Cited by 66Neuroscience and Neuropharmacology ResearchCellular transport and secretionAdvanced Electron Microscopy Techniques and Applications

Ultrastructural Imaging of Activity-Dependent Synaptic Membrane-Trafficking Events in Cultured Brain Slices

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Imig, Cordelia, López-Murcia, Francisco José, Maus, Lydia, García-Plaza, Inés Hojas, Mortensen, Lena Sünke, Schwark, Manuela, Schwarze, Valentin, Angibaud, Julie +4 more·2020·Neuron

Abstract

Electron microscopy can resolve synapse ultrastructure with nanometer precision, but the capture of time-resolved, activity-dependent synaptic membrane-trafficking events has remained challenging, particularly in functionally distinct synapses in a tissue context. We present a method that combines optogenetic stimulation-coupled cryofixation ("flash-and-freeze") and electron microscopy to visualize membrane trafficking events and synapse-state-specific changes in presynaptic vesicle organization with high spatiotemporal resolution in synapses of cultured mouse brain tissue. With our experimental workflow, electrophysiological and "flash-and-freeze" electron microscopy experiments can be performed under identical conditions in artificial cerebrospinal fluid alone, without the addition of external cryoprotectants, which are otherwise needed to allow adequate tissue preservation upon freezing. Using this approach, we reveal depletion of docked vesicles and resolve compensatory membrane recycling events at individual presynaptic active zones at hippocampal mossy fiber synapses upon sustained stimulation.

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Authors
Imig, C., López-Murcia, F. J., Maus, L., García-Plaza, I. H., Mortensen, L. S., Schwark, M., Schwarze, V., Angibaud, J., Nägerl, U. V., Taschenberger, H., Brose, N., & Cooper, B. H.
Affiliations
  • Max Planck Institute of Experimental Medicine (DE)
  • Centre National de la Recherche Scientifique (FR)
Corresponding author
  • Imig, Cordelia
  • Brose, Nils
  • Cooper, Benjamin H.
Venue
Neuron
Publisher
Elsevier BV
Published
2020-12
Volume
108
Issue
5
Pages
843-860.e8
ISSN
0896-6273 (ISSN-L)
OpenAlex
W3088769972
Cited by
66
References
85
Citations by year
2026: 6 · 2025: 11 · 2024: 10 · 2023: 13 · 2022: 14 · 2021: 11 · 2020: 1
FWCI
4.47
Type
journal article
Language
en
Open access
Locations
  • Neuron (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
  • Dipòsit Digital de la Universitat de Barcelona (Universitat de Barcelona) (submittedVersion, cc-by-nc-nd)
  • GoeScholar The Publication Server of the Georg-August-Universität Göttingen (Georg-August-Universität Göttingen) (submittedVersion, cc-by)
  • MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Funders
  • European Commission (ERC Advanced Grant SynPrime)
  • Deutsche Forschungsgemeinschaft (SFB1286/A01)
  • Germany’s Excellence Strategy (EXC 2067/1-390729940)
  • IMPRS Neurosciences
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
presynaptic vesicles, optogenetic stimulation, cryofixation, active zone, docked vesicles, membrane recycling, synaptic ultrastructure
MeSH terms
Animals, Hippocampus, Mice, Transgenic, Microscopy, Electron, Microtomy, Organ Culture Techniques, Synaptic Membranes, Excitatory Postsynaptic Potentials, Protein Transport, Mice, Gene Knock-In Techniques

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