--- res: bibo_abstract: - 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.@eng bibo_authorlist: - foaf_Person: foaf_givenName: David H foaf_name: Vandael, David H foaf_surname: Vandael foaf_workInfoHomepage: http://www.librecat.org/personId=3AE48E0A-F248-11E8-B48F-1D18A9856A87 orcid: 0000-0001-7577-1676 - foaf_Person: foaf_givenName: Yuji foaf_name: Okamoto, Yuji foaf_surname: Okamoto foaf_workInfoHomepage: http://www.librecat.org/personId=3337E116-F248-11E8-B48F-1D18A9856A87 orcid: 0000-0003-0408-6094 - foaf_Person: foaf_givenName: Carolina foaf_name: Borges Merjane, Carolina foaf_surname: Borges Merjane foaf_workInfoHomepage: http://www.librecat.org/personId=4305C450-F248-11E8-B48F-1D18A9856A87 orcid: 0000-0003-0005-401X - foaf_Person: foaf_givenName: Victor M foaf_name: Vargas Barroso, Victor M foaf_surname: Vargas Barroso foaf_workInfoHomepage: http://www.librecat.org/personId=2F55A9DE-F248-11E8-B48F-1D18A9856A87 - foaf_Person: foaf_givenName: Benjamin foaf_name: Suter, Benjamin foaf_surname: Suter foaf_workInfoHomepage: http://www.librecat.org/personId=4952F31E-F248-11E8-B48F-1D18A9856A87 orcid: 0000-0002-9885-6936 - foaf_Person: foaf_givenName: Peter M foaf_name: Jonas, Peter M foaf_surname: Jonas foaf_workInfoHomepage: http://www.librecat.org/personId=353C1B58-F248-11E8-B48F-1D18A9856A87 orcid: 0000-0001-5001-4804 bibo_doi: 10.1038/s41596-021-00526-0 bibo_issue: '6' bibo_volume: 16 dct_date: 2021^xs_gYear dct_identifier: - UT:000650528700003 dct_isPartOf: - http://id.crossref.org/issn/17542189 - http://id.crossref.org/issn/17502799 dct_language: eng dct_publisher: Springer Nature@ dct_title: Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous pre- and postsynaptic recording at cortical synapses@ fabio_hasPubmedId: '33990799' ...