Kamme Fredrik, Salunga Ranelle, Yu Jingxue, Tran Da-Thao, Zhu Jessica, Luo Lin, Bittner Anton, Guo Hong-Qing, Miller Nancy, Wan Jackson, Erlander Mark
Johnson & Johnson Pharmaceutical Research and Development LLC, San Diego, California 92121, USA.
J Neurosci. 2003 May 1;23(9):3607-15. doi: 10.1523/JNEUROSCI.23-09-03607.2003.
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.
激光捕获显微切割技术与微阵列相结合,能够对选定细胞中的数千个基因进行表达分析。在此,我们描述了一种利用激光捕获、T7 RNA扩增和cDNA微阵列分析相结合的方法,对大鼠海马体CA1神经元进行单细胞基因表达谱分析。随后对微阵列数据进行的聚类分析确定了两种不同的细胞类型:锥体神经元和中间神经元。聚类分析还揭示了锥体神经元之间的差异,这表明即使是体内的单一细胞类型,在基因表达水平上也不是一个同质的细胞群体。微阵列数据通过定量RT-PCR和原位杂交得到了证实。我们还报告了这种方法组合的可重复性和灵敏度。单细胞基因表达谱分析为解决哺乳动物大脑的复杂性提供了一个强大的工具。