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利用 piggyBac 系统和子宫内电穿孔在小鼠大脑中进行神经胶质细胞类型特异性基因表达。

Glial cell type-specific gene expression in the mouse cerebrum using the piggyBac system and in utero electroporation.

机构信息

Department of Medical Neuroscience, Graduate School of Medical Sciences, Kanazawa University, Takara-machi 13-1, Kanazawa, Ishikawa, 920-8640, Japan.

Center for Biomedical Research and Education, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, 920-8640, Japan.

出版信息

Sci Rep. 2021 Mar 1;11(1):4864. doi: 10.1038/s41598-021-84210-z.

Abstract

Glial cells such as astrocytes and oligodendrocytes play crucial roles in the central nervous system. To investigate the molecular mechanisms underlying the development and the biological functions of glial cells, simple and rapid techniques for glial cell-specific genetic manipulation in the mouse cerebrum would be valuable. Here we uncovered that the Gfa2 promoter is suitable for selective gene expression in astrocytes when used with the piggyBac system and in utero electroporation. In contrast, the Blbp promoter, which has been used to induce astrocyte-specific gene expression in transgenic mice, did not result in astrocyte-specific gene expression. We also identified the Plp1 and Mbp promoters could be used with the piggyBac system and in utero electroporation to induce selective gene expression in oligodendrocytes. Furthermore, using our technique, neuron-astrocyte or neuron-oligodendrocyte interactions can be visualized by labeling neurons, astrocytes and oligodendrocytes differentially. Our study provides a fundamental basis for specific transgene expression in astrocytes and/or oligodendrocytes in the mouse cerebrum.

摘要

神经胶质细胞,如星形胶质细胞和少突胶质细胞,在中枢神经系统中发挥着至关重要的作用。为了研究神经胶质细胞发育和生物学功能的分子机制,在小鼠大脑中进行简单快速的神经胶质细胞特异性基因操作的技术将是非常有价值的。在这里,我们发现 Gfa2 启动子与 piggyBac 系统和体内电穿孔一起使用时,适合于星形胶质细胞的选择性基因表达。相比之下,Blbp 启动子已被用于诱导转基因小鼠中星形胶质细胞特异性基因表达,但不能导致星形胶质细胞特异性基因表达。我们还鉴定出 Plp1 和 Mbp 启动子可以与 piggyBac 系统和体内电穿孔一起使用,以诱导少突胶质细胞的选择性基因表达。此外,使用我们的技术,可以通过标记神经元、星形胶质细胞和少突胶质细胞的差异来可视化神经元-星形胶质细胞或神经元-少突胶质细胞的相互作用。我们的研究为在小鼠大脑中特异性地在星形胶质细胞和/或少突胶质细胞中转基因表达提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b637/7921133/9e2071612317/41598_2021_84210_Fig1_HTML.jpg

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