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Resolving synaptic events using subsynaptically targeted GCaMP8 variants.

Jiawen Chen1,2, Junhao Lin1, Kaikai He1,2

  • 1University of Southern California, Department of Neurobiology, Los Angeles, United States.

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|March 2, 2026
PubMed
Summary
This summary is machine-generated.

Next-generation genetically encoded calcium indicators (GCaMP8) and the CaFire analysis tool significantly improve the speed and sensitivity of visualizing neural activity at synaptic compartments in Drosophila.

Keywords:
D. melanogastercalciumgenetically-encodedindicatorneuromuscular junctionneuroscienceratiometricsynaptic terminals

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Genetically encoded calcium indicators (GCaMP) are crucial for visualizing neural activity.
  • Previous GCaMP versions and synthetic dyes had limitations in speed and sensitivity, especially at synaptic compartments.
  • Electrophysiology is a gold standard but lacks spatial resolution.

Purpose of the Study:

  • To engineer and validate next-generation GCaMP8-based indicators for synaptic compartments.
  • To develop an automated analysis program (CaFire) for quantifying calcium signals.
  • To demonstrate the improved performance of GCaMP8 sensors compared to existing methods.

Main Methods:

  • Engineering GCaMP8 indicators targeted to presynaptic boutons, active zones, and postsynaptic compartments in Drosophila.
  • Validation of sensor performance against previous GCaMP versions and synthetic dyes.
  • Development of the Python-based CaFire program for automated signal quantification.
  • Application of CaFire to analyze presynaptic, active zone, and postsynaptic calcium dynamics.

Main Results:

  • Engineered GCaMP8 sensors demonstrated superior performance compared to previous versions and synthetic dyes.
  • The CaFire program enabled automated quantification of evoked and spontaneous calcium signals.
  • A presynaptic GCaMP8m sensor accurately captured physiologically relevant calcium changes with high sensitivity.
  • An active zone-targeted sensor distinguished calcium differences between release sites.
  • A postsynaptic GCaMP8m sensor detected quantal events with resolution comparable to electrophysiology.

Conclusions:

  • Next-generation GCaMP8 sensors, when targeted to synaptic compartments, achieve speed and sensitivity rivaling chemical dyes and electrophysiology.
  • The CaFire analysis tool facilitates robust quantification of synaptic calcium dynamics.
  • These advancements enable high-resolution investigation of calcium signaling at the synapse.