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Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
09:57

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

Improved probes for hybrid voltage sensor imaging.

Dongsheng Wang1, Zhen Zhang, Baron Chanda

  • 1Department of Physiology, University of Wisconsin, Madison, WI, USA.

Biophysical Journal
|October 7, 2010
PubMed
Summary
This summary is machine-generated.

Researchers developed an improved hybrid voltage sensor (hVoS 2.0) for imaging electrical activity. This new probe significantly enhances fluorescence signals, enabling clearer visualization of voltage changes in targeted cells and neurons.

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Area of Science:

  • Neuroscience
  • Biophysics
  • Molecular Biology

Background:

  • Hybrid voltage sensors (hVoS) utilize fluorescence resonance energy transfer (FRET) to report membrane potential.
  • Existing hVoS probes have limitations in signal strength and signal-to-noise ratio for imaging cellular electrical activity.
  • Optimizing probe components is crucial for enhancing the sensitivity and reliability of voltage sensing.

Purpose of the Study:

  • To systematically engineer and optimize hybrid voltage sensors (hVoS) for improved signal transduction of membrane potential changes.
  • To enhance the normalized fluorescence change (ΔF/F) and signal-to-noise ratio (SNR) of hVoS probes.
  • To develop a superior voltage sensor for imaging electrical activity in genetically targeted cells and neurons.

Main Methods:

  • Systematic variation of fluorescent protein optical properties, membrane targeting motifs, and protein linkages.
  • Construction and testing of novel hVoS constructs, including GAP43-CerFP-t-h-ras (hVoS 2.0).
  • Quantitative assessment of probe performance using ΔF/F and SNR measurements in PC12 cells and cultured hippocampal neurons.

Main Results:

  • The optimized hVoS 2.0 construct (GAP43-CerFP-t-h-ras) demonstrated a threefold increase in ΔF/F compared to the original probe.
  • A fivefold greater signal-to-noise ratio was achieved with hVoS 2.0, reaching 70% of the theoretical optimum.
  • hVoS 2.0 successfully detected single action potentials in cultured hippocampal neurons with clear fluorescence changes in a single trial.

Conclusions:

  • The developed hVoS 2.0 represents a significant advancement in voltage sensing technology.
  • This enhanced probe provides improved sensitivity and signal quality for imaging electrical activity in genetically defined neuronal populations.
  • hVoS 2.0 is a valuable tool for neuroscience research, facilitating the study of neuronal function and electrical signaling.