Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures

Andreas Ritzau-Jost; Jana Nerlich; Thomas Kaas; Martin Krueger; Timur Tsintsadze; Jens Eilers; Boris Barbour; Stephen M. Smith; Stefan Hallermann

doi:10.1016/j.xpro.2023.102168

Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures

Andreas Ritzau-Jost
, Jana Nerlich
, Thomas Kaas
, Martin Krueger
, Timur Tsintsadze
, Jens Eilers
, Boris Barbour
, Stephen M. Smith
, Stefan Hallermann

Pulmonary and Critical Care Medicine

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details on precise pipette capacitance compensation required for high-resolution current-clamp recordings from boutons. For further details on the use and execution of this protocol, please refer to Ritzau-Jost et al.¹

Original language	English (US)
Article number	102168
Journal	STAR Protocols
Volume	4
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2023.102168
State	Published - Jun 16 2023

Funding

This work was supported by a European Research Council Consolidator Grant ( ERC CoG 865634 ) to S.H., the German Research Foundation ( HA6386/10-1 to S.H.), and by grants awarded by the U.S. Department of Veterans Affairs ( BX002547 ) and NIGMS ( GM134110 ) to S.M.S.

Funders	Funder number
National Institute of General Medical Sciences	GM134110
U.S. Department of Veterans Affairs	BX002547
H2020 European Research Council	865634
Deutsche Forschungsgemeinschaft	HA6386/10-1

Keywords

Biophysics
Cell Biology
Cell Culture
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology

Access to Document

10.1016/j.xpro.2023.102168

Cite this

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title = "Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures",

abstract = "Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details on precise pipette capacitance compensation required for high-resolution current-clamp recordings from boutons. For further details on the use and execution of this protocol, please refer to Ritzau-Jost et al.1",

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AU - Ritzau-Jost, Andreas

AU - Nerlich, Jana

AU - Kaas, Thomas

AU - Krueger, Martin

AU - Tsintsadze, Timur

AU - Eilers, Jens

AU - Barbour, Boris

AU - Smith, Stephen M.

AU - Hallermann, Stefan

PY - 2023/6/16

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N2 - Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details on precise pipette capacitance compensation required for high-resolution current-clamp recordings from boutons. For further details on the use and execution of this protocol, please refer to Ritzau-Jost et al.1

AB - Direct electrical recordings from conventional boutons in the mammalian central nervous system have proven challenging due to their small size. Here, we provide a protocol for direct whole-cell patch-clamp recordings from small presynaptic boutons of primary dissociated cultured neurons of the rodent neocortex. We describe steps to prepare primary neocortical cultures and recording pipettes, followed by identifying boutons and establishing a whole-cell bouton recording. We then provide details on precise pipette capacitance compensation required for high-resolution current-clamp recordings from boutons. For further details on the use and execution of this protocol, please refer to Ritzau-Jost et al.1

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KW - Cell Biology

KW - Cell Culture

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Direct whole-cell patch-clamp recordings from small boutons in rodent primary neocortical neuron cultures

Abstract

Funding

Keywords

ASJC Scopus subject areas

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