Cited by 60Neuroscience and Neuropharmacology ResearchCellular transport and secretionAdvanced Electron Microscopy Techniques and Applications

Functional Electron Microscopy, “Flash and Freeze,” of Identified Cortical Synapses in Acute Brain Slices

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Borges-Merjane, Carolina, Kim, Olena, Jonas, Peter·2020·Neuron

Abstract

How structural and functional properties of synapses relate to each other is a fundamental question in neuroscience. Electrophysiology has elucidated mechanisms of synaptic transmission, and electron microscopy (EM) has provided insight into morphological properties of synapses. Here we describe an enhanced method for functional EM ("flash and freeze"), combining optogenetic stimulation with high-pressure freezing. We demonstrate that the improved method can be applied to intact networks in acute brain slices and organotypic slice cultures from mice. As a proof of concept, we probed vesicle pool changes during synaptic transmission at the hippocampal mossy fiber-CA3 pyramidal neuron synapse. Our findings show overlap of the docked vesicle pool and the functionally defined readily releasable pool and provide evidence of fast endocytosis at this synapse. Functional EM with acute slices and slice cultures has the potential to reveal the structural and functional mechanisms of transmission in intact, genetically perturbed, and disease-affected synapses.

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Authors
Borges-Merjane, C., Kim, O., & Jonas, P.
Affiliations
Institute of Science and Technology Austria (AT)
Corresponding author
Borges-Merjane, Carolina
Venue
Neuron
Publisher
Elsevier BV
Published
2020-03
Volume
105
Issue
6
Pages
992-1006.e6
ISSN
0896-6273 (ISSN-L)
OpenAlex
W2998834324
Cited by
60
References
61
Citations by year
2026: 6 · 2025: 8 · 2024: 11 · 2023: 11 · 2022: 9 · 2021: 10 · 2020: 5
FWCI
4.01
Type
journal article
Language
en
Update notices
erratum (2020-12-09)
Open access
Locations
  • Neuron (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Horizon 2020
  • European Research Council (692692)
  • H2020 Marie Skłodowska-Curie Actions (708497)
  • Austrian Science Fund (DK W 1205-B09, Z 312-B27)
  • Horizon 2020 Framework Programme
  • European Commission
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
Keywords
optogenetic stimulation, high-pressure freezing, synaptic transmission, readily releasable pool, mossy fiber-CA3 synapses, hippocampus, acute brain slices, organotypic slice cultures
MeSH terms
Animals, Cerebral Cortex, Endocytosis, Freeze Fracturing, Microscopy, Electron, Synapses, Synaptic Vesicles, Pyramidal Cells, Mossy Fibers, Hippocampal, Mice, Functional Neuroimaging, Optogenetics

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