Generating and imaging multicolor Brainbow mice.
Generating and imaging multicolor Brainbow mice.
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DOI：
10.1101/pdb.top114
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发表时间：
2011-07
影响因子：
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通讯作者：
T. Weissman;J. Sanes;J. Lichtman;J. Livet
中科院分区：
文献类型：
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作者：
T. Weissman;J. Sanes;J. Lichtman;J. Livet
可视化紧密并列的细胞及其相互作用的精确形态可以提供大量信息，特别是在研究神经系统中神经元和神经胶质网络的复杂组织时。为此，人们可以使用光学方法来成像不同的标记物，这些标记物在感兴趣的细胞中有差异地分布，例如各种颜色的荧光蛋白（XFP）。Brainbow策略使用Cre/lox重组以从单个启动子在细胞群体中随机表达2至4个XFP。多个Brainbow转基因拷贝的整合导致这些XFP的组合表达，产生广泛的色调。在神经系统中，这样产生的标记可用于区分相邻的神经元或神经胶质细胞，并在追踪电路的同时验证神经元过程的身份。本文描述了Brainbow转基因和小鼠的产生，以及它们在固定样本中成像和数字重建神经细胞及其相互作用的用途。该方法也具有在其他组织和模式生物中的研究以及活体成像的潜力。
Visualizing the precise morphology of closely juxtaposed cells and their interactions can be highly informative, particularly when studying the complex organization of neuronal and glial networks in the nervous system. To this end, one can use optical approaches to image-distinct markers that are differentially distributed among the cells of interest, such as fluorescent proteins of various colors (XFPs). The Brainbow strategies use Cre/lox recombination to stochastically express two to four XFPs in a cellular population from a single promoter. Integration of multiple Brainbow transgene copies results in combinatorial expression of these XFPs, creating a wide range of hues. In the nervous system, the multicolor labeling thus generated can be used to distinguish adjacent neuronal or glial cells and to verify the identity of neuronal processes while tracing circuitry. This article describes the generation of Brainbow transgenes and mice as well as their use to image and digitally reconstruct nerve cells and their interactions in fixed samples. This method also holds potential for studies in other tissues and model organisms as well as live imaging in vivo.