Document Detail


Reproducible and sustained regulation of Gαs signalling using a metazoan opsin as an optogenetic tool.
MedLine Citation:
PMID:  22292038     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Helena J Bailes; Ling-Yu Zhuang; Robert J Lucas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-01-24
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-01-31     Completed Date:  2012-06-04     Revised Date:  2014-02-20    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e30774     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Biosensing Techniques
Cells, Cultured
Cubozoa
Cyclic AMP / metabolism,  pharmacology
HEK293 Cells
Humans
Molecular Sequence Data
Neurosciences / methods*
Opsins / metabolism,  physiology*
Receptors, G-Protein-Coupled / metabolism*,  physiology
Reproducibility of Results
Sequence Homology, Amino Acid
Signal Transduction
Validation Studies as Topic
Grant Support
ID/Acronym/Agency:
G0801731//Medical Research Council; //Biotechnology and Biological Sciences Research Council; //Medical Research Council
Chemical
Reg. No./Substance:
0/Opsins; 0/Receptors, G-Protein-Coupled; E0399OZS9N/Cyclic AMP
Comments/Corrections

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