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

Neurosci Res. 2008 Apr;60(4):439-51. doi: 10.1016/j.neures.2007.12.011. Epub 2008 Jan 6.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • DNA, Complementary
  • DNA-Directed RNA Polymerases
  • Embryo, Mammalian
  • Gene Expression Profiling / methods*
  • Glutamate Decarboxylase / genetics
  • Glutamate Decarboxylase / metabolism*
  • Green Fluorescent Proteins / biosynthesis
  • Mice
  • Mice, Transgenic
  • Microarray Analysis / methods*
  • Neocortex / cytology
  • Neurons / metabolism*
  • Stem Cells / metabolism*
  • Viral Proteins
  • gamma-Aminobutyric Acid / genetics
  • gamma-Aminobutyric Acid / metabolism*

Substances

  • DNA, Complementary
  • Viral Proteins
  • Green Fluorescent Proteins
  • gamma-Aminobutyric Acid
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases
  • Glutamate Decarboxylase
  • glutamate decarboxylase 1