Delayed Degradation and Impaired Dendritic Delivery of Intron-Lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc Transgenic Mice
dc.contributor.author | Steward, Oswald | en |
dc.contributor.author | Yee, Kelly Matsudaira | en |
dc.contributor.author | Farris, Shannon | en |
dc.contributor.author | Pirbhoy, Patricia S. | en |
dc.contributor.author | Worley, Paul | en |
dc.contributor.author | Okamura, Kohji | en |
dc.contributor.author | Okuno, Hiroyuki | en |
dc.contributor.author | Bito, Haruhiko | en |
dc.date.accessioned | 2019-06-03T21:03:03Z | en |
dc.date.available | 2019-06-03T21:03:03Z | en |
dc.date.issued | 2018-01-31 | en |
dc.description.abstract | Arc 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.sponsorship | Grant 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.extent | 21 pages | en |
dc.format.mimetype | application/pdf | en |
dc.identifier.citation | Steward 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.2017.00435 | en |
dc.identifier.doi | https://doi.org/10.3389/fnmol.2017.00435 | en |
dc.identifier.uri | http://hdl.handle.net/10919/89716 | en |
dc.identifier.volume | 10 | en |
dc.language.iso | en | en |
dc.publisher | Frontiers | en |
dc.rights | Creative Commons Attribution 4.0 International | en |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en |
dc.subject | LTP | en |
dc.subject | synaptic plasticity | en |
dc.subject | protein synthesis | en |
dc.subject | dendrite | en |
dc.subject | dendritic mRNA | en |
dc.subject | dendritic spines | en |
dc.subject | immediate early gene | en |
dc.subject | nonsense-mediated decay | en |
dc.title | Delayed Degradation and Impaired Dendritic Delivery of Intron-Lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc Transgenic Mice | en |
dc.title.serial | Frontiers in Molecular Neuroscience | en |
dc.type | Article - Refereed | en |
dc.type.dcmitype | Text | en |
Files
Original bundle
1 - 1 of 1