MicroRNA Regulation of the Synaptic Plasticity-Related Gene Arc 英文参考文献.docVIP

MicroRNARegulationoftheSynapticPlasticity-Related GeneArc KarinWibrand.,BalagopalPai.,TaweepornSiripornmongcolchai,MargaretheBittins, BirgitteBerentsen,MayLillianOfte,ArwedWeigel,KaiOveSkaftnesmo,CliveR.Bramham* DepartmentofBiomedicineandK.G.JebsenCentreforResearchonNeuropsychiatricDisorders,UniversityofBergen,Bergen,Norway Abstract Expressionofactivity-regulatedcytoskeletonassociatedprotein(Arc)iscrucialfordiversetypesofexperience-dependent synaptic plasticity and long-term memory in mammals. However, the mechanisms governing Arc-specific translation are little understood. Here, we asked whether Arc translation is regulated by microRNAs. Bioinformatic analysis predicted numerouscandidatemiRNAbindingsiteswithintheArc39-untranslatedregion(UTR).Transfectionofthecorresponding microRNAsinhumanembryonickidneycellsinhibitedexpressionofanArc39UTRluciferasereporterfrombetween10to 70%across16microRNAstested.PointmutationanddeletionofthemicroRNA-bindingseed-regionformiR-34a,miR-326, andmiR-19apartiallyorfullyrescuedreporterexpression.Inaddition,expressionofspecificmicroRNApairssynergistically modulatedArcreporterexpression.Inprimaryrathippocampalneuronalcultures,ectopicexpressionofmiR-34a,miR-193a, ormiR-326,downregulatedendogenousArcproteinexpressioninresponsetoBDNFtreatment.Conversely,treatmentof neurons with cell-penetrating, peptide nucleic acid (PNA) inhibitors of miR-326 enhanced Arc mRNA expression. BDNF dramaticallyupregulatedneuronalexpressionofArcmRNAandmiR-132,aknownBDNF-inducedmiRNA,withoutaffecting expressionofArc-targetingmiRNAs.Developmentally,miR-132wasupregulatedatday10invitrowhereasArc-targeting miRNAsweredownregulated.Intheadultbrain,LTPinductioninthedentategyrustriggeredmassiveupregulationofArc and upregulation of miR-132 without affecting levels of mature Arc-targeting miRNAs. Turning to examine miRNA localization, qPCR analysis of dentate gyrus synaptoneurosome and total lysates fractions demonstrated synaptic enrichmentrelativetosmallnucleolarRNA.Inc