Rhodopsin-based voltage imaging tools for use in muscles and neurons of Caenorhabditis elegans

Proceedings of the National Academy of Sciences of the United States of America
Negin Azimi HashemiAlexander Gottschalk

Abstract

Genetically encoded voltage indicators (GEVIs) based on microbial rhodopsins utilize the voltage-sensitive fluorescence of all-trans retinal (ATR), while in electrochromic FRET (eFRET) sensors, donor fluorescence drops when the rhodopsin acts as depolarization-sensitive acceptor. In recent years, such tools have become widely used in mammalian cells but are less commonly used in invertebrate systems, mostly due to low fluorescence yields. We systematically assessed Arch(D95N), Archon, QuasAr, and the eFRET sensors MacQ-mCitrine and QuasAr-mOrange, in the nematode Caenorhabditis elegans ATR-bearing rhodopsins reported on voltage changes in body wall muscles (BWMs), in the pharynx, the feeding organ [where Arch(D95N) showed approximately 128% ΔF/F increase per 100 mV], and in neurons, integrating circuit activity. ATR fluorescence is very dim, yet, using the retinal analog dimethylaminoretinal, it was boosted 250-fold. eFRET sensors provided sensitivities of 45 to 78% ΔF/F per 100 mV, induced by BWM action potentials, and in pharyngeal muscle, measured in simultaneous optical and sharp electrode recordings, MacQ-mCitrine showed approximately 20% ΔF/F per 100 mV. All sensors reported differences in muscle depolarization induced by...Continue Reading

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Citations

May 28, 2020·The FEBS Journal·Brandon J Berry, Andrew P Wojtovich
Aug 9, 2020·Frontiers in Neuroscience·Sebastian OltmannsDavide Raccuglia
Dec 29, 2019·International Journal of Molecular Sciences·Alfons PenzkoferPeter Hegemann
Sep 3, 2020·Proceedings of the National Academy of Sciences of the United States of America·Elena G GovorunovaJohn L Spudich

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Methods Mentioned

BETA
FRET
Fluorescence
dissection
Transgenic

Software Mentioned

R
MacQ
Archon
mCitrine
KNIME
mOrange
DMAR
QuasAr
GEVIs
Excel

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