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Nat Biotechnol. 2016 Feb;34(2):199-203. doi: 10.1038/nbt.3445. Epub 2015 Dec 21.

Electrophysiological, transcriptomic and morphologic profiling of single neurons using Patch-seq.

Author information

1
Department of Neuroscience, Baylor College of Medicine, Houston, Texas, USA.
2
Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.
3
Ludwig Institute for Cancer Research, Stockholm, Sweden.
4
Bernstein Center for Computational Neuroscience, Tübingen, Germany.
5
Institute for Ophthalmic Research, University of Tübingen, Tübingen, Germany.
6
Werner Reichardt Center for Integrative Neuroscience and Institute of Theoretical Physics, University of Tübingen, Tübingen, Germany.
7
Max Planck Institute for Biological Cybernetics, Tübingen, Germany.
8
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA.

Abstract

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.

PMID:
26689543
PMCID:
PMC4840019
DOI:
10.1038/nbt.3445
[Indexed for MEDLINE]
Free PMC Article
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