原创力文档reproducible and sustained regulation of gαs signalling using a metazoan opsin as an optogenetic tool可再生的和持续的监管gαs信号用一个后生动物视蛋白作为optogenetic工具.pdfVIP
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reproducible and sustained regulation of gαs signalling using a metazoan opsin as an optogenetic tool可再生的和持续的监管gαs信号用一个后生动物视蛋白作为optogenetic工具
Reproducible and Sustained Regulation of Gas Signalling
Using a Metazoan Opsin as an Optogenetic Tool
Helena J. Bailes, Ling-Yu Zhuang, Robert J. Lucas*
Faculty of Life Sciences, The University of Manchester, Manchester, United Kingdom
Abstract
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 2
mW/cm . We conclude that JellyOp is a promising
new tool for mimicking the activity of Gs-coupled G protein coupled receptors with fine spatiotemporal resolution.
Citation: Bailes HJ, Zhuang L-Y, Lucas RJ (2012) Reproducible and Sustained Regulation of Gas Signalling Using a Metazoan Opsin as an Optogenetic Tool. PLoS
ONE 7(1): e30774. doi:10.1371/journal.pone.0030774
Editor: Paul A. Bartell
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