Reproducible and sustained regulation of Gαs signalling using a metazoan opsin as an optogenetic tool

Originally developed to regulate neuronal excitability, optogenetics is increasingly also used to control other cellular processes with unprecedented spatiotemporal resolution. Optogenetic modulation of all major G-protein signalling pathways (Gq, Gi and Gs) has been achieved using variants of mamma...

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Bibliographic Details
Published in:PloS one 2012, Vol.7 (1), p.e30774-e30774
Main Authors: Bailes, Helena J, Zhuang, Ling-Yu, Lucas, Robert J
Format: Article
Language:English
Subjects:
Adenosine
Amino Acid Sequence
Animals
Biology
Biosensing Techniques
Bleaches
Bleaching
Cells, Cultured
Cloning
Cubozoa
Cyclic AMP
Cyclic AMP - metabolism
Cyclic AMP - pharmacology
Drosophila
Excitability
G protein-coupled receptors
Gene expression
Genetics
HEK293 Cells
Humans
Information processing
Insects
Insulin
Kinases
Life sciences
Light
Light effects
Mammalian cells
Mammals
Mimicry
Molecular Sequence Data
Neurosciences - methods
Opsins - metabolism
Opsins - physiology
Optics
Phosphorylation
Photosensitivity
Pigments
Proteins
Receptors
Receptors, G-Protein-Coupled - metabolism
Receptors, G-Protein-Coupled - physiology
Reproducibility of Results
Sequence Homology, Amino Acid
Signal Transduction
Signaling
Validation Studies as Topic
White light
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Summary:Originally developed to regulate neuronal excitability, optogenetics is increasingly also used to control other cellular processes with unprecedented spatiotemporal resolution. Optogenetic modulation of all major G-protein signalling pathways (Gq, Gi and Gs) has been achieved using variants of mammalian rod opsin. We show here that the light response driven by such rod opsin-based tools dissipates under repeated exposure, consistent with the known bleaching characteristics of this photopigment. We continue to show that replacing rod opsin with a bleach resistant opsin from Carybdea rastonii, the box jellyfish, (JellyOp) overcomes this limitation. Visible light induced high amplitude, reversible, and reproducible increases in cAMP in mammalian cells expressing JellyOp. While single flashes produced a brief cAMP spike, repeated stimulation could sustain elevated levels for 10s of minutes. JellyOp was more photosensitive than currently available optogenetic tools, responding to white light at irradiances ≥1 µW/cm(2). We conclude that JellyOp is a promising new tool for mimicking the activity of Gs-coupled G protein coupled receptors with fine spatiotemporal resolution.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0030774