Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors
Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors
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DOI：
10.1016/j.neures.2007.12.011
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发表时间：
2008-04-01
影响因子：
2.9
通讯作者：
Tamamaki, Nobuald
中科院分区：
文献类型：
--
作者：
Esumi, Shigeyuki;Wu, Sheng-Xi;Tamamaki, Nobuald
哺乳动物的中枢神经系统由各种类型的神经元和神经胶质细胞组成。要研究单个细胞的功能和发育，需要在单细胞水平上进行基因表达分析。在这里，我们开发了一种基于微阵列的单细胞基因表达谱方法，并对GABA能神经元前体细胞进行了测试。分离GAD67-GFP基因敲除小鼠的新皮质，提取GFP阳性细胞，收集单个GABA能神经元祖细胞。在优化的条件下，将Super SMART PCR和T7RNA聚合酶扩增相结合，从单细胞中扩增互补DNA。对cRNA进行微阵列杂交和分析，得到了可靠和重复性较好的结果。(C)2008年爱思唯尔爱尔兰有限公司和日本神经科学学会。版权所有。
The mammalian central nervous system is populated with various types of neurons and glia. To investigate the functions and development of individual cells requires gene-expression analysis at the single-cell level. Here, we developed a microarray-based method for the gene-expression profiling of single cells and tested it for GABAergic neuron progenitors. Single GABAergic neuron progenitors were collected from the neocortex of GAD67-GFP knock-in mice by dissociation followed by the aspiration of GFP-positive cells. Complementary DNA from the single cells was amplified by a method in which Super SMART PCR and T7 RNA polymerase amplification were combined at a optimized condition. The cRNA was subjected to microarray hybridization and analysis, which yielded reliable and reproducible results. (C) 2008 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.