Functional Electron Microscopy, “Flash and Freeze,” of Identified Cortical Synapses in Acute Brain Slices
Open accessAbstract
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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The mossy fiber-CA3 PN synapse is accessible to quantitative structural analysis by electron microscopy, high-pressure freezing, and freeze-fracture replica labeling.
“Finally, it is accessible to quantitative structural analysis by EM [34,35], high-pressure freezing (HPF) [36–38], and freeze-fracture replica labeling (FRL) [39].”
From Presynaptic cAMP-PKA-mediated potentiation induces reconfiguration of synaptic vesicle pools and channel-vesicle coupling at hippocampal mossy fiber boutons (2024) · cites this paper as [37]
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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)
- PMC
- PMC7083231
- 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
- hybrid
- doi.org
- Licence
- 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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