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Rapid optical control of nociception with an ion-channel photoswitch

Nature Methods volume 9, pages 396–402 (2012)Cite this article

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Abstract

Local anesthetics effectively suppress pain sensation, but most of these compounds act nonselectively, inhibiting activity of all neurons. Moreover, their actions abate slowly, preventing precise spatial and temporal control of nociception. We developed a photoisomerizable molecule, quaternary ammonium–azobenzene–quaternary ammonium (QAQ), that enables rapid and selective optical control of nociception. QAQ is membrane-impermeant and has no effect on most cells, but it infiltrates pain-sensing neurons through endogenous ion channels that are activated by noxious stimuli, primarily TRPV1. After QAQ accumulates intracellularly, it blocks voltage-gated ion channels in the trans form but not the cis form. QAQ enables reversible optical silencing of mouse nociceptive neuron firing without exogenous gene expression and can serve as a light-sensitive analgesic in rats in vivo. Because intracellular QAQ accumulation is a consequence of nociceptive ion-channel activity, QAQ-mediated photosensitization is a platform for understanding signaling mechanisms in acute and chronic pain.

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Figure 1: Intracellular QAQ photosensitized voltage-gated ion channels.
Figure 2: Intracellular QAQ as a photoswitchable inhibitor of neuronal activity.
Figure 3: TRPV1 channels and P2X7 receptors as a conduit for QAQ entry into cells.
Figure 4: Photosensitization of intact DRGs recorded with a three-dimensional multielectrode array (MEA).
Figure 5: Photosensitization of spinal cord slices.
Figure 6: QAQ optically regulated nociception in vivo, in live rats.

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Acknowledgements

We thank M.R. Banghart and M. Kienzler for helping synthesize QAQ, S. Scott for his help with computer programming, D. Bautista for helpful comments, A. Nicke (Max Planck Institute of Brain Research) for the P2X7 receptor clone, A. Blatz (Photoswitch Biosciences, Inc.) for HEK-293 cells stably expressing Cav2.2 and F. Ory for artistic input. This work was supported by US National Institutes of Health grants MH088484 to R.H.K. and PN2 EY018241 to R.H.K. and D.T. (the University of California Berkeley Nanomedicine Development Center) and by the Center for Integrated Protein Science, Munich to D.T.

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Author notes

  1. Alexandre Mourot and Timm Fehrentz: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California, USA

    Alexandre Mourot, Timm Fehrentz, Caleb M Smith, Christian Herold & Richard H Kramer

  2. Department of Chemistry, University of Munich, Munich, Germany

    Timm Fehrentz & Dirk Trauner

  3. Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, Unité Mixte de Recherche 5297 'Central Mechanisms of Pain Sensitization', Bordeaux, France

    Yves Le Feuvre & Frédéric Nagy

  4. Université de Bordeaux, Bordeaux, France

    Yves Le Feuvre & Frédéric Nagy

  5. Department of Chemical Engineering and Department of Bioengineering and The Helen Wills Neuroscience Institute, University of California, Berkeley, California, USA

    Deniz Dalkara

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Contributions

A.M. and R.H.K. wrote the paper. A.M., T.F., Y.L.F., C.M.S., F.N., D.T. and. R.H.K. designed experiments. AM., T.F., Y.L.F., C.M.S. and C.H. performed electrophysiological experiments and analyzed data. A.M. and D.D. performed in vivo experiments.

Corresponding authors

Correspondence to Dirk Trauner or Richard H Kramer.

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The authors declare no competing financial interests.

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Mourot, A., Fehrentz, T., Le Feuvre, Y. et al. Rapid optical control of nociception with an ion-channel photoswitch. Nat Methods 9, 396–402 (2012). https://doi.org/10.1038/nmeth.1897

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