---
_id: '9438'
abstract:
- lang: eng
  text: Rigorous investigation of synaptic transmission requires analysis of unitary
    synaptic events by simultaneous recording from presynaptic terminals and postsynaptic
    target neurons. However, this has been achieved at only a limited number of model
    synapses, including the squid giant synapse and the mammalian calyx of Held. Cortical
    presynaptic terminals have been largely inaccessible to direct presynaptic recording,
    due to their small size. Here, we describe a protocol for improved subcellular
    patch-clamp recording in rat and mouse brain slices, with the synapse in a largely
    intact environment. Slice preparation takes ~2 h, recording ~3 h and post hoc
    morphological analysis 2 d. Single presynaptic hippocampal mossy fiber terminals
    are stimulated minimally invasively in the bouton-attached configuration, in which
    the cytoplasmic content remains unperturbed, or in the whole-bouton configuration,
    in which the cytoplasmic composition can be precisely controlled. Paired pre–postsynaptic
    recordings can be integrated with biocytin labeling and morphological analysis,
    allowing correlative investigation of synapse structure and function. Paired recordings
    can be obtained from mossy fiber terminals in slices from both rats and mice,
    implying applicability to genetically modified synapses. Paired recordings can
    also be performed together with axon tract stimulation or optogenetic activation,
    allowing comparison of unitary and compound synaptic events in the same target
    cell. Finally, paired recordings can be combined with spontaneous event analysis,
    permitting collection of miniature events generated at a single identified synapse.
    In conclusion, the subcellular patch-clamp techniques detailed here should facilitate
    analysis of biophysics, plasticity and circuit function of cortical synapses in
    the mammalian central nervous system.
acknowledged_ssus:
- _id: M-Shop
acknowledgement: This project received funding from the European Research Council
  (ERC) under the European Union’s Horizon 2020 research and innovation programme
  (grant agreement no. 692692 to P.J.) and the Fond zur Förderung der Wissenschaftlichen
  Forschung (Z 312-B27, Wittgenstein award to P.J., V 739-B27 to C.B.M.). We are grateful
  to F. Marr and C. Altmutter for excellent technical assistance and cell reconstruction,
  E. Kralli-Beller for manuscript editing, and the Scientific Service Units of IST
  Austria, especially T. Asenov and Miba machine shop, for maximally efficient support.
article_processing_charge: No
article_type: original
author:
- first_name: David H
  full_name: Vandael, David H
  id: 3AE48E0A-F248-11E8-B48F-1D18A9856A87
  last_name: Vandael
  orcid: 0000-0001-7577-1676
- first_name: Yuji
  full_name: Okamoto, Yuji
  id: 3337E116-F248-11E8-B48F-1D18A9856A87
  last_name: Okamoto
  orcid: 0000-0003-0408-6094
- first_name: Carolina
  full_name: Borges Merjane, Carolina
  id: 4305C450-F248-11E8-B48F-1D18A9856A87
  last_name: Borges Merjane
  orcid: 0000-0003-0005-401X
- first_name: Victor M
  full_name: Vargas Barroso, Victor M
  id: 2F55A9DE-F248-11E8-B48F-1D18A9856A87
  last_name: Vargas Barroso
- first_name: Benjamin
  full_name: Suter, Benjamin
  id: 4952F31E-F248-11E8-B48F-1D18A9856A87
  last_name: Suter
  orcid: 0000-0002-9885-6936
- first_name: Peter M
  full_name: Jonas, Peter M
  id: 353C1B58-F248-11E8-B48F-1D18A9856A87
  last_name: Jonas
  orcid: 0000-0001-5001-4804
citation:
  ama: Vandael DH, Okamoto Y, Borges Merjane C, Vargas Barroso VM, Suter B, Jonas
    PM. Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous
    pre- and postsynaptic recording at cortical synapses. <i>Nature Protocols</i>.
    2021;16(6):2947–2967. doi:<a href="https://doi.org/10.1038/s41596-021-00526-0">10.1038/s41596-021-00526-0</a>
  apa: Vandael, D. H., Okamoto, Y., Borges Merjane, C., Vargas Barroso, V. M., Suter,
    B., &#38; Jonas, P. M. (2021). Subcellular patch-clamp techniques for single-bouton
    stimulation and simultaneous pre- and postsynaptic recording at cortical synapses.
    <i>Nature Protocols</i>. Springer Nature. <a href="https://doi.org/10.1038/s41596-021-00526-0">https://doi.org/10.1038/s41596-021-00526-0</a>
  chicago: Vandael, David H, Yuji Okamoto, Carolina Borges Merjane, Victor M Vargas
    Barroso, Benjamin Suter, and Peter M Jonas. “Subcellular Patch-Clamp Techniques
    for Single-Bouton Stimulation and Simultaneous Pre- and Postsynaptic Recording
    at Cortical Synapses.” <i>Nature Protocols</i>. Springer Nature, 2021. <a href="https://doi.org/10.1038/s41596-021-00526-0">https://doi.org/10.1038/s41596-021-00526-0</a>.
  ieee: D. H. Vandael, Y. Okamoto, C. Borges Merjane, V. M. Vargas Barroso, B. Suter,
    and P. M. Jonas, “Subcellular patch-clamp techniques for single-bouton stimulation
    and simultaneous pre- and postsynaptic recording at cortical synapses,” <i>Nature
    Protocols</i>, vol. 16, no. 6. Springer Nature, pp. 2947–2967, 2021.
  ista: Vandael DH, Okamoto Y, Borges Merjane C, Vargas Barroso VM, Suter B, Jonas
    PM. 2021. Subcellular patch-clamp techniques for single-bouton stimulation and
    simultaneous pre- and postsynaptic recording at cortical synapses. Nature Protocols.
    16(6), 2947–2967.
  mla: Vandael, David H., et al. “Subcellular Patch-Clamp Techniques for Single-Bouton
    Stimulation and Simultaneous Pre- and Postsynaptic Recording at Cortical Synapses.”
    <i>Nature Protocols</i>, vol. 16, no. 6, Springer Nature, 2021, pp. 2947–2967,
    doi:<a href="https://doi.org/10.1038/s41596-021-00526-0">10.1038/s41596-021-00526-0</a>.
  short: D.H. Vandael, Y. Okamoto, C. Borges Merjane, V.M. Vargas Barroso, B. Suter,
    P.M. Jonas, Nature Protocols 16 (2021) 2947–2967.
corr_author: '1'
date_created: 2021-05-30T22:01:24Z
date_published: 2021-06-01T00:00:00Z
date_updated: 2025-04-22T22:30:43Z
day: '01'
ddc:
- '570'
department:
- _id: PeJo
doi: 10.1038/s41596-021-00526-0
ec_funded: 1
external_id:
  isi:
  - '000650528700003'
  pmid:
  - '33990799'
file:
- access_level: open_access
  checksum: 7eb580abd8893cdb0b410cf41bc8c263
  content_type: application/pdf
  creator: cziletti
  date_created: 2021-07-08T12:27:55Z
  date_updated: 2021-12-02T23:30:05Z
  embargo: 2021-12-01
  file_id: '9639'
  file_name: VandaeletalAuthorVersion2021.pdf
  file_size: 38574802
  relation: main_file
file_date_updated: 2021-12-02T23:30:05Z
has_accepted_license: '1'
intvolume: '        16'
isi: 1
issue: '6'
language:
- iso: eng
month: '06'
oa: 1
oa_version: Submitted Version
page: 2947–2967
pmid: 1
project:
- _id: 25B7EB9E-B435-11E9-9278-68D0E5697425
  call_identifier: H2020
  grant_number: '692692'
  name: Biophysics and circuit function of a giant cortical glutamatergic synapse
- _id: 25C5A090-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: Z00312
  name: Synaptic communication in neuronal microcircuits
- _id: 2696E7FE-B435-11E9-9278-68D0E5697425
  call_identifier: FWF
  grant_number: V00739
  name: Structural plasticity at mossy fiber-CA3 synapses
publication: Nature Protocols
publication_identifier:
  eissn:
  - 1750-2799
  issn:
  - 1754-2189
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous
  pre- and postsynaptic recording at cortical synapses
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 16
year: '2021'
...