Nature Methods
- 5, 331 - 338 (2008)
Published online: 2 March 2008; | doi:10.1038/nmeth.1187
Photochemical control of endogenous ion channels and cellular excitabilityDoris L Fortin1, Matthew R Banghart2, Timothy W Dunn1, Katharine Borges1, Daniel A Wagenaar3, Quentin Gaudry3, Movses H Karakossian4, Thomas S Otis4, William B Kristan3, Dirk Trauner2 & Richard H Kramer11
Department of Molecular and Cell Biology, University of California Berkeley, 121 Life Sciences Addition, Berkeley, California 94720, USA. 2
Department of Chemistry, University of California Berkeley, Latimer Hall, Berkeley, California 94720, USA. 3
Department of Biology, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92037, USA. 4
Department of Neurobiology, University of California, Los Angeles, 63-314 CHS Box 951763, Los Angeles, California 90095, USA.
Correspondence should be addressed to Dirk Trauner trauner@berkeley.edu or Richard H Kramer rhkramer@berkeley.edu 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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