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

STAR Protoc. 2023 Mar 14;4(2):102168. doi: 10.1016/j.xpro.2023.102168. Online ahead of print.

Authors

Andreas Ritzau-Jost¹, Jana Nerlich², Thomas Kaas², Martin Krueger³, Timur Tsintsadze⁴, Jens Eilers², Boris Barbour⁵, Stephen M Smith⁴, Stefan Hallermann⁶

Affiliations

¹ Carl-Ludwig-Institute of Physiology, Faculty of Medicine, Leipzig University, 04103 Leipzig, Germany. Electronic address: andreas.ritzau-jost@medizin.uni-leipzig.de.
² Carl-Ludwig-Institute of Physiology, Faculty of Medicine, Leipzig University, 04103 Leipzig, Germany.
³ Institute of Anatomy, Faculty of Medicine, Leipzig University, 04103 Leipzig, Germany.
⁴ Division of Pulmonary and Critical Care Medicine, Department of Medicine, Oregon Health and Science University, Portland, OR 97239, USA; Section of Pulmonary and Critical Care Medicine, VA Portland Health Care System, Portland, OR 97239, USA.
⁵ Institut de Biologie de l'ENS (IBENS), Département de Biologie, École Normale Supérieure, CNRS, INSERM, Université PSL, 75005 Paris, France.
⁶ Carl-Ludwig-Institute of Physiology, Faculty of Medicine, Leipzig University, 04103 Leipzig, Germany. Electronic address: hallermann@medizin.uni-leipzig.de.

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.

Keywords: Biophysics; Cell Biology; Cell Culture; Neuroscience.

Abstract

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