斑马鱼成血血管细胞形成及其分子机制

项目名称： 斑马鱼成血血管细胞形成及其分子机制

项目编号： No.31271549

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 熊敬维

作者单位： 北京大学

项目金额： 90万元

中文摘要： 成血血管细胞被认为是血管内皮细胞和造血祖细胞的共同前体细胞，但它的起源及发育的分子机制很不清楚。前人的工作支持成血血管细胞来自斑马鱼原肠胚的腹侧边缘细胞，而我们观察发现flk1阳性细胞在背侧更加的富集并参与血管内皮、心脏内皮和脊索形成，由此计划重新用光转换荧光蛋白kaede和caged fluorescein dextran的标记方法对成血血管细胞的命运进行系统分析；并解析flk1启动子，发现关键序列，通过候选信号通路和酵母单杂交的方法鉴定出调控flk1在原肠期表达的关键因子；cloche斑马鱼突变体有成血血管细胞缺陷，计划利用mRNA深度测序鉴定、深入研究cloche调控并参与成血血管细胞发育的重要基因。总之本课题预期发现成血血管细胞的新起源、并揭示其发育的分子机理，有望揭示cloche突变体的遗传基础，在血管发育生物学基础研究及相关疾病临床应用都有重要意义。

中文关键词： 成血血管细胞；侧板中胚层；kdra；wnt；血管生成

英文摘要： It is well perceived that both angioblasts and hematopoietic stem cells are derived from hemangioblasts, however it remains largely unknown what are cellular and molecular mechanisms underlying hemangioblast development. Hemangioblast is shown to derive from the ventrolateral mesoderm during gastrulation in zebrafish. Our observation suggests the Flk1-positive dorsal mesodermal cells give rise to vascular endothelial cells, endocardial cells and the notochord, supporting a new source of hemangioblasts at the dorsal shield during zebrafish gastrulation. Here we plan to carry out a systematic cell lineage mapping of the Flk1-positive dorsal mesoderm by using photoconvertible Kaede and caged fluorescein dextran methods. By applying Tg(flk1:kaede) as the dorsal mesoderm reporter and small molecule inhibitors for known signaling pathways, we plan to determine which signaling pathways are required for generation of the Flk1-positive dorsal mesoderm. Aternatively, by utilizing a critical enhancer of the flk1 which directs the EGFP reporter gene expression at the shield and the Matchmaker One-hybrid system, we plan to isolate and characterize transcriptional factors that bind to the flk1 enhancer. Finally, taking advantage of a hemangioblast-deficient mutant cloche, we plan to isolate and analyze cloche-regulated genes

英文关键词： Angioblast；lateral plate mesoderm；kdra；wnt；vasculogenesis