Electrophysiological, transcriptomic and morphologic profiling of single neurons using Patch-seq.
Electrophysiological, transcriptomic and morphologic profiling of single neurons using Patch-seq.
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DOI：
10.1038/nbt.3445
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发表时间：
2016-02
影响因子：
46.9
通讯作者：
Tolias AS
中科院分区：
文献类型：
--
作者：
Cadwell CR;Palasantza A;Jiang X;Berens P;Deng Q;Yilmaz M;Reimer J;Shen S;Bethge M;Tolias KF;Sandberg R;Tolias AS
尽管哺乳动物的新皮层对于复杂的认知过程很重要，但我们仍然缺乏对其细胞成分的全面描述。为了改善神经元细胞类型的分类和单个神经元的功能表征，我们提出了Patch-seq，一种结合全细胞电生理膜片钳记录，单细胞RNA测序和形态学表征的方法。电生理学表征后，通过膜片钳移液管吸出细胞内容物并制备用于RNA测序。使用这种方法，我们产生的58个新皮层细胞的电生理和分子特征，并表明基因表达模式可以用来推断的形态和生理特性，如轴突分支和动作电位振幅的个别神经元。我们的研究结果揭示了神经元多样性的分子基础，并表明Patch-seq可以促进神经系统中细胞类型的分类。
Despite the importance of the mammalian neocortex for complex cognitive processes, we still lack a comprehensive description of its cellular components. To improve the classification of neuronal cell types and the functional characterization of single neurons, we present Patch-seq, a method that combines whole-cell electrophysiological patch-clamp recordings, single-cell RNA-sequencing and morphological characterization. Following electrophysiological characterization, cell contents are aspirated through the patch-clamp pipette and prepared for RNA-sequencing. Using this approach, we generate electrophysiological and molecular profiles of 58 neocortical cells and show that gene expression patterns can be used to infer the morphological and physiological properties such as axonal arborization and action potential amplitude of individual neurons. Our results shed light on the molecular underpinnings of neuronal diversity and suggest that Patch-seq can facilitate the classification of cell types in the nervous system.