Nuclear RNA-seq of single neurons reveals molecular signatures of activation.
Nuclear RNA-seq of single neurons reveals molecular signatures of activation.
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DOI：
10.1038/ncomms11022
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发表时间：
2016-04-19
影响因子：
16.6
通讯作者：
Gage FH
中科院分区：
文献类型：
--
作者：
Lacar B;Linker SB;Jaeger BN;Krishnaswami SR;Barron JJ;Kelder MJE;Parylak SL;Paquola ACM;Venepally P;Novotny M;O'Connor C;Fitzpatrick C;Erwin JA;Hsu JY;Husband D;McConnell MJ;Lasken R;Gage FH
单细胞测序方法已成为鉴定特定脑区内异质细胞类型的有力工具。应用单细胞技术来研究激活的神经元的转录组可以提供与对给定经历的不同神经元反应相关的分子动力学的见解。通过对常见的全细胞和单核RNA测序（snRNA-seq）方法的评估，我们发现snRNA-seq忠实地再现了与经验驱动的活性诱导相关的转录模式，包括立即早期基因（IEGs），如Fos，Arc和Egr 1。小鼠齿状颗粒细胞的SnRNA-seq揭示了短暂的新环境暴露后激活的神经元转录组的大规模变化，包括MAPK通路基因的诱导。此外，我们观察到一个连续的激活状态，揭示了一个pseudotemporal模式的激活基因表达单独。总之，激活神经元的snRNA-seq能够检查IEG以外的基因表达，从而对体内神经元激活模式有了新的见解。 与体内神经元激活模式相关的分子动力学尚不清楚。Lacar等人对暴露于新环境的小鼠的海马神经元进行单核RNA测序，并确定与经验相关的单个神经元中的大规模转录组变化。
Single-cell sequencing methods have emerged as powerful tools for identification of heterogeneous cell types within defined brain regions. Application of single-cell techniques to study the transcriptome of activated neurons can offer insight into molecular dynamics associated with differential neuronal responses to a given experience. Through evaluation of common whole-cell and single-nuclei RNA-sequencing (snRNA-seq) methods, here we show that snRNA-seq faithfully recapitulates transcriptional patterns associated with experience-driven induction of activity, including immediate early genes (IEGs) such as Fos, Arc and Egr1. SnRNA-seq of mouse dentate granule cells reveals large-scale changes in the activated neuronal transcriptome after brief novel environment exposure, including induction of MAPK pathway genes. In addition, we observe a continuum of activation states, revealing a pseudotemporal pattern of activation from gene expression alone. In summary, snRNA-seq of activated neurons enables the examination of gene expression beyond IEGs, allowing for novel insights into neuronal activation patterns in vivo. The molecular dynamics associated with neuronal activation patterns in vivo are unclear. Lacar et al. perform single-nuclei RNA-sequencing of hippocampal neurons from mice exposed to a novel environment, and identify large-scale transcriptome changes in individual neurons associated with the experience.