Optogenetic manipulation of cGMP in cells and animals by the tightly light-regulated guanylyl-cyclase opsin CyclOp

Nature Communications
Shiqiang GaoAlexander Gottschalk

Abstract

Cyclic GMP (cGMP) signalling regulates multiple biological functions through activation of protein kinase G and cyclic nucleotide-gated (CNG) channels. In sensory neurons, cGMP permits signal modulation, amplification and encoding, before depolarization. Here we implement a guanylyl cyclase rhodopsin from Blastocladiella emersonii as a new optogenetic tool (BeCyclOp), enabling rapid light-triggered cGMP increase in heterologous cells (Xenopus oocytes, HEK293T cells) and in Caenorhabditis elegans. Among five different fungal CyclOps, exhibiting unusual eight transmembrane topologies and cytosolic N-termini, BeCyclOp is the superior optogenetic tool (light/dark activity ratio: 5,000; no cAMP production; turnover (20 °C) ∼17 cGMP s(-1)). Via co-expressed CNG channels (OLF in oocytes, TAX-2/4 in C. elegans muscle), BeCyclOp photoactivation induces a rapid conductance increase and depolarization at very low light intensities. In O2/CO2 sensory neurons of C. elegans, BeCyclOp activation evokes behavioural responses consistent with their normal sensory function. BeCyclOp therefore enables precise and rapid optogenetic manipulation of cGMP levels in cells and animals.

References

Nov 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·W AltenhofenU B Kaupp
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·E N SpudichJ L Spudich
Sep 29, 1971·Nature: New Biology·D Oesterhelt, W Stoeckenius
Jun 29, 2002·Science·Georg NagelPeter Hegemann
Dec 10, 2002·Annual Review of Physiology·Sławomir FilipekKrzysztof Palczewski
Oct 25, 2003·Cellular Signalling·Jürgen U Linder, Joachim E Schultz
Nov 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Georg NagelErnst Bamberg
Apr 28, 2004·Journal of Molecular Biology·Lukas KällErik L L Sonnhammer
May 15, 2004·News in Physiological Sciences : an International Journal of Physiology Produced Jointly by the International Union of Physiological Sciences and the American Physiological Society·Suneel KateriyaPeter Hegemann
Jun 3, 2004·Genome Research·Gavin E CrooksSteven E Brenner
Aug 24, 2005·Nature Neuroscience·Edward S BoydenKarl Deisseroth
Jun 21, 2006·Cellular and Molecular Life Sciences : CMLS·J U Linder
Nov 28, 2006·Nature Methods·Saskia Schröder-LangGeorg Nagel
Apr 6, 2007·Nature·Feng ZhangKarl Deisseroth
Sep 12, 2007·Bioinformatics·M A LarkinD G Higgins
May 6, 2008·Vision Research·Theodore G Wensel
Jun 6, 2008·Proceedings of the National Academy of Sciences of the United States of America·Andrew Jonathan BretscherMario de Bono
Jun 6, 2008·Proceedings of the National Academy of Sciences of the United States of America·Elissa A Hallem, Paul W Sternberg
Jul 29, 2008·Nature Neuroscience·Daniel RamotMiriam B Goodman
Sep 17, 2008·Nature Methods·Jana F LiewaldAlexander Gottschalk
Sep 2, 2009·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Yoshinori Shichida, Take Matsuyama
Aug 10, 2010·Current Opinion in Neurobiology·Christian BamannErnst Bamberg
Oct 30, 2010·The Journal of Biological Chemistry·Min-Hyung RyuMark Gomelsky
Nov 12, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Thomas KnöpfelEdward S Boyden
Feb 1, 2011·Nature Methods·Stefanie RedemannHenrik Bringmann
Jun 7, 2011·Nature Methods·Nicholas A SwierczekRex A Kerr
Jun 23, 2011·Annual Review of Neuroscience·Lief FennoKarl Deisseroth
Dec 27, 2011·Cell·Feng ZhangKarl Deisseroth
Feb 22, 2012·Progress in Brain Research·Guillaume P DuguéThomas Knöpfel
Apr 7, 2012·Biological Psychiatry·Yuan Mei, Feng Zhang
Oct 16, 2012·Science·Roger C Hardie, Kristian Franze
Jun 27, 2013·Nature Neuroscience·Richard H KramerHillel Adesnik
May 20, 2014·Current Biology : CB·Gabriela M AvelarSuely L Gomes
Jun 4, 2014·Proceedings of the National Academy of Sciences of the United States of America·Carlos GasserAndreas Möglich

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Citations

Sep 18, 2016·Current Opinion in Chemical Biology·Brian RossJin Zhang
Mar 17, 2017·Annual Review of Biochemistry·Elena G GovorunovaJohn L Spudich
Nov 25, 2017·The Journal of Experimental Biology·Andrew J M Swafford, Todd H Oakley
Dec 1, 2017·Nature Reviews. Gastroenterology & Hepatology·Werend BoesmansPieter Vanden Berghe
Jul 13, 2018·PLoS Pathogens·Ruben Dario Arroyo-OlarteNishith Gupta
Mar 23, 2017·Journal of Bacteriology·Min-Hyung RyuMark Gomelsky
Nov 10, 2017·The Journal of Biological Chemistry·Ramasamy P KumarDaniel D Oprian
May 9, 2019·Photochemistry and Photobiology·Yumeka YamauchiHideki Kandori
Aug 10, 2019·Protein Science : a Publication of the Protein Society·Andreas Möglich
Dec 7, 2019·The FEBS Journal·Agata ButrynAllen M Orville
Nov 30, 2019·The Plant Journal : for Cell and Molecular Biology·Eva L von der Heyde, Armin Hallmann
Oct 20, 2018·Frontiers in Neuroscience·Sebastian BeckShiqiang Gao
Feb 15, 2019·Biological Chemistry·Birthe StüvenAndreas Möglich
Mar 18, 2017·The Journal of Biological Chemistry·Kazuho YoshidaHideki Kandori
Oct 18, 2020·International Journal of Molecular Sciences·Yuehui TianShiqiang Gao
Jul 14, 2019·Journal of Molecular Biology·Robert StabelAndreas Möglich
Mar 14, 2019·Biological Chemistry·Birthe StüvenAndreas Möglich
May 20, 2020·ACS Omega·Masahiro SugiuraHideki Kandori
Jul 10, 2018·Chemical Reviews·Aba LosiAndreas Möglich
Dec 10, 2019·Biochemistry·Keiichi KojimaYuki Sudo
Aug 4, 2021·Annual Review of Microbiology·Andrey RozenbergOded Béjà
Nov 3, 2017·Neuron·Benjamin R RostPeter Hegemann

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Datasets Mentioned

BETA
KF309499.1

Methods Mentioned

BETA
transfection
Fluorescence
dissection
transgenic
PCR
FCS
fluorescence imaging

Software Mentioned

LabView
BoxShade
TMHMM
BeCyclOp
manager
LabView script
Micro
LSM
CyclOp
CaCyclOp

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