A transgene-assisted genetic screen identifies essential regulators of vascular development in vertebrate embryos
A transgene-assisted genetic screen identifies essential regulators of vascular development in vertebrate embryos
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DOI：
10.1016/j.ydbio.2007.03.526
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发表时间：
2007-07-01
影响因子：
2.7
通讯作者：
Stainier, Didier Y. R.
中科院分区：
文献类型：
--
作者：
Jin, Suk-Won;Herzog, Wiebke;Stainier, Didier Y. R.
哺乳动物发育过程中血管系统的形成对胚胎存活至关重要。此外，血管生长的不平衡有助于许多疾病的发病机制。我们对血管发育和血管生长的大部分理解来自于对Vegf信号通路的研究，以及最近对参与轴突引导的分子也调节血管模式的观察。为了采用一种公正而集中的方法来鉴定调节血管发育的新基因，我们在斑马鱼中进行了三步ENU突变筛选。我们首先从视觉上筛选活胚胎，评估主干血管（由血管生成）和分裂间血管（由血管生成）的血流量。将循环减少或无循环的胚胎固定并染色检测内源性碱性磷酸酶活性以显示血管形态。然后将所有假定的突变体交叉到Tg(flk1:EGFP)（s843）转基因背景中，以便对活胚胎和固定胚胎中的内皮细胞进行详细检查。我们筛选了4015个基因组，确定了30个影响血管发育各个方面的突变。具体来说，我们确定了3个基因（或基因座）调节内皮细胞的规范和/或分化，8个基因调节血管管和管腔形成，8个基因调节血管模式，1个基因调节血管重塑，完整性和维持。这些基因中只有4个先前与斑马鱼的血管发育有关，说明了这种聚焦筛选的价值。对新定义的基因座的分析将有助于更好地了解血管发育，并可能提供新的药物靶点来治疗与血管生长失调相关的多种病理。(c) 2007爱思唯尔公司版权所有。
Formation of a functional vasculature during mammalian development is essential for embryonic survival. In addition, imbalance in blood vessel growth contributes to the pathogenesis of numerous disorders. Most of our understanding of vascular development and blood vessel growth comes from investigating the Vegf signaling pathway as well as the recent observation that molecules involved in axon guidance also regulate vascular patterning. In order to take an unbiased, yet focused, approach to identify novel genes regulating vascular development, we performed a three-step ENU mutagenesis screen in zebrafish. We first screened live embryos visually, evaluating blood flow in the main trunk vessels, which form by vasculogenesis, and the intersomitic vessels, which form by angiogenesis. Embryos that displayed reduced or absent circulation were fixed and stained for endogenous alkaline phosphatase activity to reveal blood vessel morphology. All putative mutants were then crossed into the Tg(flk1:EGFP)(s843) transgenic background to facilitate detailed examination of endothelial cells in live and fixed embryos.We screened 4015 genomes and identified 30 mutations affecting various aspects of vascular development. Specifically, we identified 3 genes (or loci) that regulate the specification and/or differentiation of endothelial cells, 8 genes that regulate vascular tube and lumen fori-nation, 8 genes that regulate vascular patterning, and I I genes that regulate vascular remodeling, integrity and maintenance. Only 4 of these genes had previously been associated with vascular development in zebrafish illustrating the value of this focused screen. The analysis of the newly defined loci should lead to a greater understanding of vascular development and possibly provide new drug targets to treat the numerous pathologies associated with dysregulated blood vessel growth. (c) 2007 Elsevier Inc. All rights reserved.