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Related Experiment Video

Updated: May 30, 2025

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution
08:48

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An improved FLARE system for recording and manipulating neuronal activity.

Guanwei Zhou1,2,3, Ruonan Li1,2,3, Ola Bartolik1,4

  • 1Life Sciences Institute, University of Michigan, Ann Arbor, MI, USA.

Biorxiv : the Preprint Server for Biology
|January 27, 2025
PubMed
Summary

Researchers developed cytoFLARE, a novel tool for precisely labeling and controlling neuronal activity in real-time. This improved system enhances the study of neural circuits underlying cognition and behavior in vivo.

Keywords:
DrosophilaIntegrator sensorNeuronal activityTranscriptional reporter

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • * Recording and manipulating neuronal ensembles in vivo is crucial for understanding cognition and behavior.
  • * Existing tools like FLARE (Fluorescently Labeled Activated Reporter) have limitations in sensitivity to prolonged neuronal activity.

Purpose of the Study:

  • * To develop an improved version of FLARE, named cytoFLARE, for enhanced neuronal activity reporting.
  • * To characterize cytoFLARE's performance in cellular and in vivo models.
  • * To demonstrate the application of cytoFLARE for precise temporal control of neuronal ensembles.

Main Methods:

  • * Engineering cytoFLARE with cytosolic transcription factor expression and enhanced calcium-sensing domains.
  • * Validating cytoFLARE in HEK293T cells and primary neuronal cultures.
  • * Establishing cytoFLARE transgenic Drosophila models.
  • * Applying cytoFLARE for time-gated neuronal labeling upon sensory or optogenetic stimulation.
  • * Utilizing cytoFLARE to drive optogenetic actuator expression for neuronal reactivation.

Main Results:

  • * cytoFLARE demonstrated improved calcium- and light-dependent signaling in HEK293T cells.
  • * Higher signal-to-background ratios were observed in neuronal cultures compared to FLARE.
  • * Successful time-gated labeling of activated neurons in Drosophila models.
  • * Demonstrated reactivation of specific neurons in the larval nociceptive system using cytoFLARE.

Conclusions:

  • * cytoFLARE represents a significant advancement over FLARE, offering enhanced sensitivity and precision.
  • * This study presents the first characterization and application of time-gated calcium integrators in Drosophila.
  • * cytoFLARE provides a powerful new tool for dissecting neural circuits and their functions.