Extending the Time Domain of Neuronal Silencing with Cryptophyte Anion Channelrhodopsins

ENeuro
Elena G GovorunovaJohn L Spudich

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...Continue Reading

Citations

Sep 3, 2020·Proceedings of the National Academy of Sciences of the United States of America·Elena G GovorunovaJohn L Spudich
Nov 29, 2020·Biochimica Et Biophysica Acta. Bioenergetics·Masaki TsujimuraHiroshi Ishikita
Jul 23, 2020·The Journal of Physical Chemistry Letters·Keiichi KojimaYuki Sudo
Aug 18, 2021·Nature Neuroscience·Hillel Adesnik, Lamiae Abdeladim

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Methods Mentioned

BETA
transfection

Software Mentioned

Clampex
ZipACR
Clampfit
GenScript
pClamp
Addgene
ClustalW
BrainBits
ImageJ
Origin

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