Role of Plcg1 in Vegf signaling

Plcg1 在 Vegf 信号传导中的作用

• 批准号: 6872703
• 负责人: NATHAN D LAWSON
• 金额: $ 32万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-15 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6872703
• 关键词: alternatives to animals in research; angiogenesis; artery; biological signal transduction; cell differentiation; enzyme activity; fluorescent dye /probe; genetic markers; genetic screening; genetically modified animals; intermolecular interaction; mutant; phospholipase C; protein protein interaction; protein structure function; protein tyrosine kinase; vascular endothelial growth factors; vascular endothelium; zebrafish

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.

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一直是保守的。这使得能够使用遗传上易于处理的动物模型（例如斑马鱼）来剖析和分析这些信号。我们发现斑马鱼的动脉形成和动脉内皮细胞的分化特别需要血管内皮生长因子（VEGF）。由于 VEGF 已成为人类疾病中血管治疗操作的靶点，因此更好地了解 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 gamma-1 (Plcg1) 在 VEGF 介导的动脉发育过程中的作用，并表明其他信号也汇聚在 Plcg1 上来驱动这一过程。在本提案中，我们将首先通过体内结构/功能分析识别动脉发育所需的 Plcg1 结构域，从而识别动脉发育过程中与 Plcg1 相互作用的分子。基于这些结果，我们将鉴定与 Plcg1 相互作用的分子并确定它们在动脉发育过程中的功能。我们还将鉴定在动脉发育过程中影响 VEGF 和 Plcg1 信号传导的新斑马鱼突变体。为此，我们将使用带有荧光标记血管的转基因斑马鱼进行遗传筛选，以便轻松识别突变表型。最后，我们将描述节段动脉突变表型的特征，并确定导致这些表型的候选基因。这些研究将传统的生化技术与通过使用斑马鱼系统提供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 Plcg1 如何控制动脉发育。由于这些信号机制在进化上是保守的，因此我们在本提案中确定的分子代表了病理性血管形成的临床操作的可能目标。