Organotypic Brain Slice Cultures of Adult Transgenic P301S Mice—A Model for Tauopathy Studies 英文参考文献.docVIP

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Organotypic Brain Slice Cultures of Adult Transgenic P301S Mice—A Model for Tauopathy Studies 英文参考文献.doc

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Organotypic Brain Slice Cultures of Adult Transgenic P301S Mice—A Model for Tauopathy Studies 英文参考文献

OrganotypicBrainSliceCulturesofAdultTransgenic P301SMice—AModelforTauopathyStudies AgnetaMewes1,HeikeFranke2,DavidSinger1* 1InstituteofBioanalyticalChemistry,CenterforBiotechnologyandBiomedicine(BBZ),UniversityofLeipzig,Leipzig,Germany,2Rudolf-Boehm-InstituteofPharmacology andToxicology,UniversityofLeipzig,Leipzig,Germany Abstract Background:Organotypicbrainsliceculturesrepresentanexcellentcompromisebetweensinglecellculturesandcomplete animalstudies,inthiswayreplacingandreducingthenumberofanimalexperiments.Organotypicbrainslicesarewidely appliedtomodelneuronaldevelopmentandregenerationaswellasneuronalpathologyconcerningstroke,epilepsyand Alzheimer’sdisease(AD).ADischaracterizedbytwoproteinalterations,namelytauhyperphosphorylationandexcessive amyloid b deposition, both causing microglia and astrocyte activation. Deposits of hyperphosphorylated tau, called neurofibrillarytangles(NFTs),surroundedbyactivatedgliaaremodeledintransgenicmice,e.g.thetauopathymodelP301S. Methodology/PrincipalFindings:Inthisstudyweexplorethebenefitsandlimitationsoforganotypicbrainslicecultures madeofmatureadulttransgenicmiceasapotentialmodelsystemforthemultifactorialphenotypeofAD.First,neonatal (P1)andadultorganotypicbrainsliceculturesfrom7-to10-month-oldtransgenicP301Smicehavebeencomparedwith regardtovitality,whichwasmonitoredwiththelactatedehydrogenase(LDH)-andtheMTT(3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) assays over 15 days. Neonatal slices displayed a constant high vitality level, while the vitalityofadultsliceculturesdecreasedsignificantlyuponcultivation.Variouspreparationandcultivationconditionswere tested to augment the vitality of adult slices and improvements were achieved with a reduced slice thickness, a mild hypothermic cultivation temperature and a cultivation CO2 concentration of 5%. Furthermore, we present a substantial immunohistochemical characterization analyzing the morphology of neurons, astrocytes and microglia in comparison to neonataltissue. Con