Patch-Seq Protocol to Analyze the Electrophysiology, Morphology and Transcriptome of Whole Single Neurons Derived From Human Pluripotent Stem Cells.
Patch-Seq Protocol to Analyze the Electrophysiology, Morphology and Transcriptome of Whole Single Neurons Derived From Human Pluripotent Stem Cells.
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DOI：
10.3389/fnmol.2018.00261
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发表时间：
2018
影响因子：
4.8
通讯作者：
Bardy C
中科院分区：
文献类型：
--
作者：
van den Hurk M;Erwin JA;Yeo GW;Gage FH;Bardy C
人类大脑由大约1710亿个异质细胞单位（860亿个神经元和850亿个非神经元胶质细胞）的复杂组装体组成。对脑细胞的全面描述对于了解健康和疾病中的神经系统是必要的。最近，基因组学的进步已经允许精确分析单细胞的全转录组（scRNA-seq）。我们已经建立在这样的技术进步，联合收割机scRNA-seq与膜片钳电生理记录和体外单个人类神经元的形态学分析相结合。这种新的强大方法，称为Patch-seq，能够对神经元进行彻底的多模态分析，并使我们能够揭示功能特性，形态和基因表达之间的联系。在这里，我们提出了一个详细的Patch-seq协议，用于从体外神经元培养物中分离单个神经元。我们已经用来自健康受试者的胚胎和诱导多能干细胞（ESC/iPSC）产生的人类神经元验证了Patch-seq全转录组分析方法，但该程序可应用于体外任何类型的细胞。Patch-seq可用于体外神经元，以深入分析细胞类型和状态，从而揭示神经元多样性的人类分子基础，并研究大脑疾病的细胞机制。
The human brain is composed of a complex assembly of about 171 billion heterogeneous cellular units (86 billion neurons and 85 billion non-neuronal glia cells). A comprehensive description of brain cells is necessary to understand the nervous system in health and disease. Recently, advances in genomics have permitted the accurate analysis of the full transcriptome of single cells (scRNA-seq). We have built upon such technical progress to combine scRNA-seq with patch-clamping electrophysiological recording and morphological analysis of single human neurons in vitro. This new powerful method, referred to as Patch-seq, enables a thorough, multimodal profiling of neurons and permits us to expose the links between functional properties, morphology, and gene expression. Here, we present a detailed Patch-seq protocol for isolating single neurons from in vitro neuronal cultures. We have validated the Patch-seq whole-transcriptome profiling method with human neurons generated from embryonic and induced pluripotent stem cells (ESCs/iPSCs) derived from healthy subjects, but the procedure may be applied to any kind of cell type in vitro. Patch-seq may be used on neurons in vitro to profile cell types and states in depth to unravel the human molecular basis of neuronal diversity and investigate the cellular mechanisms underlying brain disorders.
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影响因子：
46.9
作者：
Fuzik J;Zeisel A;Máté Z;Calvigioni D;Yanagawa Y;Szabó G;Linnarsson S;Harkany T
通讯作者：
Harkany T
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
11
作者：
Bardy, C.;van den Hurk, M.;Kakaradov, B.;Erwin, J. A.;Jaeger, B. N.;Hernandez, R. V.;Eames, T.;Paucar, A. A.;Gorris, M.;Marchand, C.;Jappelli, R.;Barron, J.;Bryant, A. K.;Kellogg, M.;Lasken, R. S.;Rutten, B. P. F.;Steinbusch, H. W. M.;Yeo, G. W.;Gage, F. H.
通讯作者：
Gage, F. H.
影响因子：
--
作者：
Boyer, Leah F;Campbell, Benjamin;Gage, Fred H
通讯作者：
Gage, Fred H
影响因子：
46.9
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
通讯作者：
Tolias AS