Regulation of Emx2 Expression by Antisense Transcripts in Murine Cortico-Cerebral Precursors 英文参考文献.docVIP

RegulationofEmx2ExpressionbyAntisenseTranscripts inMurineCortico-CerebralPrecursors GiuliaSpigoni,ChiaraGedressi,AntonelloMallamaci* InternationalSchoolforAdvancedStudies(SISSA/ISAS),Trieste,Italy Abstract Background: Emx2 encodes for a transcription factor expressed in the embryonic intermediate mesoderm and central nervous system (CNS). It is implicated in several aspects of cerebral cortex development, including morphogenetic field specification, arealization, precursor proliferation and lamination. Four Emx2-associated antisense transcripts have been found in the urogenital system; one of them, Emx2OS, has been also detected in the adult brain. Until now, however, nothingisknownaboutexpressionandfunctionofEmx2OSinthedevelopingCNS. Methodology/Principal Findings: By quantitative RT-PCR and in situ hybridization, we reconstructed the Emx2OS expressionprofileintheembryonicCNS,payingspecialattentiontothedevelopingcerebralcortex.Emx2OSwasobserved inanumberofCNSstructuresexpressingalsoEmx2.Withinthecortex,Emx2OSwasdetectableinperiventricularprecursors, expressing the sense transcript, and peaked in newly born post-mitotic neurons not expressing such transcript. By integrating lentiviral gene delivery, RNAi, TetON technology, morpholino-mediated gene knock-down, drug-induced perturbation of gene expression, and quantitative RT-PCR, we addressed possible roles of Ex2 antisense RNA in Emx2 regulation,inprimaryCNSprecursorcultures.Wefoundthat,inbothcorticalprecursorsandtheirneuronalprogenies,Emx2 antisense RNA contributes to post-transcriptional down-regulation of its sense partner, possibly by a Dicer-promoted mechanism.ThesameRNA,whendeliveredtorhombo-spinalprecursors,stimulatesectopicexpressionofEmx2 ,whereas Emx2 knock-out dramatically impairs Emx2OS transcription. This suggests that, within the developing CNS, a reciprocal Emx2/Emx2OSregulatoryloopmaynormallysustaintranscriptionattheEmx2locus. Conclusions/Significance:Thisstudyshowsthatantisensetranscriptsmaycont