Archaerhodopsin variants with enhanced voltage-sensitive fluorescence in mammalian and Caenorhabditis elegans neurons

Nicholas C. Flytzanis, Claire N. Bedbrook, Hui Chiu, Martin K. M. Engqvist, Cheng Xiao, Ken Y. Chan, Paul W. Sternberg, Frances H. Arnold and Viviana Gradinaru ()
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Nicholas C. Flytzanis: California Institute of Technology
Claire N. Bedbrook: California Institute of Technology
Hui Chiu: California Institute of Technology
Martin K. M. Engqvist: California Institute of Technology
Cheng Xiao: California Institute of Technology
Ken Y. Chan: California Institute of Technology
Paul W. Sternberg: California Institute of Technology
Frances H. Arnold: California Institute of Technology
Viviana Gradinaru: California Institute of Technology

Nature Communications, 2014, vol. 5, issue 1, 1-9

Abstract: Abstract Probing the neural circuit dynamics underlying behaviour would benefit greatly from improved genetically encoded voltage indicators. The proton pump Archaerhodopsin-3 (Arch), an optogenetic tool commonly used for neuronal inhibition, has been shown to emit voltage-sensitive fluorescence. Here we report two Arch variants with enhanced radiance (Archers) that in response to 655 nm light have 3–5 times increased fluorescence and 55–99 times reduced photocurrents compared with Arch WT. The most fluorescent variant, Archer1, has 25–40% fluorescence change in response to action potentials while using 9 times lower light intensity compared with other Arch-based voltage sensors. Archer1 is capable of wavelength-specific functionality as a voltage sensor under red light and as an inhibitory actuator under green light. As a proof-of-concept for the application of Arch-based sensors in vivo, we show fluorescence voltage sensing in behaving Caenorhabditis elegans. Archer1’s characteristics contribute to the goal of all-optical detection and modulation of activity in neuronal networks in vivo.

Date: 2014
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DOI: 10.1038/ncomms5894

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