Delayed Degradation and Impaired Dendritic Delivery of Intron-Lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc Transgenic Mice

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dc.contributor.authorSteward, Oswalden
dc.contributor.authorYee, Kelly Matsudairaen
dc.contributor.authorFarris, Shannonen
dc.contributor.authorPirbhoy, Patricia S.en
dc.contributor.authorWorley, Paulen
dc.contributor.authorOkamura, Kohjien
dc.contributor.authorOkuno, Hiroyukien
dc.contributor.authorBito, Haruhikoen
dc.date.accessioned2019-06-03T21:03:03Zen
dc.date.available2019-06-03T21:03:03Zen
dc.date.issued2018-01-31en
dc.description.abstractArc is a unique immediate early gene (IEG) whose expression is induced as synapses are modified during learning. Newly-synthesized ArcmRNA is rapidly transported throughout dendrites and localizes near recently activated synapses. Arc mRNA levels are regulated by rapid degradation, which is accelerated by synaptic activity in a translation-dependent process. One possible mechanism is nonsense-mediated mRNA decay (NMD), which depends on the presence of a splice junction in the 3_UTR. Here, we test this hypothesis using transgenic mice that express EGFP-Arc. Because the transgene was constructed from Arc cDNA, it lacks intron structures in the 3_UTR that are present in the endogenous Arc gene. NMD depends on the presence of proteins of the exon junction complex (EJC) downstream of a stop codon, so EGFP-Arc mRNA should not undergo NMD. Assessment of Arc mRNA rundown in the presence of the transcription inhibitor actinomycin-D confirmed delayed degradation of EGFP-Arc mRNA. EGFP-Arc mRNA and protein are expressed at much higher levels in transgenic mice under basal and activated conditions but EGFP-Arc mRNA does not enter dendrites efficiently. In a physiological assay in which cycloheximide (CHX) was infused after induction of Arc by seizures, there were increases in endogenous Arc mRNA levels consistent with translation-dependent Arc mRNA decay but this was not seen with EGFP-Arc mRNA. Taken together, our results indicate: (1) Arc mRNA degradation occurs via a mechanism with characteristics of NMD; (2) rapid dendritic delivery of newly synthesized Arc mRNA after induction may depend in part on prior splicing of the 3_UTR.en
dc.description.sponsorshipGrant support: R01NS12333 to OS, R35NS097966 to PW, JSPS-KAKENHI grants 15H02358 and 17H06312 (to HB) and 15H04258 (to HO). PSP was the recipient of fellowship support from NIH MBRS-IMSD GM055246, NIH 5T32 NS045540, NIH NINDS F31 NS083349.en
dc.format.extent21 pagesen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationSteward O, Matsudaira Yee K, Farris S, Pirbhoy PS, Worley P, Okamura K, Okuno H and Bito H (2018) Delayed Degradation and Impaired Dendritic Delivery of Intron-Lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc Transgenic Mice. Front. Mol. Neurosci. 10:435. doi: 10.3389/fnmol.2017.00435en
dc.identifier.doihttps://doi.org/10.3389/fnmol.2017.00435en
dc.identifier.urihttp://hdl.handle.net/10919/89716en
dc.identifier.volume10en
dc.language.isoenen
dc.publisherFrontiersen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectLTPen
dc.subjectsynaptic plasticityen
dc.subjectprotein synthesisen
dc.subjectdendriteen
dc.subjectdendritic mRNAen
dc.subjectdendritic spinesen
dc.subjectimmediate early geneen
dc.subjectnonsense-mediated decayen
dc.titleDelayed Degradation and Impaired Dendritic Delivery of Intron-Lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc Transgenic Miceen
dc.title.serialFrontiers in Molecular Neuroscienceen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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