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在脊索中从Foxa2和T位点产生表达核增强型绿色荧光蛋白和他莫昔芬诱导型Cre重组酶的敲入小鼠。

Generation of knock-in mice that express nuclear enhanced green fluorescent protein and tamoxifen-inducible Cre recombinase in the notochord from Foxa2 and T loci.

作者信息

Imuta Yu, Kiyonari Hiroshi, Jang Chuan-Wei, Behringer Richard R, Sasaki Hiroshi

机构信息

Department of Cell Fate Control, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.

出版信息

Genesis. 2013 Mar;51(3):210-8. doi: 10.1002/dvg.22376. Epub 2013 Feb 25.

Abstract

The node and the notochord are important embryonic signaling centers that control embryonic pattern formation. Notochord progenitor cells present in the node and later in the posterior end of the notochord move anteriorly to generate the notochord. To understand the dynamics of cell movement during notochord development and the molecular mechanisms controlling this event, analyses of cell movements using time-lapse imaging and conditional manipulation of gene activities are required. To achieve this goal, we generated two knock-in mouse lines that simultaneously express nuclear enhanced green fluorescent protein (EGFP) and tamoxifen-inducible Cre, CreER(T2) , from two notochord gene loci, Foxa2 and T (Brachury). In Foxa2(nEGFP-CreERT2/+) and T(nEGFP-CreERT2/+) embryos, nuclei of the Foxa2 or T-expressing cells, which include the node, notochord, and endoderm (Foxa2) or wide range of posterior mesoderm (T), were labeled with EGFP at intensities that can be used for live imaging. Cre activity was also induced in cells expressing Foxa2 and T 1 day after tamoxifen administration. These mice are expected to be useful tools for analyzing the mechanisms of notochord development.

摘要

节点和脊索是控制胚胎模式形成的重要胚胎信号中心。存在于节点以及后来脊索后端的脊索祖细胞向前移动以形成脊索。为了解脊索发育过程中细胞运动的动态变化以及控制这一事件的分子机制,需要使用延时成像技术分析细胞运动,并对基因活性进行条件性操纵。为实现这一目标,我们构建了两个敲入小鼠品系,它们分别从两个脊索基因位点Foxa2和T(Brachury)同时表达核增强型绿色荧光蛋白(EGFP)和他莫昔芬诱导型Cre,即CreER(T2)。在Foxa2(nEGFP-CreERT2/+)和T(nEGFP-CreERT2/+)胚胎中,表达Foxa2或T的细胞(包括节点、脊索和内胚层(Foxa2)或广泛的后中胚层(T))的细胞核被EGFP标记,其强度可用于实时成像。在给予他莫昔芬1天后,表达Foxa2和T的细胞中也诱导了Cre活性。这些小鼠有望成为分析脊索发育机制的有用工具。

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