简介：Object:ToidentifytranscriptvariantsandexpressionpatternsofporcineMitf.Materialsandmethods:ApairwiseBLASTsearchatNCBIdatabasewasperformedtodeducethestructureofporcineMitfgene.Subsequently,50RACEandfluorescentquantitativeRT-PCRwereusedtoanalyzetheexpressionpatternofporcineMitfindifferenttissues.Results:FourtranscriptvariantsofporcineMitf,MITF-A,MITF-H,MITF-MandMITF-SUSwereidentified,allsharinghighhomologywiththoseinhumans,exceptMitf-SUS.Conclusion:ThesequenceofporcineMitfappearhighlyhomologoustohumanMITF.However,only4transcriptvariantsofporcineMitfwereidentifiedintheseminipigs,lessthanthe9transcriptvariantsinhumanMITF.

简介：ObjectiveTocharacterizemicroRNA(miRNA)expressionprofileinmicrodissectedauditoryepitheliafromtheCorti'sOrganinnewbornandadultrats.MethodsTheTaqManMicroRNAArrayswereusedtoidentifyexpressionofmicroRNAinthenewbornandadultgroups.GOanalysiswasappliedtoanalyzethemainfunctionofthedifferentialexpressiongenesaccordingtotheGeneOntologywhichisthekeyfunctionalclassificationofNCBI.Similarly,PathwayanalysiswasusedtofindoutthesignificantpathwayofthedifferentialgenesaccordingtoKEGG,BiocartaandReatome.ResultsIncreasedexpressionwasseenin16miRNAsinmatureratcomparedtonewbornrats,withincreasedfoldingrangingfrom17to600folds.Expressionlevelsin2miRNAswerereducedinmaturerats,namelyrno-miR-29candrno-miR-29a.Thehigh-enrichmentGOstargetedbyover-expressedmiRNAswerenegativeregulationofepithelialcelldifferentiation,common-partnerSMADproteinphosphorylation,mesenchymal-epithelialcellsignaling,regulationoftransforminggrowthfactorbeta2production,etc.FunctionalanalysisofmiRNAsbyKEGGrevealedthat19signaltransductionpathwayswereupregulatedand14weredownregulated.ConclusionsThedifferenceinmiRNAexpressionpatternsintheorganofCortibetweenneonatalandadultratsmaybecloselyrelatedtomaturationoftheorganofCortiandlossofproliferativecapacityofinnerearhaircells,andTGFβsignalingmayplayanimportantroleinhaircellsregeneration.

简介：Cisplatindamagescochlearhaircellsandspiralganglionneuronsthroughcelldeathsignalingpathwaysthatarenotfullyunderstood.Weusedfocusedapoptosisgenemicroarraystostudyearlychangesingeneexpres-sionincochlearculturesfromP3neonatalratstreatedwithcisplatin(0.2mM).After12hoursofcisplatintreat-ment,morethan50%ofthe96genesonthearrayshowedasignificantdecreaseinexpression,consistentwithwidespreadcelldeath.However,after3hoursofcisplatintreatment,10genesshowedsignificantincreaseinex-pressionintotalcochleartissue.Inexperimentswithsubsetsofcochleartissues,at3h,cisplatininducedincreasedexpressionof12genesinthecochlearsensoryepithelium(basilarmembrane)and11genesinthespiralganglion(tissueofRosenthal'scanal,containingthespiralganglion).Theseincludedpro-andanti-apoptoticgenesin-volvedinthep53signalingpathway,TNFreceptorfamily,NF-kappaBpathway,deathdomainfamily,deatheffec-tordomainfamily,Bcl-2family,CARDfamily,TRAFfamily,andGTPsignaltransduction.Althoughthechangesingeneexpressionshowedanoverlapbetweenbasilarmembraneandspiralganglion,otherchanges,whichmayreflecttheuniqueresponseofeachtissue,werealsoobserved.Pifithrin-αblockedcisplatin-inducedup-regulationofgenesinthep53signalingpathwaywhenassayedbybothsuperarrayandrealtimePCR.Thedataaddtoourunderstandingoftheinvolvementofp53incisplatin-inducedototoxicityandotoprotection,conferredbythep53inhibitorPifithrin-α.

简介：NuclearfactorkappaB(NF-κB)isoneofthebest-characterizedtranscriptionfactorsplayingimportantrolesinmanycellularresponsestoalargevarietyofstimuli,includinginflammatorycytokines,phorbolesters,growthfactors,andbacterialandviralproducts.TheaimofthisstudyistodemonstrateNF-κBexpressioninthemousecochleaanditsenhancementinresponsetolipopolysaccharides(LPS)andkanamycin(KA)treatment.MethodsKAtreatmentconsistedofsubcutaneousKAinjectionsat700mg/kgtwiceadaywithaneight-hourintervalbetweenthetwoinjectionsfor3or7days.ForanimalsintheLPStreatmentgroup,asingledoseof0.3mgLPSdissolvedin0.2mlsterilesalinewereinjectedintobothbullaethroughthetympanicmembraneandkepttherefor3hours.Animalsinthecontrolgroupreceivedsubcutaneoussalineinjectionfor7days.Followingimmmunohistochemichalprocessingwithrabbitpolyclonalanti-NF-κBp65antibodies,cryosectionsofthecochleawereexaminedforexpressionofNF-κBp65invariousstructuresinthecochlea.ResultsNF-κBp65expression,identifiedbypresenceofbrownreactionproductscharacteristicofDABimmunohistochemistry,wasvisibleinthespiralligament,spiralprominence,tectorialmembrane(TM),spiralganglionandnervefibers.RelativelyweakNF-κBp65expressionwasalsovisualizedintheorganofCorti.WithintheorganofCorti,theinnerhaircells(IHC),outerhaircells(OHC),innerpillarcells(IP),outerpillarcells(OP),Deiter'scells(DC),andBoettcher'scellsexhibitedstrongerstainingthantheinnersulcuscells,Hensen'scells(HC)andClaudius'cells.NoNF-κBp65expressionwasseeninthenucleusoftheIHCandOHC.NF-κBp65expressionwasincreasedinanimalsexposedtoLPSorKA,demonstratingsignificantdifferencesinthestainingbetweencontrolanimalsandLPS/KA-treatedanimals.NF-κBp65expressionwasnotsignificantlydifferentbetweenLPStreatedandKAtreatedanimalsorbetween3and7daysinKA-treatedanimals.Conclusio

简介：ObjectiveTostudytheeffectofsalicylateontheexpressionandfunctionofNMDAreceptorsinspiralganglionneurons(SGNs).MethodsThemRNAofNR1subunitofNMDAreceptorinmodiolustissuesweredetectedbyRealtimefluorescencequantitativePCR(FQ-PCR).NMDAreceptorwhole-cellcurrentswererecordedusingpatchclampinacuteisolatedSGNs.ResultsComparedwiththecontrolgroup,salicylatesignificantlyincreasedthemRNAlevelofNR1subunitinSGNs.NMDAofconcentrationsrangingfrom0.1mMto10mMevokednocurrentinSGNs.NMDA(0.1mMand0.5mM)appliedwithsalicylate(5mM),however,inducedinwardcurrents(212.6±15.2pA,n=5;607.9±44.3pA,n=5)inadose-dependentmanner,whichcouldbeinhibitedbyAPV.SalicylatealonedidnotproduceanycurrentinSGNs.ConclusionSalicylateincreasestheexpressionofNMDAreceptorsandfacilitatesthecurrentsmediatedbyNMDAreceptorsinSGNs.

简介：ObjectiveToconstructaprokaryoticexpressionvectorbearingfusiongeneNT4-ADNF-9forfuturestudiesongenetictherapiesforsensorineuraldeafness.MethodsDoublestrandADNF-9cDNAwassynthesizedusingasymmetricalprimer/templatesandligatedtothe3'terminalofsignalandleaderpeptidesofneurotrophin4(NT4).ThefusiongeneNT4-ADNF-9,wassubclonedintoprokaryoticexpressionvectorpBV220,andnamedpBV220/NT4-ADNF-9.DNAsequenceofthefusiongenewasanalyzed.ThefusionproteinwasisolatedbySDS-PAGEanditsbioactivitywasevaluatedusingprimarycultureofday8chickenembryonicDRGcells.ResultsThecorrectsequenceoffusiongeneNT4-ADNF-9wassuccessfullysubclonedintothepBV220vector.TheexpressedADNF-9proteinshoweditseffectsinpromotingcellsurvivalandneuritegrowth.ConclusionProkaryoticexpressionvectorpBV220/NT4-ADNF-9wasconstructedsuccessfullyandtheexpressedfusionproteindemonstratedsatisfactorybioactivity.

简介：miRNA-183family,innormalbiology,isexpressedinaharmoniousandstablemannerintheneurosensoryorgansandcells.StudieshavealsoshownthatmiRNA-183family,indifferentpathways,affectstheneurosensorydevelopment,maintenance,survivalandfunction.Inaddition,ithaspotentialneuroprotectiveeffectsinresponsetoneurosensorydestructivestimulations.miRNA-96mutationcauseshereditarydeafnessinhumansandmice,andthereforeaffectstheinnerearactivityanditsmaintenance.CertainroleshavebeenidentifiedformiR-96inthemaintenanceandfunctionoftheinnerear.Thecomparisonofthetargetgenesoffamily-183intranscriptomesofnewbornandadulthaircellsshowsthathundredsoftargetgenesinthisfamilymayaffectdevelopmentandmaintenanceoftheears.IdentifyingthegenesthatareregulatedbymiRNA-183familyprovidesresearcherswithimportantinformationaboutthecomplexdevelopmentandenvironmentalregulationoftheinnerear,andcanoffernewapproachestothemaintenanceandregenerationofhaircellsandauditorynerve.

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简介：Thedevelopmentandplasticityofcentralauditorysystemcanbeinfluencedbythechangeofperipheralneuronalactivity.However,themolecularmechanismparticipatingintheprocessremainselusive.Brain-derivedneurotrophicfactor(BDNF)bindingwithitsfunctionalreceptortropomyosinreceptorkinaseB(TrkB)hasmultipleeffectsonneurons.Hereweusedaratmodelofauditorydeprivationbybilateralcochlearablation,toinvestigatethechangesinexpressionofBDNFandTrkBintheauditorycortexafterauditorydeprivationthatoccurredduringthecriticalperiodforthedevelopmentofcentralauditorysystem.Reversetranscription-quantitativepolymerasechainreaction(RTqPCR)andimmunohistochemistrymethodswereadoptedtodetectthemRNAandproteinexpressionlevelsofBDNFandTrkBintheauditorycortexat2,4,6and8weeksaftersurgery,respectively.ThechangeintheexpressionofBDNFandTrkBmRNAsandproteinsfollowedsimilartrend.Inthebilateralcochlearablationgroups,theBDNF-TrkBexpressionlevelinitiallydecreasedat2weeksbutincreasedat4weeksfollowedbythereductionat6and8weeksaftercochlearremoval,ascomparedtotheage-matchedshamcontrolgroups.Inconclusion,theBDNF-TrkBsignalingisinvolvedintheplasticityofauditorycortexinanactivity-dependentmanner.

简介：Noise-inducedhearinglossisacommoncauseofacquiredhearinglossintheadultpopulation.Acousticoverstimulationcausescochleardamagethroughmechanicalstresstothetissue.Consequently,complexmolec-ularchangesareinitiated,andthesechangesleadtomorphologicalandbiologicalalterationsinthecochlea,whichinturncompromisethecochlearfunctionandcausehearingloss.Inthepast10years,therehavebeensignificantadvancesinourunderstandingofthemolecularmechanismsofnoise-inducedhearingloss.Theseadvancesareattributed,inpart,tothedevelopmentofhigh-throughputtechnologiesfortheglobalanalysesofmolecularchanges.Inthisreview,webrieflydescribethenewlydevelopedmethodsforinvestigatingthemo-lecularresponsesofthecochleatoacoustictraumaandtheknowledgegeneratedfromthesestudies.Wealsodiscussthestrengthsandlimitationsofeachtechniqueandthemajorchallengestoinvestigatecochleardegen-erationfollowingacousticinjury.