Endoplasmic Reticulum Redox State Is Not Perturbed by Pharmacological or Pathological Endoplasmic Reticulum Stress in Live Pancreatic β-Cells 英文参考文献.docVIP

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Endoplasmic Reticulum Redox State Is Not Perturbed by Pharmacological or Pathological Endoplasmic Reticulum Stress in Live Pancreatic β-Cells 英文参考文献.doc

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Endoplasmic Reticulum Redox State Is Not Perturbed by Pharmacological or Pathological Endoplasmic Reticulum Stress in Live Pancreatic β-Cells 英文参考文献

EndoplasmicReticulumRedoxStateIsNotPerturbedby PharmacologicalorPathologicalEndoplasmicReticulum StressinLivePancreaticb-Cells IrmgardSchuiki1,LilingZhang1,AllenVolchuk1,2,3* 1Division of Cellular and Molecular Biology, Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada, 2Department of Biochemistry, UniversityofToronto,Toronto,Ontario,Canada,3DepartmentofPhysiology,UniversityofToronto,Toronto,Ontario,Canada Abstract Accumulationofunfolded,misfoldedandaggregatedproteinsintheendoplasmicreticulum(ER)causesERstress.ERstress canresultfromphysiologicalsituationssuchasacuteincreasesinsecretoryproteinbiosynthesisorpathologicalconditions that perturb ER homeostasis such as alterations in the ER redox state. Here we monitored ER redox together with transcriptionaloutputoftheUnfoldedProteinResponse(UPR)inINS-1insulinomacellsstablyexpressingeroGFP(ER-redox- sensor)andmCherryproteindrivenbyaGRP78promoter(UPR-sensor).Livecellimaging,flowcytometryandbiochemical characterizationwereusedtoexaminetheseparametersinresponsetovariousconditionsknowntoinduceERstress.As expected,treatmentofthecellswiththereducingagentdithiothreitolcausedadecreaseintheoxidationstateoftheER accompanied by an increase in XBP-1 splicing. Unexpectedly however, other treatments including tunicamycin, thapsigargin, DL-homocysteine, elevated free fatty acids or high glucose had essentially no influence on the ER redox state,despiteinducingERstress.ComparableresultswereobtainedwithdispersedratisletcellsexpressingeroGFP.Thus, unlikeinyeastcells,ERstressinpancreaticb-cellsisnotassociatedwithamorereducingERenvironment. Citation:SchuikiI,ZhangL,VolchukA(2012)EndoplasmicReticulumRedoxStateIsNotPerturbedbyPharmacologicalorPathologicalEndoplasmicReticulum StressinLivePancreaticb-Cells.PLoSONE7(11):e48626.doi:10.1371/journal.pone.0048626 Editor:JeffreyL.Brodsky,UniversityofPittsburgh,UnitedStatesofAmerica ReceivedJuly13,2012;AcceptedOctober3,2012;PublishedNovember8,2012 Copyright: ? 20