Role of Plcg1 in Vegf signaling

描述（由申请人提供）：在正常和疾病状态下控制血管形成的信号是相似的，并且在整个脊椎动物进化过程中是保守的。这使得可以使用遗传上易于处理的动物模型，例如斑马鱼，来解剖和分析这些信号。我们发现血管内皮生长因子（VEGF）是斑马鱼动脉形成和动脉内皮细胞分化所必需的特异性因子。由于VEGF已成为人类疾病中血管治疗操作的靶标，因此更好地理解VEGF驱动动脉发育的机制具有临床意义。我们的工作证明了磷脂酶C γ-1（Plcg 1）在VEGF介导的动脉发育过程中的作用，并表明其他信号也聚集在Plcg 1上以驱动这一过程。在这个建议中，我们将确定与Plcg 1在动脉发育过程中相互作用的分子，首先通过体内结构/功能分析确定动脉发育所需的Plcg 1结构域。基于这些结果，我们将确定与Plcg 1相互作用的分子，并确定它们在动脉发育过程中的功能。我们还将确定在动脉发育过程中影响VEGF和Plcg 1信号的新斑马鱼突变体。为此，我们将使用荧光标记血管的转基因斑马鱼进行遗传筛选，以便于识别突变表型。最后，我们将描述节段动脉突变表型，并确定候选基因负责这些表型。这些研究将传统的生物化学技术与通过使用斑马鱼系统获得的强大遗传工具相结合，并将提供一种全面的方法来更好地了解VEGF和Plcg 1如何控制动脉发育。由于这些信号传导机制是进化保守的，我们在该提案中确定的分子代表了病理血管形成的临床操作的可能靶点。

DESCRIPTION (provided by applicant): The signals that govern blood vessel formation in both normal and disease states are similar and have been conserved throughout vertebrate evolution. This enables the use of a genetically tractable animal model, such as the zebrafish, to dissect and analyze these signals. We have found that vascular endothelial growth factor (VEGF) is required specifically for the formation of arteries and the differentiation of arterial endothelial cells in zebrafish. Since VEGF has become a target for therapeutic manipulation of blood vessels in human diseases, it is clinically relevant to better understand the mechanism by which VEGF drives artery development. Our work demonstrates a role for phospholipase C gamma-1 (Plcg1) during VEGF-mediated artery development and suggests that other signals also converge on Plcg1 to drive this process. In this proposal, we will identify molecules that interact with Plcg1 during artery development by first identifying domains in Plcg1 that are required for artery development through in vivo structure/function analysis. Based on these results, we will identify molecules that interact with Plcg1 and determine their function during artery development. We will also identify new zebrafish mutants that affect VEGF and Plcg1 signaling during artery development. For this purpose we will perform a genetic screen using transgenic zebrafish with fluorescently labeled blood vessels that allow easy identification of mutant phenotypes. Finally, we will characterize segmental artery mutant phenotypes and identify candidate genes responsible for these phenotypes. These studies integrate traditional biochemical techniques with the powerful genetic tools available through the use of the zebrafish system and will provide a comprehensive approach to better understand how Vegf and Plcg1 govern artery development. Since these signaling mechanisms are evolutionary conserved, the molecules we identify in this proposal represent possible targets for clinical manipulation of pathological blood vessel formation.