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Photochemical control of endogenous ion channels and cellular excitability

Nature Methods volume 5, pages 331–338 (2008)Cite this article

Abstract

Light-activated ion channels provide a precise and noninvasive optical means for controlling action potential firing, but the genes encoding these channels must first be delivered and expressed in target cells. Here we describe a method for bestowing light sensitivity onto endogenous ion channels that does not rely on exogenous gene expression. The method uses a synthetic photoisomerizable small molecule, or photoswitchable affinity label (PAL), that specifically targets K+ channels. PALs contain a reactive electrophile, enabling covalent attachment of the photoswitch to naturally occurring nucleophiles in K+ channels. Ion flow through PAL-modified channels is turned on or off by photoisomerizing PAL with different wavelengths of light. We showed that PAL treatment confers light sensitivity onto endogenous K+ channels in isolated rat neurons and in intact neural structures from rat and leech, allowing rapid optical regulation of excitability without genetic modification.

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Figure 1: The PAL approach for imparting light sensitivity onto native ion channels.
Figure 2: Photocontrol of K+ channels expressed in HEK293 cells.
Figure 3: Photocontrol of native K+ current in cultured hippocampal neurons.
Figure 4: Photocontrol of neuronal firing.
Figure 5: Modulation of neuronal excitability with light.
Figure 6: Local illumination during PAL treatment imprints photosensitivity onto specific neurons.
Figure 7: Photocontrol of action potential firing in intact neural circuits.

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Acknowledgements

We thank E. Isacoff, J. Chambers, S.-Y. Choi and S. Jackman for helpful comments, J. Flannery and K. Greenberg for help with RGC experiments, D. Johnston (University of Texas at Austin), B. Rothberg (University of Texas Health Science Center at San Antonio), B. Rudy (New York University), W. Catterall (University of Washington) and J. Trimmer (University of California Davis) for providing plasmids. This work was supported by the Howard Hughes Medical Institute (K.B.), the US National Science Foundation (IOB-0523959 to W.B.K.), Microsoft Research Labs (to W.B.K.) and the US National Institutes of Health (GM057027 to D.T., MH43396 to W.B.K., and EY16249 to R.H.K.). D.A.W., Q.G. and W.B.K. thank A. Blankenship and M. Feller for the loan of their xenon lamp.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California Berkeley, 121 Life Sciences Addition, Berkeley, 94720, California, USA

    Doris L Fortin, Timothy W Dunn, Katharine Borges & Richard H Kramer

  2. Department of Chemistry, University of California Berkeley, Latimer Hall, Berkeley, 94720, California, USA

    Matthew R Banghart & Dirk Trauner

  3. Department of Biology, University of California San Diego, 9500 Gilman Drive, La Jolla, 92037, California, USA

    Daniel A Wagenaar, Quentin Gaudry & William B Kristan

  4. Department of Neurobiology, University of California, Los Angeles, 63-314 CHS Box 951763, Los Angeles, 90095, California, USA

    Movses H Karakossian & Thomas S Otis

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  1. Doris L Fortin
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Correspondence to Dirk Trauner or Richard H Kramer.

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Fortin, D., Banghart, M., Dunn, T. et al. Photochemical control of endogenous ion channels and cellular excitability. Nat Methods 5, 331–338 (2008). https://doi.org/10.1038/nmeth.1187

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