High-yield in vitro recordings from neurons functionally characterized in vivo

Nature Protocols
Simon WeilerVolker Scheuss

Abstract

In vivo two-photon calcium imaging provides detailed information about the activity and response properties of individual neurons. However, in vitro methods are often required to study the underlying neuronal connectivity and physiology at the cellular and synaptic levels at high resolution. This protocol provides a fast and reliable workflow for combining the two approaches by characterizing the response properties of individual neurons in mice in vivo using genetically encoded calcium indicators (GECIs), followed by retrieval of the same neurons in brain slices for further analysis in vitro (e.g., circuit mapping). In this approach, a reference frame is provided by fluorescent-bead tracks and sparsely transduced neurons expressing a structural marker in order to re-identify the same neurons. The use of GECIs provides a substantial advancement over previous approaches by allowing for repeated in vivo imaging. This opens the possibility of directly correlating experience-dependent changes in neuronal activity and feature selectivity with changes in neuronal connectivity and physiology. This protocol requires expertise both in in vivo two-photon calcium imaging and in vitro electrophysiology. It takes 3 weeks or more to complete...Continue Reading

References

Apr 6, 1990·Science·W DenkW W Webb
Jan 1, 1997·Spatial Vision·D H Brainard
Jan 1, 1997·Methods in Molecular Biology·T Wang, I S Kass
Jun 5, 2003·Proceedings of the National Academy of Sciences of the United States of America·Christoph StosiekArthur Konnerth
Jun 13, 2003·Biomedical Engineering Online·Thomas A PologrutoKarel Svoboda
Feb 12, 2004·Science's STKE : Signal Transduction Knowledge Environment·Ryohei YasudaKarel Svoboda
Jul 6, 2004·Journal of Virological Methods·Refael ItahClaytus Davis
Jul 20, 2005·PLoS Biology·Carlos Portera-CailliauKarel Svoboda
Dec 8, 2005·Visual Neuroscience·Jianhua CangMichael P Stryker
Jun 20, 2006·Journal of Neuroscience Methods·James G Canfield
Jul 25, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Cristopher M Niell, Michael P Stryker
Sep 6, 2008·Science·Eva PastalkovaGyörgy Buzsáki
Jan 20, 2009·Nature·Leopoldo PetreanuKarel Svoboda
Jan 12, 2011·Neuroinformatics·Hermann CuntzMichael Häusser
Jul 12, 2011·Nature Neuroscience·Wolfgang MittmannJason N D Kerr
Jul 19, 2011·Nature Neuroscience·Sonja B HoferThomas D Mrsic-Flogel
Jan 1, 2010·Frontiers in Neural Circuits·Benjamin A SuterGordon M G Shepherd
Dec 3, 2011·Pflügers Archiv : European journal of physiology·Dominik Langer, Fritjof Helmchen
Dec 7, 2011·Frontiers in Neural Circuits·Anthony D Lien, Massimo Scanziani
Mar 13, 2012·Neuron·Christine Grienberger, Arthur Konnerth
Nov 12, 1986·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·J G WhiteS Brenner
Sep 6, 2012·Cold Spring Harbor Protocols·Gordon M G Shepherd
Oct 5, 2012·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Jasper AkerboomLoren L Looger
Feb 12, 2013·Nature Neuroscience·Yaniv ZivMark J Schnitzer
Apr 5, 2013·Nature·Ho KoThomas D Mrsic-Flogel
Jun 4, 2013·Nature Neuroscience·J Tiago GonçalvesCarlos Portera-Cailliau
Jul 23, 2013·Nature·Tsai-Wen ChenDouglas S Kim
Aug 27, 2013·Nature·Sandra J KuhlmanJoshua T Trachtenberg
Mar 19, 2014·Journal of Visualized Experiments : JoVE·Marcus LeinweberGeorg B Keller
May 20, 2014·Nature Methods·Robert PrevedelAlipasha Vaziri
Sep 25, 2014·PloS One·Hod DanaKarel Svoboda
Oct 3, 2014·Nature Protocols·Glenn J GoldeyMark L Andermann
Dec 6, 2014·Frontiers in Molecular Neuroscience·Tobias RoseOliver Griesbeck
Jan 20, 2015·Nature Neuroscience·Gordon B SmithDavid Fitzpatrick

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Citations

Feb 26, 2019·Nature Materials·Xiao YangCharles M Lieber
Jun 4, 2019·Frontiers in Synaptic Neuroscience·Jean-Sébastien Jouhanneau, James F A Poulet
Jun 11, 2019·Frontiers in Synaptic Neuroscience·Christian EbnerRobert N S Sachdev
May 15, 2020·Journal of Neuroscience Methods·Philip AnnerGeorg Dorffner
Jul 30, 2021·Current Biology : CB·Annet GlasPieter M Goltstein

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