Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors

被引:37
作者
Esumi, Shigeyuki [1 ]
Wu, Sheng-Xi [2 ,3 ]
Yanagawa, Yuchio [4 ,5 ]
Obata, Kunihiko [6 ]
Suaimoto, Yukihiko [7 ]
Tamamaki, Nobuald [1 ]
机构
[1] Kumamoto Univ, Grad Sch Med Sci, Dept Morphol Neural Sci, Kumamoto 8608556, Japan
[2] Fourth Mil Med Univ, Dept Anat, Xian 710032, Peoples R China
[3] Fourth Mil Med Univ, KK Leung Brain Res Ctr, Xian 710032, Peoples R China
[4] Gunma Univ, Grad Sch Med, Dept Genet & Behav Neurosci, Maebashi, Gunma 371, Japan
[5] SORST, Kawaguchi, Saitama, Japan
[6] RIKEN, BSI, Neuronal Circuit Mech Res Grp, Wako, Saitama, Japan
[7] Kyoto Univ, Grad Sch Pharmaceut Sci, Dept Physiol Chem, Kyoto, Japan
基金
日本学术振兴会;
关键词
GABA neuron; GFP; microarray; profiling; single cell;
D O I
10.1016/j.neures.2007.12.011
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
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.
引用
收藏
页码:439 / 451
页数:13
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