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Related Experiment Video

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Multimodal imaging of synaptic vesicles with a single probe.

Seong J An1,2, Massimiliano Stagi3,4,5,6, Travis J Gould2,7

  • 1Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06510, USA.

Cell Reports Methods
|May 2, 2022
PubMed
Summary

Researchers developed a versatile synaptic vesicle probe using a C2 domain. This single, modular probe allows imaging across multiple microscopy techniques, aiding the study of synaptic vesicle recycling and membrane dynamics.

Keywords:
C2 domainPALMSTEDTIRFelectron microscopysuper-resolutionsynapsesynaptic vesicles

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Understanding synaptic-vesicle recycling is crucial for neuroscience.
  • Current microscopy probes often target specific modalities, limiting comprehensive analysis.
  • A need exists for versatile probes to study vesicle dynamics across different resolutions.

Purpose of the Study:

  • To develop a single, modular probe for imaging synaptic vesicles using multiple microscopy techniques.
  • To overcome limitations of modality-specific probes in studying vesicle recycling.
  • To provide a tool for high-resolution imaging of synaptic vesicle function.

Main Methods:

  • Utilized the membrane-binding C2 domain of cytosolic phospholipase A2 (cPLA2).
  • Conjugated the C2 domain with various detectable tags.
  • Demonstrated imaging of synaptic vesicles with the modular probe.

Main Results:

  • A single, modular probe based on the cPLA2 C2 domain was successfully created.
  • The probe enabled synaptic vesicle imaging across multiple spatial and temporal resolutions.
  • The C2 domain functions as a general endocytic marker.

Conclusions:

  • The developed cPLA2 C2 domain probe offers a versatile tool for synaptic vesicle research.
  • This modular probe complements existing tools for studying vesicle recycling and membrane dynamics.
  • The C2 domain's utility extends to studying membrane recycling in diverse cell types.