40 and Cx43, which in effect would 2) alter the method of cell-cell communication by gap junctions in the vessel wall. Based on this idea, I hypothesize that contact between EC and SMC plays a key role in modulating connexin function and expression, and thus in modulating vascular reactivity. In order to elucidate this hypothesis, an in vitro system to model EC and SMC interactions similar to in vivo will be developed and utilized. This model, in which EC and SMC cells will be isolated from the same mouse and aortic sections, will be used to test methods of dye and second messenger transfer between EC and SMC. These results will be compared to similar experiments on in situ wild type and connexin KO animals. Combined, these methods will help delineate the physiological role of connexins between EC and SMC.