prdm2 probe  (Advanced Cell Diagnostics Inc)

Journal: Molecular Psychiatry

Article Title: An epigenetic mechanism for over-consolidation of fear memories

doi: 10.1038/s41380-022-01758-6

Figure Lengend Snippet: A Representative tile scan of virus injection site and spread. B , C Representative images used for RNAscope quantification. D KD of Prdm2 induces a significant downregulation of Prdm2 in the dmPFC. E Experimental timeline: On day 1, rats received bilateral infusion of an AAV9 containing either a shRNA- Prdm2 or a scrambled control. Rats were then tested for fear acquisition (day 31), fear expression (day 32), and fear extinction (day 33–34). F KD of Prdm2 did not affect the acquisition of fear memory (acquisition of fear memory is presented as an average of tones 2–6 during the conditioning session), but G significantly increased fear expression 24 h after conditioning (indicated by the % freezing ±SEM; N = 17–20/ group). H Average of the first 2 tones from the fear expression test. I Prdm2 KD did not affect the rate of extinction, indicated by the interaction for time x group not being significant (i.e., similar slopes). J In a separate batch ( N = 12/group), Prdm2 KD was found to increase fear expression also 1w after conditioning. K When Prdm2 was knocked down 1 week after the acquisition of fear memory ( N = 18–20/group), no effect was observed on fear expression measured 1 month later. * p < 0.05; ** p < 0.01; p < 0.001.

Article Snippet: The Prdm2 probe (accession number NM_001077648.1) was purchased from Advanced Cell Diagnostics (Newark, CA, USA).

Techniques: Virus, Injection, shRNA, Expressing

Journal: Molecular Psychiatry

Article Title: An epigenetic mechanism for over-consolidation of fear memories

doi: 10.1038/s41380-022-01758-6

Figure Lengend Snippet: Prdm2 KD does not affect shock sensitivity ( A ), locomotor activity ( B ), novel object recognition ( C ) and basal anxiety ( D ).

Article Snippet: The Prdm2 probe (accession number NM_001077648.1) was purchased from Advanced Cell Diagnostics (Newark, CA, USA).

Techniques: Activity Assay

Journal: Molecular Psychiatry

Article Title: An epigenetic mechanism for over-consolidation of fear memories

doi: 10.1038/s41380-022-01758-6

Figure Lengend Snippet: A Schematic representing the dual viral approach. B Tile scans showing the viral spread in the dmPFC and BLA and representative images showing dmPFC neurons infected by AAV9-DIO-shRNA-PRDM2-ZS green (green), dmPFC neurons infected by rAAV2 retro Cre-mCherry (red) and dmPFC neurons showing co-infection of AAV9-DIO-shRNA-PRDM2-ZS green and rAAV2 retro Cre-mCherry (yellow; scale bar, 50 µm). C KD of Prdm2 in neurons projecting from the dmPFC to the BLA did not affect fear acquisition measured as % freezing ± SEM. D Prdm2 KD in dmPFC-BLA was sufficient to increase fear expression 24 h after conditioning ( N = 19–20). * p < 0.05. BLA basolateral amygdala, dmPFC dorsomedial prefrontal cortex.

Article Snippet: The Prdm2 probe (accession number NM_001077648.1) was purchased from Advanced Cell Diagnostics (Newark, CA, USA).

Techniques: Infection, shRNA, Expressing

Journal: Molecular Psychiatry

Article Title: An epigenetic mechanism for over-consolidation of fear memories

doi: 10.1038/s41380-022-01758-6

Figure Lengend Snippet: Prdm2 KD in dmPFC-BLA neurons regulates genes involved in synaptogenesis. A Schematic representing the triple viral approach used for the vTRAP experiment. B Tile scans showing the viral spread in the dmPFC and BLA as well as dmPFC neurons presenting cells infected by AAV5-FLEX-EGFPL10a (green), cells infected by rAAV2 retro Cre-mCherry (red), and cells showing co-infection of AAV5-FLEX-EGFPL10a and rAAV2 retro Cre-mCherry (yellow). C Principal component analysis showing separation of Prdm2 KD samples and scrambled control into distinct clusters. D Volcano Plot illustrating the most significantly altered genes following Prdm2 KD. E Hierarchical clustering dendrogram grouping together interconnected, highly co-expressed genes. Colormaps beneath the dendrogram corresponds to modules of co-expressed genes. Top colormap: initial identified modules. Bottom colormap: modules after merging modules with similar expression profiles. F Differential expression analysis for each co-expression module, comparing Prdm2 KD with scrambled control. Red horizontal dashed line denotes a significance level of FDR-corrected p value of 0.05. G Boxplot comparing the gene expression profile of module “MEblue” between conditions. KD: Prdm2 KD, SCR: Scramble control. Statistical test: Two-sided unpaired t-test. H Gene ontology enrichment for genes in the module “MEblue”. I Gene network analysis performed using IPA. Prdm2 KD increases expression of genes that code for the cadherin, neurexin/neuroligin and ephrin/ephrin receptors family as well as for proteins that belongs to the SNARE complex. J Differential expression and significance level for selected synaptogenesis-related genes. Vertical dashed line in the bar plot denotes a significance level of FDR-corrected p value of 0.05. BLA basolateral amygdala, dmPFC dorsomedial prefrontal cortex.

Article Snippet: The Prdm2 probe (accession number NM_001077648.1) was purchased from Advanced Cell Diagnostics (Newark, CA, USA).

Techniques: Infection, Expressing

Journal: Molecular Psychiatry

Article Title: An epigenetic mechanism for over-consolidation of fear memories

doi: 10.1038/s41380-022-01758-6

Figure Lengend Snippet: A Representative image showing the viral expression (AAV9.HI.shR.ratPrdm2.CMV.ZsGreen.SV40) in dmPFC terminals targeting the BLA. B Representative sEPSCs traces recorded from BLA putative principal neurons (PNs) in Scrambled or Prdm2 KD rats. Scale bars: 50 pA × 500 ms. Cumulative distributions and bar graphs showing the frequency ( C ) and amplitude ( D ) of sEPSCs recorded from putative BLA PNs in Scrambled or Prdm2 KD rats. E Schematic representation of the recording and stimulating electrodes sites for evoked (e)EPSCs recordings. F Representative eEPSCs traces evoked by paired electrical stimulations recorded from putative BLA PNs in Scrambled or Prdm2 KD rats. Scale bars: 50 pA × 25 ms. G Bar graphs showing the mean values of PPR recorded from putative BLA PNs in Scrambled or Prdm2 KD rats. Bar graphs showing the mean values of 1/ CV 2 ( H ) and VMR ( I ) of eEPSCs recorded from Scrambled or Prdm2 KD rats ( N = 5–6/group). * p < 0.05. J Schematic representing the fiber photometry approach (top) and a representative image (bottom) showing the implanted optical fiber aimed at the BLA and the expression of two viruses, AAV9.HI.shR.ratPrdm2.CMV.ZsGreen.SV40 in dmPFC terminals targeting the BLA as well as AAV9.CaMKII.GCaMP6s.WPRE.SV40 in BLA glutamatergic neurons. K Traces showing normalized calcium-dependent GCaMP fluorescent signal (mean ± SEM) in BLA neurons of Scrambled or Prdm2 KD rats, averaged over the first 2 tones from the fear expression test. Duration of the tone is indicated by the shaded gray box. Bar graphs showing larger peak normalized GCaMP signal in response to tone onset ( L ) and increased Area Under the Curve (AUC) of the normalized GCaMP signal during the 5 s preceding the tone and the first 5 s of the tone presentation ( M ) in Prdm2 KD rats compared to Scrambled control rats. BLA basolateral amygdala, CeA central amygdala, dmPFC dorsomedial prefrontal cortex, LA lateral amygdala, VMR variance to mean ratio, sEPSCs spontaneous excitatory postsynaptic currents.

Article Snippet: The Prdm2 probe (accession number NM_001077648.1) was purchased from Advanced Cell Diagnostics (Newark, CA, USA).

Techniques: Expressing