Tietjen Ian, Rihel Jason M, Cao Yanxiang, Koentges Georgy, Zakhary Lisa, Dulac Catherine
Howard Hughes Medical Institute, Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Neuron. 2003 Apr 24;38(2):161-75. doi: 10.1016/s0896-6273(03)00229-0.
The extraordinary cellular heterogeneity of the mammalian nervous system has largely hindered the molecular analysis of neuronal identity and diversity. In order to uncover mechanisms involved in neuronal differentiation and diversification, we have monitored the expression profiles of individual neurons and progenitor cells collected from dissociated tissue or captured from intact slices. We demonstrate that this technique provides a sensitive and reproducible representation of the single-cell transcriptome. In the olfactory system, hundreds of transcriptional differences were identified between olfactory progenitors and mature sensory neurons, enabling us to define the large variety of signaling pathways expressed by individual progenitors at a precise developmental stage. Finally, we show that regional differences in gene expression can be predicted from transcriptional analysis of single neuronal precursors isolated by laser capture from defined areas of the developing brain.
哺乳动物神经系统中细胞的非凡异质性在很大程度上阻碍了对神经元特性和多样性的分子分析。为了揭示神经元分化和多样化所涉及的机制,我们监测了从解离组织中收集或从完整切片中捕获的单个神经元和祖细胞的表达谱。我们证明,这项技术能够灵敏且可重复地呈现单细胞转录组。在嗅觉系统中,我们鉴定出嗅觉祖细胞和成熟感觉神经元之间存在数百种转录差异,这使我们能够确定单个祖细胞在精确发育阶段所表达的多种信号通路。最后,我们表明,通过对从发育中大脑的特定区域经激光捕获分离出的单个神经元前体进行转录分析,可以预测基因表达的区域差异。