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Han X., Boyden E. S. (2007) Multiple-Color Optical Activation, Silencing, and Desynchronization of Neural Activity, with Single-Spike Temporal Resolution, PLoS ONE, 2(3):e299. doi:10.1371/journal.pone.0000299.
This work was first publicly presented on 2/24/07, at a Spotlight Presentation at the CoSyNe meeting in Salt Lake City, UT, and published on 3/21/07.
Multiple-color optical activation, silencing, and desynchronization of neural activity, with single-spike temporal resolution
AbstractThe quest to determine how precise neural activity patterns mediate computation, behavior, and pathology would be greatly aided by a set of tools for reliably activating and inactivating genetically-targeted neurons, in a temporally-precise and rapidly-reversible fashion. Having earlier adapted a light-activated cation channel, channelrhodopsin-2 (ChR2), for allowing neurons to be stimulated by blue light, we searched for a complementary tool that would enable optical neuronal inhibition, driven by light of a second color.
Here we report that targeting the codon-optimized form of the light-driven chloride pump halorhodopsin from the archaebacterium Natronomas pharaonis (hereafter abbreviated Halo) to genetically-specified neurons enables them to be silenced reliably, and reversibly, by millisecond-timescale pulses of yellow light. We show that trains of yellow and blue light pulses can drive high-fidelity sequences of hyperpolarizations and depolarizations in neurons simultaneously expressing yellow light-driven Halo and blue light-driven ChR2, allowing for the first time manipulations of neural synchrony without perturbation of other parameters such as spiking rates.
The Halo/ChR2 system thus constitutes a powerful toolbox for multichannel photoinhibition and photostimulation of virally- or transgenically-targeted neural circuits without need for exogenous chemicals, enabling systematic analysis and engineering of the brain, and quantitative bioengineering of excitable cells.
Lentivirus plasmid maps and sequences:
Halorhodopsin/Halo/NpHR, with mammalian optimized codon usage, fused with GFP (FCK-Halo-GFP, containing Halo-GFP) [ plasmid map ] [ sequences for gene and plasmid ] [ plasmid data in VectorNTI format ]
Email : with general questions, or to have your study added to the list below.
Mailing list: for users of channelrhodopsin-2 and halorhodopsin (for discussing reagents, vectors, hardware, etc.). A channelrhodopsin/channelopsin/ChR2/chop-2/halorhodopsin/NPhR/Halo/photostimulation community.
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