Notch-Gridlock Signaling in Vascular Specification

描述（由申请人提供）：血管发育需要一系列复杂的事件，在此期间，内皮细胞从成血管细胞祖细胞分化并组装成背主动脉和主静脉，构成原始循环回路。动脉和静脉在形态、功能和分子上都不同。这种区别是如何在血管发生的早期阶段在细胞和分子水平上建立起来的，这在很大程度上是未知的。 来自斑马鱼研究的新遗传证据表明，Notch信号转导途径是血管特化所必需的。gridlock基因编码分裂相关bHLH蛋白的增强子，介导Notch信号传导以促进胚胎动脉的发育。该研究提案解释了如何在斑马鱼中使用细胞和遗传分析来了解notch-gridlock途径在调节成血管细胞的动脉/静脉内皮命运中的作用。 特别地，该提议首先旨在通过跟踪成血管细胞迁移来识别潜在的细胞-细胞相互作用。接下来，设计实验以检查Notch信号传导组分在血管发育中的表达模式，以及确定不能正常Notch信号传导的个体成血管细胞的命运。最后的实验提出了测试的机制，通过它gridlock介导的Notch信号通过负反馈回路。 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，因此了解斑马鱼中的Notch-gridlock信号通路将提供对血管发育机制的见解，并促进我们对许多人类动脉疾病病因的理解。

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.