Journal
NATURE METHODS
Volume 5, Issue 4, Pages 331-338Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.1187
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Funding
- Howard Hughes Medical Institute Funding Source: Medline
- NEI NIH HHS [R01 EY015514, P30 EY003176, R01 EY018957-02, P30 EY003176-26A2, R01 EY018957, R21 EY016249, EY16249, R01 EY018957-01A1, R21 EY016249-02] Funding Source: Medline
- NIGMS NIH HHS [GM057027] Funding Source: Medline
- NIMH NIH HHS [R01 MH043396, MH43396] Funding Source: Medline
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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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