title: Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors.
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.
authors.affiliation: Department of Morphological Neural Science, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Esumi
authors.forename: Sheng-Xi
authors.initials: SX
authors.lastname: Wu
authors.forename: Yuchio
authors.initials: Y
authors.lastname: Yanagawa
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Obata
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Sugimoto
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Tamamaki
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018076:DNA, Complementary; D012321:DNA-Directed RNA Polymerases; D004622:Embryo, Mammalian; D020869:Gene Expression Profiling / Q000379:methods*; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D051379:Mice; D008822:Mice, Transgenic; D046228:Microarray Analysis / Q000379:methods*; D019579:Neocortex / Q000166:cytology; D009474:Neurons / Q000378:metabolism*; D013234:Stem Cells / Q000378:metabolism*; D014764:Viral Proteins; D005680:gamma-Aminobutyric Acid / Q000235:genetics / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D005680:gamma-Aminobutyric Acid; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D005968:Glutamate Decarboxylase; C402565:glutamate decarboxylase 1