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Updated: May 28, 2026
Vibrodissociation of Neurons from Rodent Brain Slices to Study Synaptic Transmission and Image Presynaptic Terminals
Published on: May 25, 2011
Characterization of MSB synapses in dissociated hippocampal culture with simultaneous pre- and postsynaptic live
James E Reilly1, Hugo H Hanson, Mónica Fernández-Monreal
1Department of Neuroscience, Mount Sinai School of Medicine, New York City, New York, United States of America.
Abstract:
Multisynaptic boutons (MSBs) are presynaptic boutons in contact with multiple postsynaptic partners. Although MSB synapses have been studied with static imaging techniques such as electron microscopy (EM), the dynamics of individual MSB synapses have not been directly evaluated. It is known that the number of MSB synapses increases with synaptogenesis and plasticity but the formation, behavior, and fate of individual MSB synapses remains largely unknown. To address this, we developed a means of live imaging MSB synapses to observe them directly over time. With time lapse confocal microscopy of GFP-filled dendrites in contact with VAMP2-DsRed-labeled boutons, we recorded both MSBs and their contacting spines hourly over 15 or more hours. Our live microscopy showed that, compared to spines contacting single synaptic boutons (SSBs), MSB-contacting spines exhibit elevated dynamic behavior. These results are consistent with the idea that MSBs serve as intermediates in synaptic development and plasticity.
Insights
Multisynaptic boutons (MSBs) show increased dynamic behavior in their contacting spines compared to single synaptic boutons (SSBs). This live imaging study reveals MSBs as key players in synaptic development and plasticity.
Area of Science:
- Neuroscience
- Cell Biology
- Synaptic Plasticity
Background:
- Multisynaptic boutons (MSBs) are presynaptic structures contacting multiple postsynaptic partners.
- Previous studies on MSBs relied on static imaging, limiting understanding of their dynamic nature.
- The formation, behavior, and fate of individual MSB synapses remain largely unknown.
Purpose of the Study:
- To develop and utilize a live imaging technique to observe the dynamics of individual MSB synapses.
- To compare the dynamic behavior of spines contacting MSBs versus single synaptic boutons (SSBs).
Main Methods:
- Developed a live imaging approach using time-lapse confocal microscopy.
- Visualized GFP-filled dendrites and VAMP2-DsRed-labeled boutons.
- Recorded MSBs and their contacting spines hourly for over 15 hours.
Main Results:
- MSB-contacting spines exhibited significantly elevated dynamic behavior compared to spines contacting SSBs.
- Live imaging provided direct temporal evaluation of individual MSB synapse dynamics.
- Observed hourly dynamics over extended periods revealed novel insights into synapse behavior.
Conclusions:
- MSBs play a crucial role in synaptic development and plasticity.
- The dynamic nature of MSB-contacting spines suggests an active role in synaptic remodeling.
- Live imaging is a powerful tool for studying synapse dynamics and function.
