Effects of Intracellular Calcium and Actin Cytoskeleton on TCR Mobility Measured by Fluorescence Recovery 英文参考文献.docVIP

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Effects of Intracellular Calcium and Actin Cytoskeleton on TCR Mobility Measured by Fluorescence Recovery 英文参考文献.doc

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Effects of Intracellular Calcium and Actin Cytoskeleton on TCR Mobility Measured by Fluorescence Recovery 英文参考文献

EffectsofIntracellularCalciumandActinCytoskeleton onTCRMobilityMeasuredbyFluorescenceRecovery OmerDushek1.,SabinaMueller2.,SebastienSoubies2¤a,DavidDepoil2¤b,IrisCaramalho2¤c ,Daniel Coombs1.,SalvatoreValitutti2. * 1Department of Mathematics and Institute of Applied Mathematics, University of British Columbia, Vancouver, British Columbia, Canada, 2INSERM, U563, Centre de PhysiopathologiedeToulousePurpan,SectionDynamiquemole′culairedesinteractionslymphocytaires,Toulouse,France Abstract Background:TheactivationofTlymphocytesbyspecificantigenisaccompaniedbytheformationofaspecializedsignaling region termed theimmunological synapse, characterized by theclustering andsegregation of surface moleculesand,in particular,byTcellreceptor(TCR)clustering. Methodology/Principal Findings: To better understand TCR motion during cellular activation, we used confocal microscopyandphoto-bleachingrecoverytechniquestoinvestigatethelateralmobilityofTCRonthesurfaceofhumanT lymphocytes under various pharmacological treatments. Using drugs that cause an increase in intracellular calcium, we observed a decrease in TCR mobility that was dependent on a functional actin cytoskeleton. In parallel experiments measurement of filamentous actin by FACS analysis showed that raising intracellular calcium also causes increased polymerizationoftheactin cytoskeleton. These invitroresults wereanalyzedusingamathematical modelthatrevealed effectivebindingparametersbetweenTCRandtheactincytoskeleton. Conclusion/Significance: We propose, based on our results, that increase in intracellular calcium levels leads to actin polymerizationandincreasesTCR/cytoskeletoninteractionsthatreducetheoverallmobilityoftheTCR.Inaphysiological setting,thismaycontributetoTCRre-positioningattheimmunologicalsynapse. Citation:DushekO,MuellerS,SoubiesS,DepoilD,CaramalhoI,etal.(2008)EffectsofIntracellularCalciumandActinCytoskeletononTCRMobilityMeasuredby FluorescenceRecovery.PLoSONE3(12):e3913.doi:10.1371/journal.pone.0003913 Edi