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

• Published in: STAR protocols Vol. 4; no. 2; p. 102168
• Main Authors: Ritzau-Jost, Andreas, Nerlich, Jana, Kaas, Thomas, Krueger, Martin, Tsintsadze, Timur, Eilers, Jens, Barbour, Boris, Smith, Stephen M., Hallermann, Stefan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16-06-2023; Elsevier
• Subjects: Biophysics; Cell Biology; Cell Culture; Life Sciences; Neurons and Cognition; Neuroscience; Protocol; Cell Culture; Neuroscience; Biophysics; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102168

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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•Identification and whole-cell recording from small boutons in mouse neuronal cultures•Optimization and compensation of pipette capacitance for high-resolution recordings•Adaptations of recording equipment for small bouton recordings•Applicable to small boutons of cultures and acute brain slices of other brain areas
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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.