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Spying on Neuronal Membrane Potential with Genetically Targetable Voltage Indicators.

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    New VoltageSpy dyes enable precise optical measurement of voltage changes in genetically defined neurons. These chemically synthesized fluorescent probes can be targeted to specific cells, offering high sensitivity and fast response times for neuronal activity imaging.

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    Area of Science:

    • Neuroscience
    • Biochemistry
    • Optical Imaging

    Background:

    • Optical voltage measurement offers superior spatial and temporal resolution compared to traditional methods.
    • Existing voltage-sensitive dyes lack specific cell-targeting capabilities, limiting their application.
    • Genetically encoded voltage indicators provide cell specificity but often have slower kinetics and lower sensitivity.

    Purpose of the Study:

    • To develop a novel class of voltage-sensitive fluorescent dyes (sarcosine-VoltageFluors) that can be genetically targeted to specific cells.
    • To enable high-resolution optical imaging of voltage dynamics in genetically defined neurons, including axons and dendrites.
    • To combine the speed and sensitivity of synthetic dyes with the cellular specificity of genetic probes.

    Main Methods:

    • Synthesis of fluorescein-based voltage-sensitive fluorescent dyes with functional carboxylic acid groups.
    • Conjugation of dyes to a polyethylene glycol (PEG) linker and a SpyTag peptide for specific binding to SpyCatcher.
    • Expression of SpyCatcher on the cell surface of neurons for targeted dye delivery.
    • Optical imaging of action potentials in cultured neurons using the VoltageSpy system.

    Main Results:

    • The new VoltageSpy dyes effectively labeled cells expressing cell-surface SpyCatcher.
    • VoltageSpy dyes demonstrated good voltage sensitivity and fast-response kinetics, exceeding genetically encoded voltage indicators.
    • Robust, single-trial optical detection of action potentials was achieved in neuronal soma, axons, and dendrites.
    • Voltage imaging was successful at low nanomolar dye concentrations.

    Conclusions:

    • VoltageSpy represents a significant advancement in voltage imaging, enabling genetically targeted optical measurement of neuronal electrical activity.
    • This technology merges the advantages of synthetic voltage sensors and genetic targeting, paving the way for detailed studies of neuronal function.
    • VoltageSpy offers a powerful tool for neuroscience research, allowing precise visualization of electrical signals in specific neuronal populations.