FUNCTION OF CX43-INTERACTING PROTEINS IN NEURAL CREST MIGRATION

适当协调神经脊迁移是正常心血管的绝对要求 发育--尤其是流出道的形态发生。缝隙连接蛋白Cx43有一个关键 在心脏神经脊细胞的定向迁移中起作用。Cx43的分子机制 影响神经脊运动尚不清楚。推动细胞迁移的力量主要是由 肌动蛋白细胞骨架的动力学。我们假设，介导连接蛋白和连接蛋白之间联系的蛋白质 肌动蛋白是调节细胞运动的候选蛋白。一种已知的调节相互作用的分子 缝隙连接蛋白和肌动蛋白是ZO1--一种与果蝇相关的PDZ蛋白，大的抑癌蛋白圆盘。 古尔迪实验室已经证明，Cx43-ZO1的相互作用对于发育重建的程度至关重要 心肌细胞间的缝隙连接接触。此外，我们有数据表明，ZO1的抑制- Cx43相互作用降低成纤维细胞和上皮细胞的运动性 在“划痕”迁移实验中培养的单层细胞。我们将检验ZO1-Cx43的假设 相互作用是调节细胞-细胞接触模式和迁移的机制的组成部分 通过确定：1.已知的刺激神经脊细胞运动的因素，是否影响 Cx43、ZO-1与其他缝隙连接蛋白相互作用的体外研究 ZO1-Cx43相互作用足以扰乱神经脊细胞的速率和方向性的调节 3.ZO1-Cx43相互作用是否对神经脊细胞的定向迁移是必需的 在活体内。这项研究将为胚胎神经脊迁移的分子调控提供新的视角。 以及人类先天缺陷的起源。；

Proper coordination of neural crest migration is an absolute requirement for normal cardiovascular development - most particularly outflow tract morphogenesis. The gap junction protein Cx43 has a key function in this directed migration of cardiac neural crest cells. The molecular mechanism by which Cx43 affects neural crest motility is unknown. The forces driving cell migration are generated largely by the dynamics of the actin cytoskeleton. We hypothesize that proteins mediating linkage between connexins and actin are candidates for regulators of cell motility. One molecule known to mediate interaction between connexins and actin is ZO1 - a PDZ protein related to the Drosophila tumor suppressor protein discs large. The Gourdie lab has shown that Cx43-ZO1 interaction is critical to developmental remodeling of the extentof gap junctional contact between myocardial cells. Furthermore, we have data showing that inhibition of ZO1- Cx43 interaction decreases neural crest outgrowth in vitro and reduces motility of fibroblasts and epithelial cells in cultured monolayers in a "scratch wound" migration assay. We will test the hypothesis that ZO1-Cx43 interaction is an integral part of a mechanism involved in regulation of cell-cell contact pattern and migration of neural crest cells by determining: 1. Whether factors known to stimulate neural crest cell motility, affect interactions between Cx43, ZO-1 and other connexin interacting proteins in vitro; 2. Whether inhibition of ZO1-Cx43 interaction is sufficient to disrupt regulation of the rate and directionality of neural crest cell migration in vitro; and 3. if ZO1-Cx43 interaction is necessary for the directed migration of neural crest cells in vivo. This study will provide new insight into molecular regulation of neural crest migration in the embryo and origins of birth defects in humans. ;