The Journal of Neuroscience, May 1, 2003, 23(9):3607
Single-Cell Microarray Analysis in Hippocampus CA1: Demonstration
and Validation of Cellular Heterogeneity
Fredrik
Kamme1,
Ranelle
Salunga2,
Jingxue
Yu1,
Da-Thao
Tran1,
Jessica
Zhu1,
Lin
Luo1,
Anton
Bittner1,
Hong-Qing
Guo1,
Nancy
Miller3,
Jackson
Wan1, and
Mark
Erlander2
1 Johnson & Johnson Pharmaceutical Research and
Development LLC, San Diego, California 92121, 2 Arcturus Applied Genomics, Carlsbad, California 92008, and 3 OmniViz, Maynard, Massachusetts 01754
Laser capture microdissection in combination with microarrays
allows for the expression analysis of thousands of genes in selected
cells. Here we describe single-cell gene expression profiling of CA1
neurons in the rat hippocampus using a combination of laser capture, T7
RNA amplification, and cDNA microarray analysis. Subsequent cluster
analysis of the microarray data identified two different cell types:
pyramidal neurons and an interneuron. Cluster analysis also revealed
differences among the pyramidal neurons, indicating that even a single
cell type in vivo is not a homogeneous population of cells at the gene expression level. Microarray data were confirmed by quantitative RT-PCR and in situ hybridization. We
also report on the reproducibility and sensitivity of this combination
of methods. Single-cell gene expression profiling offers a powerful tool to tackle the complexity of the mammalian brain.
Key words:
laser capture microdissection; gene expression
profiling; RNA amplification; single cell; microarray; cell type
definition
Copyright © 2003 Society for Neuroscience 0270-6474/03/2393607-09$05.00/0