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Nat Commun. 2015 Sep 8;6:8046. doi: 10.1038/ncomms9046.

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

Author information

1
Department of Biology, Institute for Molecular Plant Physiology and Biophysics, Biocenter, Julius-Maximilians-University of Würzburg, Julius-von-Sachs-Platz 2, D-97082 Würzburg, Germany.
2
Buchmann Institute for Molecular Life Sciences, Goethe University, Max von Laue Strasse 15, D-60438 Frankfurt, Germany.
3
Department for Biochemistry, Chemistry and Pharmacy, Institute of Biochemistry, Goethe University, Max von Laue Strasse 9, D-60438 Frankfurt, Germany.
4
Department of Microbiology, Biocenter, Julius-Maximilians-University of Würzburg, Am Hubland, D-97074 Würzburg, Germany.
5
Cluster of Excellence Frankfurt-Macromolecular Complexes (CEF-MC), Goethe University, Max von Laue Strasse 15, D-60438 Frankfurt, Germany.

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.

PMID:
26345128
PMCID:
PMC4569695
DOI:
10.1038/ncomms9046
[Indexed for MEDLINE]
Free PMC Article

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