Abstract
Genetically targeted light-activated ion channels and pumps make it possible to determine the role of specific neurons in neuronal circuits, information processing and behavior. We developed a K+-selective ionotropic glutamate receptor that reversibly inhibits neuronal activity in response to light in dissociated neurons and brain slice and also reversibly suppresses behavior in zebrafish. The receptor is a chimera of the pore region of a K+-selective bacterial glutamate receptor and the ligand-binding domain of a light-gated mammalian kainate receptor. This hyperpolarizing light-gated channel, HyLighter, is turned on by a brief light pulse at one wavelength and turned off by a pulse at a second wavelength. The control is obtained at moderate intensity. After optical activation, the photocurrent and optical silencing of activity persists in the dark for extended periods. The low light requirement and bi-stability of HyLighter represent advantages for the dissection of neural circuitry.
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Acknowledgements
We thank K. Partin, K. Kainanen, E. Gouaux and M. Hollmann for constructs, G. Sandoz, P. Koprowski and F. Tombola for discussions, D. Fortin, T. Tracey, K. Greenberg, A. Pham, M. Soden, H. Lu, E. Warp, K. McDaniel, R. Arant and Z. Fu for technical assistance, and M. Volgraf and V. Franckevicius for MAG compounds. This work was supported by the US National Institutes of Health Nanomedicine Development Center for the Optical Control of Biological Function (5PN2EY018241), the Human Frontier Science Program (RPG23-2005), the National Science Foundation (FIBR 43C-1081892), a fellowship of the European Molecular Biology Organization (to H.J.) and a Marie-Curie fellowship (to C.W.; the fellowship was obtained with the laboratory CNRS-UMR5020 'Neurosciences Sensorielles, Comportement Cognition').
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H.J. designed chimeric proteins, conducted experiments in HEK293 cells and neuronal cultures, contributed to experiments in brain slices and zebrafish, analyzed data and wrote the manuscript. S.S. conducted experiments in brain slices, analyzed data and wrote the manuscript. C.W. conducted experiments in zebrafish, analyzed data and wrote the manuscript. D.T. developed photoswitching methodology and provided photoswitches. E.Y.I. designed chimeric proteins, developed photoswitching methodology, analyzed data and wrote the manuscript.
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Janovjak, H., Szobota, S., Wyart, C. et al. A light-gated, potassium-selective glutamate receptor for the optical inhibition of neuronal firing. Nat Neurosci 13, 1027–1032 (2010). https://doi.org/10.1038/nn.2589
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DOI: https://doi.org/10.1038/nn.2589
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