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A hybrid approach to measuring electrical activity in genetically specified neurons

Nature Neuroscience volume 8, pages 16191626 (2005) | Download Citation

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Abstract

The development of genetically encoded fluorescent voltage probes is essential to image electrical activity from neuronal populations. Previous green fluorescent protein (GFP)-based probes have had limited success in recording electrical activity of neurons because of their low sensitivity and poor temporal resolution. Here we describe a hybrid approach that combines a genetically encoded fluorescent probe (membrane-anchored enhanced GFP) with dipicrylamine, a synthetic voltage-sensing molecule that partitions into the plasma membrane. The movement of the synthetic voltage sensor is translated via fluorescence resonance energy transfer (FRET) into a large fluorescence signal (up to 34% change per 100 mV) with a fast response and recovery time (0.5 ms). Using this two-component approach, we were able to optically record action potentials from neuronal cell lines and trains of action potentials from primary cultured neurons. This hybrid approach may form the basis for a new generation of protein-based voltage probes.

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Acknowledgements

We thank W. Hubbell (UCLA) for the gift of DPA and A. Charles (UCLA) for the GT1 cells. Thanks to F. Chow for her assistance with primary neuronal cultures and transfections. We also thank T. Otis and M. Pratap for preparing viruses for slice transfections and for access to their setup. This work was supported by grants from the US National Institutes of Health (GM30376 to F.B., NS30549 to I.M., NS41317 to F.E.S.), the American Heart Association (0225006Y and 0535214N to B.C.) and Deutschen Forschungsgemeinschaft (BL538-1/1 to R.B.).

Author information

Author notes

    • Baron Chanda
    •  & Rikard Blunck

    These authors contributed equally to this work.

Affiliations

  1. Departments of Physiology and Anesthesiology, David Geffen School of Medicine at the University of California Los Angeles (UCLA), Los Angeles, California 90095, USA.

    • Baron Chanda
    • , Rikard Blunck
    •  & Francisco Bezanilla
  2. Department of Neurology, David Geffen School of Medicine at the University of California Los Angeles (UCLA), Los Angeles, California 90095, USA.

    • Leonardo C Faria
    •  & Istvan Mody
  3. Department of Neurobiology, David Geffen School of Medicine at the University of California Los Angeles (UCLA), Los Angeles, California 90095, USA.

    • Felix E Schweizer

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Francisco Bezanilla.

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DOI

https://doi.org/10.1038/nn1558

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