Isakson Brant E, Duling Brian R
Cardiovascular Research Center, University of Virginia School of Medicine, Charlottesville 22908, USA.
Circ Res. 2005 Jul 8;97(1):44-51. doi: 10.1161/01.RES.0000173461.36221.2e. Epub 2005 Jun 16.
Heterocellular communication between vascular smooth muscle cells (VSMC) and endothelial cells (EC) at the myoendothelial junction (MEJ) is a critical part of control of the arteriolar wall. We have developed an in vitro model of the MEJ composed of primary cultures of murine EC and VSMC. Immunoctytochemistry and immunoblots demonstrated Cx37 and Cx43 in both cell types, whereas Cx40 was found only in EC. Cx37 was excluded from the MEJ in both EC and VSMC. Connexin composition as well as functionality of the gap junctions at the MEJ was assessed by measuring diffusional transfer of biocytin and Cy3. Using connexin-specific blockers and manipulations of expression of individual connexin proteins, we confirmed that Cx37 is not a part of EC-VSMC coupling, and we demonstrated that heterotypic gap junctions are functional at the MEJ. We speculate that specific gap junction organization may be a vital component of EC-VSMC contact at the MEJ.
肌内皮连接(MEJ)处血管平滑肌细胞(VSMC)与内皮细胞(EC)之间的异细胞通讯是小动脉壁控制的关键部分。我们构建了一个由小鼠EC和VSMC原代培养物组成的MEJ体外模型。免疫细胞化学和免疫印迹显示两种细胞类型中均存在Cx37和Cx43,而Cx40仅在EC中发现。在EC和VSMC中,Cx37均被排除在MEJ之外。通过测量生物素和Cy3的扩散转移来评估MEJ处缝隙连接的连接蛋白组成及其功能。使用连接蛋白特异性阻滞剂和对单个连接蛋白表达的操作,我们证实Cx37不是EC-VSMC偶联的一部分,并且我们证明异型缝隙连接在MEJ处具有功能。我们推测特定的缝隙连接组织可能是MEJ处EC-VSMC接触的重要组成部分。