Method for single-cell microarray analysis and application to gene-expression profiling of GABAergic neuron progenitors.
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.
Esumi
Shigeyuki
S
Department of Morphological Neural Science, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.
Wu
Sheng-Xi
SX
Yanagawa
Yuchio
Y
Obata
Kunihiko
K
Sugimoto
Yukihiko
Y
Tamamaki
Nobuaki
N
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*
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