Regulation of mesodermal progenitors in transgenic zebrafish

转基因斑马鱼中胚层祖细胞的调控

基本信息

批准号：
7877975
负责人：
David Kimelman
金额：
$ 32.01万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2012-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): A large body of evidence in vertebrates shows that the development of the anterior trunk and the rest of the body are differentially regulated. Previous studies using mutants, knockouts, morpholinos and mRNA injection have provided very important information on the role of signaling and transcription factors in establishing the embryonic vertebrate body plan during the early gastrula stages. However, because alterations in the patterning of the early gastrula embryo cause such severe embryological defects, it has been difficult to study later stages when most of the body develops. Using transgenic lines containing heat-shock inducible cell autonomous regulators of signaling and transcription, together with a new transgenic line we developed that allows us to temporally and specifically label the mesodermal progenitors, we now have the ability to determine how the mesodermal progenitors are regulated by signaling and T-box transcription factors to progressively differentiate during somitogenesis, a process that is essential for forming a normal body and is conserved in all vertebrates. The studies described here take advantage of the ability in zebrafish to analyze the mesodermal progenitors at the single-cell level in living embryos, using cell transplants to study experimentally cell-autonomously perturbed cells within a normal background. This novel approach will allow us to examine cell behaviors that are obscured when all of the embryonic cells have alterations in signaling or transcription. Studies, particularly in mammals, show that the mesodermal progenitors are a stem-cell like population, which normally has a defined lifetime. With recent advances in the technology to produce transgenic lines, combined with the ability to study progenitors in living embryos using fluorescent reporters, zebrafish provides an excellent model system for understanding how vertebrate stem cells are regulated in vivo. As stem cells have great promise for the treatment of many diseases, the studies described here will provide valuable information about the signaling networks and transcription factors that control stem cell maintenance and tissue formation. Studies, particularly in mammals, show that the mesodermal progenitors are a stem-cell like population, which normally has a defined lifetime. With recent advances in the technology to produce transgenic lines, combined with the ability to study progenitors in living embryos using fluorescent reporters, zebrafish provides an excellent model system for understanding how vertebrate stem cells are regulated in vivo. As stem cells have great promise for the treatment of many diseases, the studies described here will provide valuable information about the signaling networks and transcription factors that control stem cell maintenance and tissue formation.

描述（由申请人提供）：脊椎动物的大量证据表明，前躯干和身体其他部分的发育受到不同的调节。先前使用突变体、基因敲除、吗啉代和 mRNA 注射的研究提供了关于信号和转录因子在早期原肠胚阶段建立胚胎脊椎动物身体计划中的作用的非常重要的信息。然而，由于早期原肠胚胚胎模式的改变导致如此严重的胚胎缺陷，因此很难研究身体大部分发育的后期阶段。使用含有热休克诱导细胞信号和转录自主调节因子的转基因系，以及我们开发的新转基因系，使我们能够暂时和特异性地标记中胚层祖细胞，我们现在有能力确定中胚层祖细胞是如何被调节的信号传导和T-box转录因子在体节发生过程中逐渐分化，体节发生是形成正常身体所必需的过程，并且在所有脊椎动物中都是保守的。这里描述的研究利用斑马鱼在活胚胎的单细胞水平上分析中胚层祖细胞的能力，使用细胞移植来研究正常背景下实验性细胞自主扰动的细胞。这种新方法将使我们能够检查当所有胚胎细胞的信号或转录发生改变时被掩盖的细胞行为。研究，特别是在哺乳动物中，表明中胚层祖细胞是干细胞样群体，通常具有确定的寿命。随着转基因系技术的最新进展，再加上使用荧光报告基因研究活体胚胎中祖细胞的能力，斑马鱼为了解脊椎动物干细胞在体内的调节方式提供了一个极好的模型系统。由于干细胞在治疗许多疾病方面有着巨大的前景，这里描述的研究将提供有关控制干细胞维持和组织形成的信号网络和转录因子的宝贵信息。研究，特别是在哺乳动物中，表明中胚层祖细胞是干细胞样群体，通常具有确定的寿命。随着转基因系技术的最新进展，再加上使用荧光报告基因研究活体胚胎中祖细胞的能力，斑马鱼为了解脊椎动物干细胞在体内的调节方式提供了一个极好的模型系统。由于干细胞在治疗许多疾病方面有着巨大的前景，这里描述的研究将提供有关控制干细胞维持和组织形成的信号网络和转录因子的宝贵信息。