Simultaneous Detection of Ca2+ and Diacylglycerol Signaling in Living Cells 英文参考文献.docVIP

SimultaneousDetectionofCa2+andDiacylglycerol SignalinginLivingCells PaulTewson1,MaraWestenberg1,YongxinZhao3,RobertE.Campbell3,AnneMarieQuinn1, ThomasE.Hughes1,2 * 1Montana Molecular, Bozeman, Montana, United States of America, 2Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana, UnitedStatesofAmerica,3DepartmentofChemistry,UniversityofAlberta,Edmonton,Alberta,Canada Abstract Phospholipase C produces twosecond messengers -diacylglycerol (DAG), which remains in themembrane, andinositol triphosphate(IP3),whichtriggersthereleaseofcalciumions(Ca )fromintracellularstores.Geneticallyencodedsensors basedonasinglecircularlypermutedfluorescentprotein(FP)arerobusttoolsforstudyingintracellularCa dynamics.We 2+ 2+ have developed a robust sensor for DAG based on a circularly permuted green FP that can be co-imaged with the red fluorescentCa sensorR-GECOforsimultaneousmeasurementofbothsecondmessengers. 2+ Citation:TewsonP,WestenbergM,ZhaoY,CampbellRE,QuinnAM,etal.(2012)SimultaneousDetectionofCa2+andDiacylglycerolSignalinginLivingCells.PLoS ONE7(8):e42791.doi:10.1371/journal.pone.0042791 Editor:AlexanderG.Obukhov,IndianaUniversitySchoolofMedicine,UnitedStatesofAmerica ReceivedJune4,2012;AcceptedJuly10,2012;PublishedAugust17,2012 Copyright: ? 2012 Tewson et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding: This work was supported by the NIMH - National Institue of Mental Health (1R43MH096670- 01A1), and The Montana Board of Research and Commercialization #11-37 to A.M.Q., and by Alberta Innovates (Y.Z.), Canadian Institutes of Health Research (R.E.C), and Natural Sciences and Engineering ResearchCouncilofCanada(R.E.C.).R.E.C.holdsaTierIICanadaResearchChairinBioanalyticalChemistry. CompetingInterests:PT,MW,AMQ,andTHareemployedbyMontanaMolecularandhav