Overlapping profiles of Aβ peptides in the Alzheimers disease and pathological aging brains 英文参考文献.docVIP

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Overlapping profiles of Aβ peptides in the Alzheimers disease and pathological aging brains 英文参考文献.doc

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Overlapping profiles of Aβ peptides in the Alzheimers disease and pathological aging brains 英文参考文献

Mooreetal.Alzheimer’sResearchTherapy2012,4:18 /content/4/3/18 RESEARCH ARTICLE OpenAccess OverlappingprofilesofAbpeptidesinthe Alzheimer’sdiseaseandpathologicalagingbrains BrendaDMoore1,ParamitaChakrabarty1,YonaLevites1,TomLKukar2,Ann-MarieBaine3,TinaMoroni3, ThomasBLadd1,PritamDas3,DennisWDickson3andToddEGolde1* Abstract Introduction:AhallmarkofAlzheimer’sdisease(AD)isthepresenceofsenileplaquescomposedofaggregated amyloidb(Ab)peptides.Pathologicalaging(PA)isapostmortemclassificationthathasbeenusedtodescribe brainswithplaquepathologysimilarinextenttoAD,minimalcorticaltaupathology,andnoaccompanyinghistory ofcognitivedeclineinthebraindonorpriortodeath.PAmayrepresenteitheraprodromalphaseofAD,abenign formofAbaccumulation,orinherentindividualresistancetothetoxiceffectsofAbaccumulation.Toattemptto distinguishbetweenthesepossibilitieswehavesystematicallycharacterizedAbpeptidesinapostmortemseriesof PA,ADandnon-dementedcontrol(NDC)brains. Methods:Abwassequentiallyextractedwithtrisbufferedsaline(TBS),radioimmunoprecipitationbuffer(RIPA),2% sodiumdodecylsulfate(SDS)and70%formicacid(FA)fromthepre-frontalcortexof16AD,eightPA,andsixNDC patients.Theseextractswereanalyzedby1)apanelofAbsandwichELISAs,2)immunoprecipitationfollowedby massspectrometry(IP/MS)and3)westernblotting.ThesestudiesenabledustoassesAblevelsandsolubility, peptideprofilesandoligomericassemblies. Results:Inalmostallextracts(TBS,RIPA,2%SDSand70%FA)theaveragelevelsofAb1-40,Ab1-42,Abtotal,and Abx-42weregreatestinAD.Onaverage,levelswereslightlylowerinPA,andtherewasextensiveoverlapbetween AblevelsinindividualPAandADcases.TheprofilesofAbpeptidesdetectedusingIP/MStechniquesalsoshowed extensivesimilaritybetweenthePAandADbrainextracts.InselectADbrainextracts,wedetectedmoreamino- terminallytruncatedAbpeptidescomparedtoPApatients,butthesepeptidesrepresentedaminorportionofthe Abobserved.NoconsistentdifferencesintheAbassemblieswereobservedbywesternblottinginthePAandAD groups. Conclusions:Wefoundextensiveoverlapwithonlysubtle