Correlating Anatomy and Function with Gene Expression in Individual Neurons by Combining in Vivo Labeling, Patch Clamp, and Single Cell RNA-seq.
Correlating Anatomy and Function with Gene Expression in Individual Neurons by Combining in Vivo Labeling, Patch Clamp, and Single Cell RNA-seq.
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DOI：
10.3389/fncel.2017.00376
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发表时间：
2017
影响因子：
5.3
通讯作者：
Beltramo R
中科院分区：
文献类型：
--
作者：
Pfeffer CK;Beltramo R
将神经元分类为不同的类型是一项持续的努力，旨在揭示和了解神经系统组成部分的多样性。最近可用的方法使我们能够确定哺乳动物大脑皮层中单个神经元的基因表达模式，以生成强大的分类方案。为了彻底了解神经元的多样性，这种遗传分类方案需要与传统的分类参数相结合，如位置、轴突投影或对感觉刺激的反应特性。在这里，我们描述了一种方法，将单个神经元的基因表达与它们的位置、轴突投影或感觉反应属性联系起来。根据神经元的解剖或功能特性，在体内对神经元进行标记，并使用膜片钳移液管，分别在体外收集它们的RNA用于RNAseq。我们使用多个已建立的分子和解剖学上不同的细胞群体来验证该方法，并探索小鼠视皮层中未表征的神经元之间的分子差异。投射到额叶或对侧皮质的L5神经元之间的基因表达模式是不同的，而位置、投射或功能不同的L2神经元在分子上是相似的。利用这种方法，我们可以确定从功能和解剖上识别的单个神经元的遗传表达模式。
The classification of neurons into distinct types is an ongoing effort aimed at revealing and understanding the diversity of the components of the nervous system. Recently available methods allow us to determine the gene expression pattern of individual neurons in the mammalian cerebral cortex to generate powerful categorization schemes. For a thorough understanding of neuronal diversity such genetic categorization schemes need to be combined with traditional classification parameters like position, axonal projection or response properties to sensory stimulation. Here we describe a method to link the gene expression of individual neurons with their position, axonal projection, or sensory response properties. Neurons are labeled in vivo based on their anatomical or functional properties and, using patch clamp pipettes, their RNA individually harvested in vitro for RNAseq. We validate the methodology using multiple established molecularly and anatomically distinct cell populations and explore molecular differences between uncharacterized neurons in mouse visual cortex. Gene expression patterns between L5 neurons projecting to frontal or contralateral cortex are distinct while L2 neurons differing in position, projection, or function are molecularly similar. With this method we can determine the genetic expression pattern of functionally and anatomically identified individual neurons.
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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：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
25
作者：
Glickfeld, Lindsey L.;Andermann, Mark L.;Bonin, Vincent;Reid, R. Clay
通讯作者：
Reid, R. Clay
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