FUNCTION OF CX43-INTERACTING PROTEINS IN NEURAL CREST MIGRATION

神经嵴迁移的适当协调是正常心血管疾病的绝对要求。 发育-最特别是流出道形态发生。差距连接蛋白Cx43是一个关键 在心脏神经嵴细胞的定向迁移中发挥作用。Cx43的分子机制 影响神经嵴运动的原因尚不清楚驱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连接的蛋白质， 肌动蛋白是细胞运动调节剂的候选者。已知一种分子介导 连接蛋白和肌动蛋白是ZO 1-PDZ蛋白相关的果蝇肿瘤抑制蛋白盘大。 Gourdie实验室已经表明，Cx43-ZO 1相互作用对发育重塑的程度至关重要。 心肌细胞之间的缝隙连接。此外，我们有数据表明，抑制ZO 1- Cx43相互作用降低体外神经嵴生长并降低成纤维细胞和上皮细胞的运动性 在“划痕”迁移试验中，培养单层中的细胞。我们将检验ZO 1-Cx43 相互作用是涉及调节细胞-细胞接触模式和迁移机制的组成部分 的神经嵴细胞通过确定：1.是否已知刺激神经嵴细胞运动的因素， Cx43、ZO-1与其他连接蛋白相互作用蛋白的体外相互作用; 2.的抑制是否 ZO 1-Cx43相互作用足以破坏神经嵴细胞的速率和方向性调节 体外迁移; 3.如果ZO 1-Cx43相互作用是神经嵴细胞定向迁移所必需的， in vivo.这项研究将为胚胎神经嵴迁移的分子调控提供新的见解 和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. ;