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通过体内电穿孔对成年小鼠神经发生微环境进行基因操作。

Genetic manipulation of adult mouse neurogenic niches by in vivo electroporation.

作者信息

Barnabé-Heider Fanie, Meletis Konstantinos, Eriksson Malin, Bergmann Olaf, Sabelström Hanna, Harvey Michael A, Mikkers Harald, Frisén Jonas

机构信息

Department of Cell and Molecular Biology, Medical Nobel Institute, SE-171 77 Stockholm, Sweden.

出版信息

Nat Methods. 2008 Feb;5(2):189-96. doi: 10.1038/nmeth.1174. Epub 2008 Jan 20.

Abstract

Targeted ectopic expression of genes in the adult brain is an invaluable approach for studying many biological processes. This can be accomplished by generating transgenic mice or by virally mediated gene transfer, but these methods are costly and labor intensive. We devised a rapid strategy that allows localized in vivo transfection of plasmid DNA within the adult neurogenic niches without detectable brain damage. Injection of plasmid DNA into the ventricular system or directly into the hippocampus of adult mice, followed by application of electrical current via external electrodes, resulted in transfection of neural stem or progenitor cells and mature neurons. We showed that this strategy can be used for both fate mapping and gain- or loss-of-function experiments. Using this approach, we identified an essential role for cadherins in maintaining the integrity of the lateral ventricle wall. Thus, in vivo electroporation provides a new approach to study the adult brain.

摘要

在成年大脑中进行基因的靶向异位表达是研究许多生物学过程的一种极有价值的方法。这可以通过生成转基因小鼠或通过病毒介导的基因转移来实现,但这些方法成本高昂且 labor intensive(此处可能有误,推测为labor-intensive,意为劳动密集型)。我们设计了一种快速策略,可在成年神经发生微环境中实现质粒DNA的局部体内转染,且不会造成可检测到的脑损伤。将质粒DNA注射到成年小鼠的脑室系统或直接注射到海马体中,随后通过外部电极施加电流,导致神经干细胞或祖细胞以及成熟神经元的转染。我们表明,该策略可用于命运图谱绘制以及功能获得或丧失实验。使用这种方法,我们确定了钙黏着蛋白在维持侧脑室壁完整性方面的重要作用。因此,体内电穿孔为研究成年大脑提供了一种新方法。

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