Organotypic brain slices a model to study the neurovascular unit micro-environment in epilepsies 英文参考文献.docVIP

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Organotypic brain slices a model to study the neurovascular unit micro-environment in epilepsies 英文参考文献.doc

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Organotypic brain slices a model to study the neurovascular unit micro-environment in epilepsies 英文参考文献

Morin-Brureauetal.FluidsandBarriersoftheCNS2013,10:11 /content/10/1/11 FLUIDS AND BARRIERS OF THE CNS STUDY PROTOCOL OpenAccess Organotypicbrainslices:amodeltostudythe neurovascularunitmicro-environmentin epilepsies MélanieMorin-Brureau1,2,FrédéricDeBock1andMireilleLerner-Natoli1* Abstract Background:Itisnowrecognizedthattheneuro-vascularunit(NVU)playsakeyroleinseveralneurological diseasesincludingepilepsy,stroke,Alzheimer’sdisease,multiplesclerosisandthedevelopmentofgliomas.Mostof thesedisordersareassociatedwithNVUdysfunction,duetooverexpressionofinflammatoryfactorssuchas vascularendothelialgrowthfactor(VEGF).Variousinvitromodelshavebeendevelopedpreviouslytostudythe micro-environmentoftheblood–brainbarrier(BBB).Howevernoneoftheseinvitromodelscontainedacomplete complementofNVUcells,normaintainedtheirinteractions,thusminimizingtheinfluenceofthesurrounding tissueontheBBBdevelopmentandfunction.Theorganotypichippocampalculture(OHC)isanintegrativeinvitro modelthatallowsrepeatedmanipulationsovertimetofurtherunderstandthedevelopmentofcellcircuitsorthe mechanismsofbraindiseases. Methods/design:OHCswereculturedfromhippocampiof6–7day-oldSpragueDawleyrats.After2weeksin culture,seizureswereinducedbyapplicationofkainateorbicucullineintoculturemedium.TheregulationofBBB integrityunderphysiologicalandpathologicalconditionswasevaluatedbyimmunostainingofthemaintight junction(TJ)proteinsandofthebasalmembraneofmicrovessels.TomimicorpreventBBBdisassembly,weused diversepro-oranti-angiogenictreatments. Discussion:ThisstudydemonstratesthatNVUregulationcanbeinvestigatedusingOHCs.Weobservedinthis modelsystemanincreaseinvascularizationandadown-regulationofTJproteins,similartothevascularchanges describedinachronicfocusofepilepticpatients,andinrodentmodelsofepilepsyorinflammation.Weobserved thatZonulaoccludens-1(ZO-1)proteindisappearedafterseizuresassociatedwithneuronaldamage.Inthese conditions,theangiopoeitin-1systemwasdown-regulated,andtheapplicationofr-angiopoeitin-1allowedTJ re-assembly.T