Notch-Gridlock Signaling in Vascular Specification

描述(申请人提供)：血管发育需要一系列复杂的事件，在此过程中内皮细胞从血管母细胞前体细胞分化，并聚集到背主动脉和主静脉，构成原始的循环系统。动脉和静脉在形态、功能和分子上是不同的。在血管形成的早期阶段，这种区别是如何在细胞和分子水平上建立起来的，在很大程度上是未知的。 斑马鱼研究的新遗传证据表明，Notch信号转导通路是维管指定所必需的。GRIDLOCK基因编码分裂相关bHLH蛋白的增强子，介导Notch信号促进胚胎动脉的发育。该研究方案解释了如何在斑马鱼中使用细胞和遗传分析来了解缺口-僵局途径在调节血管母细胞的动脉/静脉内皮细胞命运中的作用。 特别是，这个建议首先旨在通过跟踪血管母细胞的迁移来确定潜在的细胞-细胞相互作用。接下来，实验旨在研究Notch信号成分在血管发育中的表达模式，以及确定不能正常传递Notch信号的单个血管母细胞的命运。最后的实验是为了测试网格锁定通过负反馈环路中介Notch信号的机制。 由于血管发育的基本原理在脊椎动物和哺乳动物之间是保守的，了解斑马鱼的缺口-栅格信号通路将有助于深入了解血管发育的机制，并促进我们对许多人类动脉疾病的病因的理解。

DESCRIPTION (provided by applicant): Vascular development requires a complex series of events during which endothelial cells differentiate from angioblast progenitors and assemble into the dorsal aorta and the cardinal vein, comprising the original circulatory loop. Arteries and veins are morphologically, functionally and molecularly different. How this distinction is established at cellular and molecular levels in the early stage of vasculogenesis is largely unknown. New genetic evidence from zebra fish studies suggests that the Notch signal transduction pathway is required for vascular specification. The gridlock gene, encoding an Enhancer of Split-related bHLH protein, mediates Notch signaling to promote development of the embryonic arteries. The research proposal explains how cellular and genetic analysis are used in zebra fish to understand the role of notch-gridlock pathway in regulating arterial/venous endothelial fate of angioblasts. Specially, this proposal first aims to identify potential cell-cell interactions by following angioblast migration. Next, experiments are designed to examine expression pattern of Notch signaling components in vascular development, as well as to determine the fate of individual angioblasts that are incapable of normal Notch signaling. The final experiments are proposed to test the mechanism by which gridlock mediates Notch signaling via a negative feedback loop. As fundamentals of vascular development are conserved between vertebrates and mammals, understanding the notch-gridlock signaling pathway in zebrafish will provide insights into mechanisms of vascular development as well as advance our understanding of etiologies of many human arterial diseases.