Presynaptic cAMP-PKA-mediated potentiation induces reconfiguration of synaptic vesicle pools and channel-vesicle coupling at hippocampal mossy fiber boutons

PLoS Biol. 2024 Nov 18;22(11):e3002879. doi: 10.1371/journal.pbio.3002879. eCollection 2024 Nov.

Abstract

It is widely believed that information storage in neuronal circuits involves nanoscopic structural changes at synapses, resulting in the formation of synaptic engrams. However, direct evidence for this hypothesis is lacking. To test this conjecture, we combined chemical potentiation, functional analysis by paired pre-postsynaptic recordings, and structural analysis by electron microscopy (EM) and freeze-fracture replica labeling (FRL) at the rodent hippocampal mossy fiber synapse, a key synapse in the trisynaptic circuit of the hippocampus. Biophysical analysis of synaptic transmission revealed that forskolin-induced chemical potentiation increased the readily releasable vesicle pool size and vesicular release probability by 146% and 49%, respectively. Structural analysis of mossy fiber synapses by EM and FRL demonstrated an increase in the number of vesicles close to the plasma membrane and the number of clusters of the priming protein Munc13-1, indicating an increase in the number of both docked and primed vesicles. Furthermore, FRL analysis revealed a significant reduction of the distance between Munc13-1 and CaV2.1 Ca2+ channels, suggesting reconfiguration of the channel-vesicle coupling nanotopography. Our results indicate that presynaptic plasticity is associated with structural reorganization of active zones. We propose that changes in potential nanoscopic organization at synaptic vesicle release sites may be correlates of learning and memory at a plastic central synapse.

MeSH terms

  • Animals
  • Calcium Channels, N-Type / metabolism
  • Colforsin / pharmacology
  • Cyclic AMP* / metabolism
  • Cyclic AMP-Dependent Protein Kinases* / metabolism
  • Hippocampus / cytology
  • Hippocampus / metabolism
  • Hippocampus / physiology
  • Male
  • Mice
  • Mossy Fibers, Hippocampal* / metabolism
  • Mossy Fibers, Hippocampal* / physiology
  • Mossy Fibers, Hippocampal* / ultrastructure
  • Nerve Tissue Proteins / metabolism
  • Neuronal Plasticity / physiology
  • Presynaptic Terminals* / metabolism
  • Presynaptic Terminals* / physiology
  • Rats
  • Synaptic Transmission / physiology
  • Synaptic Vesicles* / metabolism
  • Synaptic Vesicles* / physiology
  • Synaptic Vesicles* / ultrastructure

Substances

  • Cyclic AMP-Dependent Protein Kinases
  • Cyclic AMP
  • Nerve Tissue Proteins
  • Calcium Channels, N-Type
  • Colforsin
  • Unc13a protein, mouse

Grants and funding

This project received funding from the European Research Council and European Union’s Horizon 2020 research and innovation programme (ERC 692692 to P.J.; https://cordis.europa.eu/project/id/692692/de) and from the Fond zur Förderung der Wissenschaftlichen Forschung (Z312-B27 Wittgenstein award to P.J., https://www.fwf.ac.at/en/funding/portfolio/projects/fwf-wittgenstein-award; W1205-B09 and P36232-B to P.J., https://www.fwf.ac.at/en/funding; I6166-B to R.S.; https://www.fwf.ac.at/en/funding). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.