Fig. 1: lncRNA D17Rik is enriched in dorsal CA1 of mice after CFC.

A Experimental Design. 1 hr after CFC training brains were fast frozen. The tissue was stained with RNAse-free cresyl violet. The dorsal CA1 was dissected by LCM, processed for RNA isolation followed by RNAseq and DEseq in “R”. B Venn diagrams derived from the DEseq analysis show a higher number of significantly regulated genes in the experimental group (C + S) compared to control groups (context alone and immediate shock) (nominal p-value < 0.05). C Metascape analysis indicates that the majority of genes significantly regulated in C + S condition compared to controls are grouped into a cluster related to early synaptic plasticity. D DEseq results represented by volcano plots show a higher enrichment of the lncRNA D17Rik and plasticity-related genes in the C + S group compared to controls (nominal p-value < 0.05). E Heat map represents log2Foldchange of EGs differentially expression in CFC. F Heat map represents log2 Foldchange of some of the most differentially regulated lncRNAs by C + S in dorsal CA1. G Relative expression of D17rik (Context n = 5, Shock n = 5, C + S n=4 mouse hippocampi) and synaptic plasticity-related genes CamK2b and Egr1 (n = 3 for all conditions) 1 h after training (One-Way ANOVA, Multiple Comparisons Dunnett’s test, D17rik *p = 0.03, CamK2b *p = 0.03, Egr1 *p = 0.04; data is shown as mean ± SEM). H D17Rik is enriched only in CA1 after CFC in the C + S, but not in CA3. There are no significant differences between CA1 and CA3 D17Rik expression in control groups (n = 3-4). Two-way ANOVA + Tukey’s test. **p = 0.002; data is shown as mean ± SEM.) I FISH shows that the lncRNA D17Rik is expressed in mouse hippocampus. High magnification details from a representative mouse’s pyramidal layer in CA1 indicate a mainly cytoplasmic subcellular localization of D17Rik. Photomicrographs show D17Rik(red) and nuclear marker DAPI(blue). Scale bars = 200 µm. This experiment was completed with tissues from 3 mice. Each time produced similar results. J FISH photomicrograph shows that D17Rik is expressed in mouse primary hippocampal neuronal cultures. D17Rik (green) colocalizes with dendritic marker Map2(blue). Cell body scale bar=20 µm. Dendrite inset scale bar=2 µm. This experiment was completed four different times. Each time produced similar results.