Patch-Seq Protocol to Analyze the Electrophysiology, Morphology and Transcriptome of Whole Single Neurons Derived From Human Pluripotent Stem Cells

Frontiers in Molecular Neuroscience
M van den HurkC Bardy

Abstract

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 huma...Continue Reading

References

Aug 1, 1992·Neuron·B LambolezJ Rossier
Apr 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·J EberwineP Coleman
Jan 11, 2001·Cell and Tissue Research·N J SucherE Guenther
Sep 5, 2002·The Journal of Comparative Neurology·Bassam N HamamAngel A Alonso
Apr 30, 2005·Nature Reviews. Neuroscience·Claudia Bagni, William T Greenough
Dec 17, 2008·Proceedings of the National Academy of Sciences of the United States of America·Nicholas M TorielloRichard A Mathies
May 14, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ju-Yun WengCheng-Chang Lien
Feb 1, 2011·Proceedings of the National Academy of Sciences of the United States of America·Ji-Eun KimAnirvan Ghosh
Apr 29, 2011·Proceedings of the National Academy of Sciences of the United States of America·Wenlin LiSheng Ding
Sep 6, 2011·American Journal of Physiology. Heart and Circulatory Physiology·Junyi MaCraig T January
Sep 10, 2011·Expert Review of Molecular Diagnostics·Anders StåhlbergPierre Aman
Dec 24, 2011·Nature Protocols·Ami CitriRobert C Malenka
Aug 9, 2012·Current Protocols in Stem Cell Biology·Leah F BoyerFred H Gage
Jan 29, 2013·Frontiers in Neural Circuits·Melanie GingerAndreas Frick
Feb 14, 2014·Nature Methods·James EberwineJunhyong Kim
Jun 14, 2014·Science·Anoop P PatelBradley E Bernstein
Aug 29, 2014·PloS One·Christina R MuratoreTracy L Young-Pearse
Oct 7, 2014·Journal of Visualized Experiments : JoVE·Pengbo ZhangRenee A Reijo Pera
Apr 15, 2015·Proceedings of the National Academy of Sciences of the United States of America·Cedric BardyFred H Gage
Oct 28, 2015·Molecular Psychiatry·K C VadodariaF H Gage
Dec 23, 2015·Nature Biotechnology·Cathryn R CadwellAndreas S Tolias
Aug 18, 2016·Proceedings of the National Academy of Sciences of the United States of America·Csaba FöldyThomas C Südhof

❮ Previous
Next ❯

Citations

Jun 4, 2020·Molecular Autism·Danijela DrakulicAdrian J Harwood
Jan 17, 2020·International Journal of Molecular Sciences·Bruno SterliniAnna Corradi
Oct 24, 2018·Frontiers in Molecular Neuroscience·Shreejoy J TripathyPaul Pavlidis
Dec 31, 2020·STAR Protocols·Marcela LipovsekMatthew S Grubb
Sep 25, 2020·Neuron·Ashley Del DossoGiorgia Quadrato
Feb 9, 2021·IScience·Jonghun KimIn-Hyun Park
Mar 2, 2019·Seminars in Cell & Developmental Biology·Yang D Teng
May 6, 2021·Journal of Personalized Medicine·Wanda LattanziOrnella Parolini
Oct 14, 2021·Nature Communications·Zhongxing SunDong Fang
Oct 22, 2020··Matthew S. GrubbMarcela Lipovsek

❮ Previous
Next ❯

Methods Mentioned

BETA
reverse transcription PCR
scRNA-seq
Patch-seq
biopsy
RNA-seq
PCR
Illumina Sequencing
Smart-seq
Assay
Chip

Software Mentioned

SMARTer
SMART
Neurolucida
Patch
seq
Qubit

Related Concepts

Related Feeds

CZI Human Cell Atlas Seed Network

The aim of the Human Cell Atlas (HCA) is to build reference maps of all human cells in order to enhance our understanding of health and disease. The Seed Networks for the HCA project aims to bring together collaborators with different areas of expertise in order to facilitate the development of the HCA. Find the latest research from members of the HCA Seed Networks here.