Universität zu Köln

Govorunova, Elena G., Sineshchekov, Oleg A., Hemmati, Raheleh, Janz, Roger ORCID: 0000-0002-9872-0596, Morelle, Olivier, Melkonian, Michael, Wong, Gane K. -S. and Spudich, John L. (2018). Extending the Time Domain of Neuronal Silencing with Cryptophyte Anion Channelrhodopsins. eNeuro, 5 (3). WASHINGTON: SOC NEUROSCIENCE. ISSN 2373-2822

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Abstract

Optogenetic inhibition of specific neuronal types in the brain enables analysis of neural circuitry and is promising for the treatment of a number of neurological disorders. Anion channelrhodopsins (ACRs) from the cryptophyte alga Guillardia theta generate larger photocurrents than other available inhibitory optogenetic tools, but more rapid channels are needed for temporally precise inhibition, such as single-spike suppression, of high-frequency firing neurons. Faster ACRs have been reported, but their potential advantages for time-resolved inhibitory optogenetics have not so far been verified in neurons. We report RapACR, nicknamed so for rapid, an ACR from Rhodomonas salina, that exhibits channel half-closing times below 10 ms and achieves equivalent inhibition at 50-fold lower light intensity in lentivirally transduced cultured mouse hippocampal neurons as the second-generation engineered Cl--conducting channelrhodopsin iC++. The upper limit of the time resolution of neuronal silencing with RapACR determined by measuring the dependence of spiking recovery after photoinhibition on the light intensity was calculated to be 100 Hz, whereas that with the faster of the two G. theta ACRs was 13 Hz. Further acceleration of RapACR channel kinetics was achieved by site-directed mutagenesis of a single residue in transmembrane helix 3 (Thr111 to Cys). We also show that mutation of another ACR (Cys to Ala at the same position) with a greatly extended lifetime of the channel open state acts as a bistable photochromic tool in mammalian neurons. These molecules extend the time domain of optogenetic neuronal silencing while retaining the high light sensitivity of Guillardia ACRs.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Govorunova, Elena G. UNSPECIFIED UNSPECIFIED UNSPECIFIED Sineshchekov, Oleg A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hemmati, Raheleh UNSPECIFIED UNSPECIFIED UNSPECIFIED Janz, Roger UNSPECIFIED orcid.org/0000-0002-9872-0596 UNSPECIFIED Morelle, Olivier UNSPECIFIED UNSPECIFIED UNSPECIFIED Melkonian, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED Wong, Gane K. -S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Spudich, John L. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-187331
DOI:	10.1523/ENEURO.0174-18.2018
Journal or Publication Title:	eNeuro
Volume:	5
Number:	3
Date:	2018
Publisher:	SOC NEUROSCIENCE
Place of Publication:	WASHINGTON
ISSN:	2373-2822
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language OPTOGENETIC INHIBITION; OPTICAL INTERROGATION; CHLORIDE CHANNEL; NEURAL CIRCUITS; BEHAVIOR; TRANSMISSION; CONSTRAINTS; MECHANISMS; STRATEGIES; TOOLS Multiple languages Neurosciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18733