MeCP2 Dependent Heterochromatin Reorganization during Neural Differentiation of a Novel Mecp2-Deficient Embryonic Stem Cell Reporter Line 英文参考文献.docVIP

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MeCP2 Dependent Heterochromatin Reorganization during Neural Differentiation of a Novel Mecp2-Deficient Embryonic Stem Cell Reporter Line 英文参考文献.doc

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MeCP2 Dependent Heterochromatin Reorganization during Neural Differentiation of a Novel Mecp2-Deficient Embryonic Stem Cell Reporter Line 英文参考文献

MeCP2DependentHeterochromatinReorganization duringNeuralDifferentiationofaNovelMecp2-Deficient EmbryonicStemCellReporterLine BiancaBertulat1.,MariaLuigiaDeBonis2.,FlorianaDellaRagione2,3.,AnneLehmkuhl1, ManuelaMilden1,ChristianStorm1,K.LaurenceJost1,SimonaScala3,BrianHendrich4, MaurizioD’Esposito2,3*,M.CristinaCardoso1* 1DepartmentofBiology,TechnischeUniversita¨t Darmstadt,Darmstadt,Germany,2InstituteofGeneticsandBiophysics‘‘ABuzzatiTraverso’’,Naples,Italy,3Istitutodi Ricovero e Cura a Carattere Scientifico Neuromed, Pozzilli, Italy, 4Wellcome Trust - Medical Research Council Stem Cell Institute and Department of Biochemistry, UniversityofCambridge,Cambridge,UnitedKingdom Abstract TheX-linkedMecp2isaknowninterpreterofepigeneticinformationandmutatedinRettsyndrome,acomplexneurological disease. MeCP2 recruits HDAC complexes to chromatin thereby modulating gene expression and, importantly regulates higher order heterochromatin structure. To address the effects of MeCP2 deficiency on heterochromatin organization duringneuraldifferentiation,wedevelopedaversatilemodelforstemcellinvitrodifferentiation.Therefore,wemodified murineMecp2deficient(Mecp22/y)embryonicstemcellstogeneratecellsexhibitinggreenfluorescentproteinexpression upon neural differentiation. Subsequently, we quantitatively analyzed heterochromatin organization during neural differentiationinwildtypeandinMecp2deficientcells.WefoundthatMeCP2proteinlevelsincreasesignificantlyduring neural differentiation and accumulate at constitutive heterochromatin. Statistical analysis of Mecp2 wild type neurons revealedasignificantclusteringofheterochromatinpernucleiwithprogressingdifferentiation.IncontrastwefoundMecp2 deficientneuronsandastrogliacellstobesignificantlyimpairedinheterochromatinreorganization.Ourresults(i)introduce a new and manageable cellular model to study the molecular effects of Mecp2 deficiency, and (ii) support the view of MeCP2 as a central protein in heterochromatin architecture in maturating cell