Synthetic retinal analogues modify the spectral and kinetic characteristics of microbial rhodopsin optogenetic tools

Optogenetic tools have become indispensable in neuroscience to stimulate or inhibit excitable cells by light. Channelrhodopsin-2 (ChR2) variants have been established by mutating the opsin backbone or by mining related algal genomes. As an alternative strategy, we surveyed synthetic retinal analogue...

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Vydáno v:Nature communications Ročník 5; číslo 1; s. 5810
Hlavní autoři: AzimiHashemi, N., Erbguth, K., Vogt, A., Riemensperger, T., Rauch, E., Woodmansee, D., Nagpal, J., Brauner, M., Sheves, M., Fiala, A., Kattner, L., Trauner, D., Hegemann, P., Gottschalk, A., Liewald, J. F.
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 15.12.2014
Nature Publishing Group
Témata:
13/106
14/63
631/1647/2253
631/378/2586
631/92/436
64/11
64/24
9/74
Action Potentials - physiology
Algae
Animals
Animals, Genetically Modified
Behavior, Animal
Caenorhabditis elegans - chemistry
Caenorhabditis elegans - drug effects
Caenorhabditis elegans - metabolism
Drosophila melanogaster - chemistry
Drosophila melanogaster - drug effects
Drosophila melanogaster - metabolism
Drosophila Proteins - chemistry
Genomes
HEK293 Cells
Helminth Proteins - chemistry
Humanities and Social Sciences
Humans
Insects
Larva - chemistry
Larva - drug effects
Larva - metabolism
Light
Light intensity
multidisciplinary
Neurosciences
Optogenetics - instrumentation
Patch-Clamp Techniques
Proteins
Recombinant Proteins - chemistry
Retinaldehyde - chemistry
Retinaldehyde - pharmacology
Rhodopsin - chemistry
Rhodopsins, Microbial - chemistry
Science
Science (multidisciplinary)
Wavelengths
Germany
ISSN:2041-1723, 2041-1723
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Shrnutí:Optogenetic tools have become indispensable in neuroscience to stimulate or inhibit excitable cells by light. Channelrhodopsin-2 (ChR2) variants have been established by mutating the opsin backbone or by mining related algal genomes. As an alternative strategy, we surveyed synthetic retinal analogues combined with microbial rhodopsins for functional and spectral properties, capitalizing on assays in C. elegans , HEK cells and larval Drosophila . Compared with all- trans retinal (ATR), Dimethylamino-retinal (DMAR) shifts the action spectra maxima of ChR2 variants H134R and H134R/T159C from 480 to 520 nm. Moreover, DMAR decelerates the photocycle of ChR2(H134R) and (H134R/T159C), thereby reducing the light intensity required for persistent channel activation. In hyperpolarizing archaerhodopsin-3 and Mac, naphthyl-retinal and thiophene-retinal support activity alike ATR, yet at altered peak wavelengths. Our experiments enable applications of retinal analogues in colour tuning and altering photocycle characteristics of optogenetic tools, thereby increasing the operational light sensitivity of existing cell lines or transgenic animals. Efforts to improve the performance of optogenetic tools for neuroscience research have mostly been focused on mutating the opsin backbones or mining-related algal genomes. Here the authors show that analogues of the chromophore, retinal, can be used for colour tuning of rhodopsins and altering their photocycle kinetics in several model organisms.
Bibliografie:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:2041-1723
2041-1723
DOI:10.1038/ncomms6810