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Abstract
BC1 RNA is a small brain specific non-protein coding RNA. It is transported from the cell body into dendrites where it is involved in the fine-tuning translational control. Due to its compactness and established secondary structure, BC1 RNA is an ideal model for investigating the motifs necessary for dendritic localization. Previously, microinjection of in vitro transcribed BC1 RNA mutants into the soma of cultured primary neurons suggested the importance of RNA motifs for dendritic targeting. These ex vivo experiments identified a single bulged nucleotide (U22) and a putative K-turn (GA motif) structure required for dendritic localization or distal transport, respectively. We generated six transgenic mouse lines (three founders each) containing neuronally expressing BC1 RNA variants on a BC1 RNA knockout mouse background. In contrast to ex vivo data, we did not find indications of reduction or abolition of dendritic BC1 RNA localization in the mutants devoid of the GA motif or the bulged nucleotide. We confirmed the ex vivo data, which showed that the triloop terminal sequence had no consequence on dendritic transport. Interestingly, changing the triloop supporting structure completely abolished dendritic localization of BC1 RNA. We propose a novel RNA motif important for dendritic transport in vivo.
Highlights
BC1 RNA gene (BC1−/−)knockout mouse line was chosen as recipient background[11]
Much is known regarding the movement of RNA between cellular compartments; this includes the transport of selected messenger RNAs (mRNAs) and various other RNAs, mostly those involved in translation and its regulation from the cell bodies of neurons to their dendritic processes[38]
There were conflicting ex and in vivo data concerning the ability of the 5′stem of BC1 RNA to convey transport competence to chimeric mRNAs
Summary
Our ISH experiments with TgBC1_AUC mice revealed that mutated BC1 RNA distribution in dendrites was not markedly different from wild-type RNA (Fig. 4C,D). We examined a BC1 RNA variant, in which the internal loop nucleotides AAG48–51 had been replaced by UCU48–51 in order to remove the GA motif by generating standard Watson Crick base pairing, extending the canonical RNA alpha helix (Fig. 5B). When ISH was performed on brain slides of TgBC1_GAU mice we did not observe a reduction in the dendritic transport of mutated BC1 RNA in comparison to wild-type animals (Fig. 6C,D). This is in agreement with ex vivo experiments[18]. The signal in the hippocampal CA2 dendritic field indicates that this variant of BC1 RNA is transported to the distal extensions of dendrites
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