Analysis of Actin FLAP Dynamics in the Leading Lamella 英文参考文献.docVIP

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Analysis of Actin FLAP Dynamics in the Leading Lamella 英文参考文献.doc

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Analysis of Actin FLAP Dynamics in the Leading Lamella 英文参考文献

AnalysisofActinFLAPDynamicsintheLeadingLamella IgorR.Kuznetsov*,MarcHerant,MicahDembo BiomedicalEngineering,BostonUniversity,Boston,Massachusetts,UnitedStatesofAmerica Abstract Background:ThetransportoflabeledG-actinfromthemid-lamellaregiontotheleadingedgeinahighlymotilemalignant rat fibroblast line has been studied using fluorescence localization after photobleaching or FLAP, and the transit times recordedintheseexperimentsweresofastthatsimplediffusionwasdeemedaninsufficientexplanation(seeZichaetal., Science,v.300,pp.142–145[1]). Methodology/PrincipalFindings:Were-examinetheZichaFLAPexperimentsusingatwo-phasereactiveinterpenetrating flow formalism to model the cytoplasm and the transport dynamics of bleached and unbleached actin. By allowing an improvedtreatmentofeffectsrelatedtotheretrogradeflowofthecytoskeletonandofthegeometryandfinitethicknessof thelamella,thisnewanalysisrevealsamechanismthatcanrealisticallyexplainthetimingandtheamplitudeofalltheFLAP signalsobservedin[1]withoutinvokingspecialtransportmodalities. Conclusions/Significance:Weconcludethatsimplediffusionissufficenttoexplaintheobservedtransportrates,andthat variationsinthetransportoflabeledactinthroughthelamellaareminorandnotlikelytobethecauseoftheobserved physiologicalvariationsamongdifferentsegmentsoftheleadingedge.Wefindthatsuchvariationsinlabelingcaneasily arise from differences and changes in the microscopic actin dynamics inside the edge compartment, and that the key dynamicalparameterinthisregardistheso-called‘‘dilatationrate’’(thevelocityofcytoskeletalretrogradeflowdividedbya characteristic dimension of the edge compartment where rapid polymerization occurs). If our dilatation hypothesis is correct,thetransientkineticsofbleachedactinrelocalizationconstituteanovelandverysensitivemethodforprobingthe cytoskeletaldynamicsinleadingedgemicro-environmentswhichareotherwiseverydifficulttodirectlyinterrogate. Citation: Kuznetsov IR, Herant M, Dembo M (2010) Analysis of Actin FLAP Dynamics in the Leadi