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Updated: Dec 12, 2025

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Neuron Activity Dependent Redox Compartmentation Revealed with a Second Generation Red-Shifted Ratiometric Sensor.

Saranya Radhakrishnan, Jacob Norley, Stefan Wendt1

  • 1Djavad Mowafaghian Centre for Brain Health, University of British Columbia, 2215 Wesbrook Mall, Vancouver, BC V6T 1Z3, Canada.

ACS Chemical Neuroscience
|August 14, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed a new red fluorescent sensor, rogRFP2, to track cellular redox changes. This tool reveals how neuronal activity impacts mitochondria and cytosol, aiding neurological disorder research.

Keywords:
Activity-DependenceCompartmentationGenetically-Encoded Fluorescent Protein SensorMitochondriaNeuronOxidative StressRedox

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Oxidative stress is implicated in neurological disorders, but understanding subcellular redox dynamics during neuronal activity is challenging.
  • Existing redox probes lack the multicolor capabilities needed for compartment-specific analysis in single cells, often masked by population averaging.

Purpose of the Study:

  • To develop a second-generation, genetically encoded, excitation ratiometric redox-sensitive fluorescent protein sensor (rogRFP2) with improved red emission for quantitative live-cell imaging.
  • To utilize rogRFP2 to investigate activity-dependent redox changes in individual neurons and their subcellular compartments.

Main Methods:

  • Engineering of a second-generation red-shifted redox-sensitive fluorescent protein sensor (rogRFP2) using a Förster resonance energy transfer relay strategy.
  • Quantitative live-cell imaging of cultured neurons using rogRFP2 to measure compartment-specific redox dynamics.
  • Ratiometric one- and two-photon redox imaging in rat brain slices and Drosophila retinas.

Main Results:

  • Observed an anticorrelation between mitochondrial oxidation and cytosolic reduction in response to neuronal activity.
  • Demonstrated that this redox behavior is dependent on Complex I activity in the mitochondrial electron transport chain.
  • Showed that cocultured astrocytes can modulate these activity-dependent redox changes.
  • Validated rogRFP2 for ratiometric redox imaging in brain slices and fly retinas.

Conclusions:

  • The novel rogRFP2 sensor enables quantitative, compartment-specific redox imaging in live cells, brain slices, and fly retinas.
  • This tool provides new insights into the relationship between neuronal activity and subcellular redox changes.
  • rogRFP2 is a powerful tool for advancing redox biology research in vitro and in vivo across model organisms.