Single-cell microarray analysis in hippocampus CA1: demonstration and validation of cellular heterogeneity.
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.
Kamme
Fredrik
F
Johnson & Johnson Pharmaceutical Research and Development LLC, San Diego, California 92121, USA. fkamme@prdus.jnj.com
Salunga
Ranelle
R
Yu
Jingxue
J
Tran
Da-Thao
DT
Zhu
Jessica
J
Luo
Lin
L
Bittner
Anton
A
Guo
Hong-Qing
HQ
Miller
Nancy
N
Wan
Jackson
J
Erlander
Mark
M
D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D002452:Cell Count; D016000:Cluster Analysis; D005260:Female; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D017403:In Situ Hybridization; D007395:Interneurons / Q000737:chemistry / Q000378:metabolism; D007834:Lasers; D009474:Neurons / Q000737:chemistry / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D017966:Pyramidal Cells / Q000737:chemistry / Q000378:metabolism; D016372:RNA, Antisense / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D051381:Rats; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D016372:RNA, Antisense; D012333:RNA, Messenger