Optogenetics—the use of optically activated proteins to control cell function—allows for control of neurons with an unprecedented degree of spatial, temporal, and neurochemical precision. Three protocols are presented in this unit describing the use of channelrhodopsin-2 (ChR2), a light-activated cation channel. These protocols emphasize practical issues of working with ChR2, including guidelines for selecting a gene delivery method, light source, and method of tissue implantation, as well as steps for fabricating fiber optic patch cables and chronic implantable optical fibers. The first protocol describes the use of ChR2 in electrophysiological recordings from brain slices. The second and third involve the use of ChR2 in vivo, with light delivered through chronic fiber implants or guide cannula. Curr. Protoc. Neurosci. 58:2.16.1-2.16.19. © 2012 by John Wiley & Sons, Inc.
Neural substrates of awakening probed with optogenetic control of hypocretin neurons.
Temporally precise in vivo control of intracellular signalling.
In vivo light-induced activation of neural circuitry in transgenic mice expressing channelrhodopsin-2.
Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje cells.
J. Biol. Chem.
A user's guide to channelrhodopsin variants: Features, limitations and future developments.
Efficient gene transfer into the embryonic mouse brain using in vivo electroporation.
Neocortical excitation/inhibition balance in information processing and social dysfunction.
Cell type-specific channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function.
Web site for the University of North Carolina at Chapel Hill Gene Therapy Center; offers packaged virus for sale and related services.
Web site for the laboratory of Dr. Karl Deisseroth; contains information on ChR2 (and variants) DNA sequences, hardware, and protocols.
Web site for the laboratory of Dr. Ed Boyden; contains information on inhibitory optogenetic proteins.
Web site for the laboratory of Dr. Garret Stuber; contains protocols for optical fiber construction and optogenetics hardware.
Calculator for the spread of light in brain tissue, courtesy of the laboratory of Dr. Karl Deisseroth.
“Guide to Connectorization and Polishing Optical Fibers,” courtesy of Thorlabs.