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Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors |
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| Authors: | Esumi Shigeyuki Wu Sheng-Xi Yanagawa Yuchio Obata Kunihiko Sugimoto Yukihiko Tamamaki Nobuaki |
| Institution: | aDepartment of Morphological Neural Science, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan bDepartment of Anatomy and K.K. Leung Brain Research Centre, Fourth Military Medical University, Xi’an, PR China cDepartment of Genetic and Behavioral Neuroscience, Graduate School of Medicine, Gunma University, Maebashi, Japan dSORST, Kawaguchi, Japan eNeuronal Circuit Mechanism Research Group, RIKEN, BSI, Wako, Japan fDepartment of Physiological Chemistry, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan |
| Abstract: | 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. |
| Keywords: | GABA neuron GFP Microarray Profiling Single cell |
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