Combined two-photon imaging, electrophysiological, and anatomical investigation of the human neocortex in vitro
Open accessAbstract
Spontaneous synchronous population activity (SPA) can be detected by electrophysiological methods in cortical slices of epileptic patients, maintained in a physiological medium in vitro. In order to gain additional spatial information about the network mechanisms involved in the SPA generation, we combined electrophysiological studies with two-photon imaging. Neocortical slices prepared from postoperative tissue of epileptic and tumor patients were maintained in a dual perfusion chamber in a physiological incubation medium. SPA was recorded with a 24-channel extracellular linear microelectrode covering all neocortical layers. After identifying the electrophysiologically active regions of the slice, bolus loading of neuronal and glial markers was applied on the tissue. SPA-related Ca2+ transients were detected in a large population of neighboring neurons with two-photon microscopy, simultaneous with extracellular SPA and intracellular whole-cell patch-clamp recordings. The intracellularly recorded cells were filled for subsequent anatomy. The cells were reconstructed in three dimensions and examined with light- and transmission electron microscopy. Combining high spatial resolution two-photon Ca2+ imaging techniques and high temporal resolution extra- and intracellular electrophysiology with cellular anatomy may permit a deeper understanding of the structural and functional properties of the human neocortex.
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Backpropagation of action potentials to dendrites of human neurons was confirmed by detecting dendritic Ca2+ responses following somatically elicited burst firing in layer 3 pyramidal cells.
“Backpropagation of action potentials to dendrites of human neurons has been shown in previous studies13,29 and we confirmed these results by detecting dendritic Ca2+ responses following somatically elicited burst firing (100 ms current injections, 4 spikes/burst) in layer 3 pyramidal cells.”
From Transcriptomic and morphophysiological evidence for a specialized human cortical GABAergic cell type (2018) · cites this paper as [29]
Human pyramidal cells have uniquely small membrane capacitance, which promotes backpropagation of action potentials and increases excitability in human dendrites relative to rodent dendrites with larger capacitance.
“The uniquely small membrane capacitance (Cm) found in human pyramidal cells8 promotes backpropagation of action potentials and increases excitability in human dendrites13,29 relative to rodent dendrites having larger Cm.”
From Transcriptomic and morphophysiological evidence for a specialized human cortical GABAergic cell type (2018) · cites this paper as [29]
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- Authors
- Kerekes, B. P., Toth, K., Kaszás, A., Chiovini, B., Szadai, Z., Szalay, G., Palfi, D., Spitzer, K., Rózsa, B., Ulbert, I., & Wittner, L.
- Affiliations
- Pázmány Péter Catholic University (HU)
- Hungarian Academy of Sciences
- HUN-REN Institute of Experimental Medicine (HU)
- Pázmány Péter Catholic Univ.
- Országos Idegsebészeti Tudományos Intézet (HU)
- Venue
- Neurophotonics
- Publisher
- SPIE-Intl Soc Optical Eng
- Published
- 2014-09-11
- Volume
- 1
- Issue
- 01
- Pages
- 1-10
- ISSN
- 2329-423X (ISSN-L)
- 2329-4248
- PMC
- PMC4478968
- OpenAlex
- W2101608461
- Cited by
- 16
- References
- 55
- Citations by year
- 2026: 1 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 2 · 2020: 1 · 2019: 4 · 2018: 1 · 2017: 1 · 2016: 1 · 2015: 1
- FWCI
- 0.56
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Neurophotonics (publishedVersion, cc-by)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Hungarian-French (TÉT_10-1-2011-0389, GOP-1.1.1-08/1-2008-0085)
- Swiss-Hungarian (SH/7/2/8, KTIA (KMR_12-1-2012-0214))
- Hungarian (OTKA PD91151)
- Bolyai Research Fellowship (KTIA_NAP_13, TÁMOP 4.2.1.B11/2/KMR2011-0002, TÁMOP 4.2.4.A/1-11-1-2012-0001)
- Nemzeti Kiválóság Program (FP7-ICT-2011-C 323945)
- Topics
- Life Sciences › Neuroscience › Cognitive Neuroscience › Neural dynamics and brain function
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Health Sciences › Medicine › Psychiatry and Mental health › Epilepsy research and treatment
- Keywords
- human neocortex, two-photon calcium imaging, whole-cell patch clamp, three-dimensional cell reconstruction, epileptogenesis
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