Journal: Cell reports
Article Title: Unexpected Transcriptional Programs Contribute to Hippocampal Memory Deficits and Neuronal Stunting after Early-Life Adversity
doi: 10.1016/j.celrep.2020.108511
Figure Lengend Snippet: (A) A schematic of the mechanism by which the NRSE-ODN blocks NRSF function. The sequence of the synthetic, protected oligodeoxynucleotides (ODNs) replicates the recognition site for NRSF (NRSE). The ODNs, flooding the cells, bind available NRSF, acting as decoys and preventing NRSF binding to NRSE on genomic DNA (see also ). (B) H3K9 dimethylation at NRSE sites of the Npas4 gene was increased in adult dorsal hippocampus following ELA. Transiently blocking NRSF binding to chromatin with the use of NRSE-ODNs prevented this long-term ELA-provoked histone modification (significant main effects of ELA, F [1,20] = 6.61, p = 0.02, and of NRSE, F [1,20] = 4.97, p = 0.04; post hoc , p < 0.05). (C) Memory for the prior location of a previously seen object was severely impaired in ELA rats receiving a random ODN sequence (Scrambled [SCR]) but was rescued in those receiving an NRSE-sequence decoy ODN to a level similar to that in control (CTL+SCR) rats. Two-way ANOVA demonstrated a significant interaction of the early-life experience and the type of treatment ( F [1,26] = 16.44, p = 0.0004), and post hoc analyses identified a significant effect of the NRSF blockade intervention (p < 0.05). (D) In an independent measure of spatial memory, ELA+SCR rats could not reliably discriminate between the novel arm and the familiar arm of the Y-maze, which they had explored 4 h earlier (see Y-maze schematic). However, NRSF blockade prevented this spatial memory deficit (significant ELA × NRSE interaction, F [1,25] = 7.26, p = 0.01, post hoc , p < 0.05). Representative heatmaps of the time spent in each area of the Y-maze are shown for each group, with the most time spent indicated with dark red and scaled through the least time spent represented with dark blue. (E) Object recognition memory, which is less hippocampus-dependent, was unaltered by ELA or NRSF manipulation. Data in bar graphs of (C)–(E) are shown as the discrimination index: (time investigating novel location or object – time investigating familiar location or object)/(sum of time investigating both novel and familiar locations or objects). (F–H) In contrast with the rescue of the impaired spatial memory, locomotion (open field test) (F), anxiety-like behaviors (elevated-plus maze) (G) and depressive-like behaviors (Porsolt forced-swim test) (H) were not affected by either the ELA or the ODN-NRSE treatment. Data are presented as mean ± SEM. *p < 0.05. See also .
Article Snippet: Anti-NRSF (H-290) ChIP antibody , Santa Cruz , Cat# sc-25398; RRID: AB_2179625.
Techniques: Sequencing, Binding Assay, Blocking Assay, Modification, Control
Santa Cruz Biotechnology is a verified supplier 
![(A) A schematic of the mechanism by which the NRSE-ODN blocks NRSF function. The sequence of the synthetic, protected oligodeoxynucleotides (ODNs) replicates the recognition site for NRSF (NRSE). The ODNs, flooding the cells, bind available NRSF, acting as decoys and preventing NRSF binding to NRSE on genomic DNA (see also ). (B) H3K9 dimethylation at NRSE sites of the Npas4 gene was increased in adult dorsal hippocampus following ELA. Transiently blocking NRSF binding to chromatin with the use of NRSE-ODNs prevented this long-term ELA-provoked histone modification (significant main effects of ELA, F [1,20] = 6.61, p = 0.02, and of NRSE, F [1,20] = 4.97, p = 0.04; post hoc , p < 0.05). (C) Memory for the prior location of a previously seen object was severely impaired in ELA rats receiving a random ODN sequence (Scrambled [SCR]) but was rescued in those receiving an NRSE-sequence decoy ODN to a level similar to that in control (CTL+SCR) rats. Two-way ANOVA demonstrated a significant interaction of the early-life experience and the type of treatment ( F [1,26] = 16.44, p = 0.0004), and post hoc analyses identified a significant effect of the NRSF blockade intervention (p < 0.05). (D) In an independent measure of spatial memory, ELA+SCR rats could not reliably discriminate between the novel arm and the familiar arm of the Y-maze, which they had explored 4 h earlier (see Y-maze schematic). However, NRSF blockade prevented this spatial memory deficit (significant ELA × NRSE interaction, F [1,25] = 7.26, p = 0.01, post hoc , p < 0.05). Representative heatmaps of the time spent in each area of the Y-maze are shown for each group, with the most time spent indicated with dark red and scaled through the least time spent represented with dark blue. (E) Object recognition memory, which is less hippocampus-dependent, was unaltered by ELA or NRSF manipulation. Data in bar graphs of (C)–(E) are shown as the discrimination index: (time investigating novel location or object – time investigating familiar location or object)/(sum of time investigating both novel and familiar locations or objects). (F–H) In contrast with the rescue of the impaired spatial memory, locomotion (open field test) (F), anxiety-like behaviors (elevated-plus maze) (G) and depressive-like behaviors (Porsolt forced-swim test) (H) were not affected by either the ELA or the ODN-NRSE treatment. Data are presented as mean ± SEM. *p < 0.05. See also .](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7243/pmc07817243/pmc07817243__nihms-1657012-f0005.jpg)
![(A) Sholl analyses of dorsal hippocampal pyramidal neurons in area CA1 of adult rats revealed that ELA stunted dendritic arborization (significant ELA × distance from soma interaction, F [38,494] = 1.90, p = 0.0013; post hoc , p < 0.05). Interfering with NRSF binding rescued neuronal dendritic complexity of ELA rats to control levels (significant NRSE × distance from soma interaction, F [38,494] = 2.46, p < 0.0001; post hoc , p < 0.05). (B) A similar pattern of diminished dendritic complexity was observed in area CA3 of ELA rats (significant ELA × distance from soma interaction, F [54,702] = 1.60, p = 0.005; post hoc , p < 0.05). (C) In the dentate gyrus, NRSE ODNs overall enhanced dendritic complexity (significant NRSE × distance from soma interaction, F [40,520] = 2.85, p < 0.0001), with no significant influence of ELA. (D) Treatment of developing hippocampal neurons in vitro with NRSE ODNs, but not SCR ODNs, markedly increased dendritic complexity in pyramidal-like neurons (short/long axis ratio < 1.5; significant NRSE × distance from soma interaction, F [118,1593] = 1.79, p < 0.0001; post hoc , p < 0.05). (E) NRSE ODNs labeled with BODIPY are observed inside the nuclei of cultured hippocampal neurons 40 h after a 2-h incubation with the ODNs, confirming that they are readily able to enter the cell and concentrate in the nucleus. Scale bar: 10 μm. See also .](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7243/pmc07817243/pmc07817243__nihms-1657012-f0006.jpg)
