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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


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