Journal: bioRxiv

Article Title: Reduced Prefrontal PRDM2 Increases Stress-Induced Reinstatement Across Sexes via a dmPFC-Nucleus Accumbens Pathway

doi: 10.64898/2026.03.12.711276

Figure Lengend Snippet: (A) Postmortem human prefrontal cortex (PFC) tissue was collected and analyzed to measure PRDM2 expression using quantitative real-time PCR. (B) Experimental design in rats. Male and female rats were trained for alcohol self-administration followed by stereotaxic viral injections targeting the dorsomedial prefrontal cortex (dmPFC) or dmPFC neurons projecting to the nucleus accumbens (NAc) core to induce Prdm2 knockdown (KD) or scrambled control expression. After recovery, rats underwent alcohol self-administration training (fixed-ratio schedule) followed by extinction sessions. Stress-induced reinstatement of alcohol seeking was then tested using intermittent footshock stress at increasing intensities (0.4, 0.6, and 0.8 mA) across separate test sessions. Behavioral responding during reinstatement was compared with the preceding extinction baseline. After completion of behavioral testing, brain tissue was collected for verification of viral expression and molecular analyses.

Article Snippet: Inventoried TaqMan Gene expression assay probes (PRDM2: Hs00202013_m1 and ACTB : Hs01060665_g1; Life Technologies, CA, USA) and TaqManTM Fast Universal PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania) were used to assess gene expression on a QuantStudio 7 Flex Real-time PCR system (Life Technologies, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Knockdown, Control

Journal: bioRxiv

Article Title: Reduced Prefrontal PRDM2 Increases Stress-Induced Reinstatement Across Sexes via a dmPFC-Nucleus Accumbens Pathway

doi: 10.64898/2026.03.12.711276

Figure Lengend Snippet: Relative PRDM2 levels in the prefrontal cortex of male (blue) and female (red) control subjects and individuals with alcohol use disorder (AUD). PRDM2 expression was significantly reduced in AUD compared with controls (***p < 0.001). In addition, a significant sex effect was observed, with females showing lower PRDM2 levels compared with males (###p < 0.001). Bars represent mean ± SEM.

Article Snippet: Inventoried TaqMan Gene expression assay probes (PRDM2: Hs00202013_m1 and ACTB : Hs01060665_g1; Life Technologies, CA, USA) and TaqManTM Fast Universal PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania) were used to assess gene expression on a QuantStudio 7 Flex Real-time PCR system (Life Technologies, CA, USA).

Techniques: Control, Expressing

Journal: bioRxiv

Article Title: Reduced Prefrontal PRDM2 Increases Stress-Induced Reinstatement Across Sexes via a dmPFC-Nucleus Accumbens Pathway

doi: 10.64898/2026.03.12.711276

Figure Lengend Snippet: (A) Representative images showing viral targeting of the dorsomedial prefrontal cortex (dmPFC). Top: schematic showing viral expression restricted to the dmPFC. Bottom: higher magnification image of AAV-shPrdm2 expression (mScarlet, red) with DAPI nuclear staining (blue). (B) Validation of Prdm2 knockdown in the dmPFC using RNAscope in situ hybridization. Representative images from scrambled control and Prdm2 KD animals and quantification of Prdm2 mRNA puncta per cell demonstrating reduced expression following knockdown. (C–E) Stress-induced reinstatement of alcohol seeking in male rats following intermittent footshock at 0.4 mA (C), 0.6 mA (D), and 0.8 mA (E). Stress exposure increased lever pressing relative to extinction conditions across shock intensities, with Prdm2 KD significantly enhancing reinstatement responding at 0.6 mA. (F–H) Stress-induced reinstatement of alcohol seeking in female rats tested at 0.4 mA (F), 0.6 mA (G), and 0.8 mA (H). Similar to males, Prdm2 KD significantly increased reinstatement responding at 0.6 mA but not at the lower or higher shock intensities. Bars represent mean ± SEM lever presses during the 30-min session. *p < 0.05, **p < 0.01; ###p < 0.001 main effect of stress compared with extinction.

Article Snippet: Inventoried TaqMan Gene expression assay probes (PRDM2: Hs00202013_m1 and ACTB : Hs01060665_g1; Life Technologies, CA, USA) and TaqManTM Fast Universal PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania) were used to assess gene expression on a QuantStudio 7 Flex Real-time PCR system (Life Technologies, CA, USA).

Techniques: Expressing, Staining, Biomarker Discovery, Knockdown, RNAscope, In Situ Hybridization, Control

Journal: bioRxiv

Article Title: Reduced Prefrontal PRDM2 Increases Stress-Induced Reinstatement Across Sexes via a dmPFC-Nucleus Accumbens Pathway

doi: 10.64898/2026.03.12.711276

Figure Lengend Snippet: (A) Footshock sensitivity thresholds measured as the current intensity required to elicit paw withdrawal responses (1, 2, or 4 paws retracted) in scrambled and Prdm2 KD female rats. No differences in shock sensitivity were observed between groups, indicating that altered reinstatement behavior is not due to changes in pain perception. (B–D) Effect of estrous cycle stage on stress-induced reinstatement of alcohol seeking following intermittent footshock at 0.4 mA (B), 0.6 mA (C), and 0.8 mA (D). Lever presses during the 30-min reinstatement test are shown for female rats tested during proestrus/estrus (P) and non-proestrus/non-estrus (Non-P) stages of the cycle. Estrous stage did not influence reinstatement responding and did not interact with group at any shock intensity, including 0.6 mA, where Prdm2 KD enhanced reinstatement. Bars represent mean ± SEM. Numbers within bars indicate sample sizes.

Article Snippet: Inventoried TaqMan Gene expression assay probes (PRDM2: Hs00202013_m1 and ACTB : Hs01060665_g1; Life Technologies, CA, USA) and TaqManTM Fast Universal PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania) were used to assess gene expression on a QuantStudio 7 Flex Real-time PCR system (Life Technologies, CA, USA).

Techniques:

Journal: bioRxiv

Article Title: Reduced Prefrontal PRDM2 Increases Stress-Induced Reinstatement Across Sexes via a dmPFC-Nucleus Accumbens Pathway

doi: 10.64898/2026.03.12.711276

Figure Lengend Snippet: PRDM2 in dmPFC → NAc projection neurons regulates stress-induced reinstatement of alcohol seeking in male and female rats. (A–C) Stress-induced reinstatement of alcohol seeking in male rats following intermittent footshock at 0.4 mA (A), 0.6 mA (B), and 0.8 mA (C). Stress exposure significantly increased lever pressing compared with extinction conditions across shock intensities. Prdm2 knockdown in dmPFC → NAc neurons significantly enhanced reinstatement responding at 0.4 mA, but not at 0.6 or 0.8 mA. (D) Footshock sensitivity thresholds in male rats measured as the current intensity required to elicit increasing paw withdrawal responses (1, 2, or 4 paws retracted). No differences were observed between scrambled and Prdm2 KD groups. (E–F) Stress-induced reinstatement of alcohol seeking in female rats tested at 0.4 mA (E), 0.6 mA (F), and 0.8 mA (G). Prdm2 knockdown in dmPFC → NAc neurons significantly increased reinstatement responding at 0.6 mA, but not at 0.4 or 0.8 mA. (H) Footshock sensitivity thresholds in female rats. Prdm2 knockdown did not alter pain sensitivity to footshock. Bars represent mean ± SEM lever presses during the 30-min session. *p < 0.05; ##p < 0.01, ###p < 0.001 main effect of stress compared with extinction.

Article Snippet: Inventoried TaqMan Gene expression assay probes (PRDM2: Hs00202013_m1 and ACTB : Hs01060665_g1; Life Technologies, CA, USA) and TaqManTM Fast Universal PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania) were used to assess gene expression on a QuantStudio 7 Flex Real-time PCR system (Life Technologies, CA, USA).

Techniques: Knockdown

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Syringetin relieves bone cancer pain in rats induced by breast cancer cells through the ESR1/PRDM2 axis

doi: 10.4196/kjpp.24.303

Figure Lengend Snippet: (A) Cell viability was detected by CCK8 after SHZ-88 cells were intervened with 2.5 μM, 5 μM, and 10 μM Syringetin. (B) TUNEL staining was employed to detect the cell apoptosis under the intervention of 10 μM Syringetin (200×). (C) Transwell was applied to detect cell migration under the intervention of 10 μM Syringetin. (D) WB detection of the expression of ESR1 and PRDM2 proteins in cells under the intervention of 10 μM Syringetin. N = 3 replicates/group. ESR1, estrogen receptor 1; CCK8, Cell-Counting-Kit-8; TUNEL, Terminal transferase dUTP Nick End Labeling; WB, Western blotting; DAPI, 4,6-diamidine-2-phenylindole dihydrochloride; PRDM2, positive regulatory domain zinc finger protein2. *p < 0.05 vs . Control.

Article Snippet: Then, first antibodies were added onto the membrane, including ESR1 (21244-1-AP, 1:2,000, Proteintech), PRDM2 (27718-1-AP, 1:500, Proteintech) and β-actin ( AWA80002 , 1:5,000, Abiowell).

Techniques: TUNEL Assay, Staining, Migration, Expressing, Cell Counting, End Labeling, Western Blot, Control

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Syringetin relieves bone cancer pain in rats induced by breast cancer cells through the ESR1/PRDM2 axis

doi: 10.4196/kjpp.24.303

Figure Lengend Snippet: 4 × 10 4 SHZ-88 cells were implanted into the bone marrow cavity of the hind tibia of rats, which was administered orally with 15 mg/kg/day Syringetin for 21 days. (A) Histopathological status of rat tibia tissue stained with HE (100× and 400×). (B) AS scores and PWT scores. (C) WB detection of ESR1 and PRDM2 expression in tibial bone marrow tissue. N = 9 replicates/group. ESR1, estrogen receptor 1; HE, hematoxylin-eosin; AS, Ambulatory score; PWT, paw withdrawal threshold; WB, Western blotting; PRDM2, positive regulatory domain zinc finger protein2. *p < 0.05 vs . Sham, # p < 0.05 vs . BCP.

Article Snippet: Then, first antibodies were added onto the membrane, including ESR1 (21244-1-AP, 1:2,000, Proteintech), PRDM2 (27718-1-AP, 1:500, Proteintech) and β-actin ( AWA80002 , 1:5,000, Abiowell).

Techniques: Staining, Expressing, Western Blot

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