Perineuronal oligodendrocyte and myelin renewal are epigenetically silenced contributing to cognitive deficits in a murine model of schizophrenia

Perineuronal oligodendrocyte and myelin renewal are epigenetically silenced contributing to cognitive deficits in a murine model of schizophrenia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Perineuronal oligodendrocyte and myelin renewal are epigenetically silenced contributing to cognitive deficits in a murine model of schizophrenia Xianjun Chen, Yuhao Cai, Yao Chen, Mei Li, Mengyao Zhao, Min Jiang, and 17 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8000305/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 9 You are reading this latest preprint version Abstract Cognitive deficits are core symptoms of schizophrenia (SCZ) and major contributors to disability. Beyond gray matter loss, significant oligodendrocytes (OLs) and myelin pathologies have been well-identified in SCZ patients. Cognitive processing highly relies on the myelination in the adult brain. However, whether and how OLs and myelin dynamics contribute to disease pathology remains unexplored. Here, we characterized a preferential loss of perineuronal OLs in both the dizocilpine (MK801)-induced mouse model and postmortem SCZ brain tissue. The extent of myelination was decreased in the medial prefrontal cortex (mPFC) of MK801 mice. Strikingly, spontaneous myelin renewal was suppressed due to a global deficiency in H3 lysine 4 trimethylation (H3K4me3) in OLs. Conditional knockout of Setd1a, the specific H3K4 methyltransferase in oligodendroglia precursor cells (OPCs), impaired myelination. Notably, pharmacologically counteracting H3K4me3 deficiency repaired OLs and myelin deficits and promoted cognitive recovery in MK801 mice. Taken together, these results demonstrate H3K4me3 status as a new potential target enhancing myelin repair to alleviate SCZ-related cognitive impairment. Health sciences/Diseases/Psychiatric disorders/Schizophrenia Biological sciences/Neuroscience Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 INTRODUCTION Schizophrenia (SCZ) is a severe psychiatric disorder that affects more than 23 million individuals worldwide. Over the past century, SCZ has changed little in prevalence and disability 1 . Cognitive deficit is an independent, core and enduring symptom of SCZ, separate from both positive and negative symptoms, yet closely interrelated with them 2 . This deficit contributes to poor clinical outcomes such as functional competence, work skills and an inability to live independently 3 , 4 . Currently, antipsychotics are the primary therapeutic approaches for SCZ in clinical practice 5 . However, no significant improvement can be achieved in the treatment of cognitive symptoms with current pharmacotherapy, even though positive symptoms can be significantly alleviated 6 . Therefore, cognitive deficit is a prevailing issue in the pathogenesis of SCZ, thereby highlighting the paradigm shift from symptom management to functional recovery. Oligodendrocytes (OLs) are myelin-forming cells in the central nervous system. Myelin sheaths wrap axons to promote conduction velocity, provide energy supply, allow neuronal circuits to be finely tuned and synchronized and maintain the proper connectivity among brain regions 7 – 9 . In the adult brain, cognitive processing highly relies on the interplay of neuronal activity-dependent existing myelin modifications and new myelin formation 10 , 11 . Accumulating evidence indicates a notable loss of OLs and myelin in SCZ pathology. Transcriptome and genetic architecture revealed significant molecular burden in oligodendrocyte lineage cells 12 , 13 , and myelin abnormalities have been identified in SCZ patient brains 14 . Therefore, promoting myelin repair could be available to restore cognitive impairments in SCZ. In particular, SCZ psychosis emerges between ages 18–25, which overlaps with the time for vigorous OLs and myelin development 15 . After the first onset, it may persist into adulthood or even old age after multiple relapses. To be mentioned, myelination in the vast majority of brain regions is largely complete by middle age, the new myelin is continuously generated throughout life via the differentiation of adult oligodendroglia precursor cells (OPCs) 16 , 17 . The ongoing deposition of OLs and myelin is required for a range of cognitive tasks, including motor skill learning, fear and spatial memory preservation, which is influenced by factors such as neuronal electrical activity and environmental stimuli 17 – 19 . However, whether myelin dynamics is influenced and contribute to cognitive deficits during the pathogenesis of SCZ is lack of evidence. Here, we report a preferential loss of perineuronal OLs (pn-OLs) in MK801-induced mouse model and individuals with SCZ. Strikingly, except for a loss of myelin sheath, we found adult OPC differentiation and new myelin deposition were significantly impaired, which may be due to H3 lysine 4 trimethylation (H3K4me3) deficiency. We confirmed that ablating H3K4 methyltransferase SETD1A in OPCs arrested myelination. While epigenetic rescuing H3K4me3 deficiency repaired pn-OLs and myelin deficits and promote cognitive recovery in MK801 mice. Together, this study identified an epigenetic barrier for OLs generation and myelin production, providing a potential target for myelin repair to improve cognitive impairment. MATERIALS AND METHODS Human postmortem brain tissue and immunostaining Human schizophrenia and healthy comparable post-mortem tissue were provided by the National Health and Disease Human Brain Tissue Resource Center at Zhejiang University in China. The study was performed in accordance with the ethical standards in the Declaration of Helsinki and was approved by the Human Ethics committee of Zhejiang University School of Medicine (Approval Number: 2020–005). All human tissues were collected following fully informed consent by the donors via a prospective donor scheme. Cases information has been described in Supplementary Table 1. The immunohistochemical staining and image capturing have been described previously 20 . Animals The NG2-CreERT (The Jackson Laboratory, strain no. 008538) mice were crossed with Tau-mGFP (The Jackson Laboratory, strain no. 021162) mice to obtain NG2-CreERT; Tau-mGFP mice. To specifically delete Setd1a in OLs, the NG2-CreERT mice were crossed with Setd1a floxed mice (GemPharmatech Co., Ltd, T013202). Different genotypes of mice were identified using PCR genotyping with the respective primers. The C57BL/6 mice (either sex, 8 weeks old) were obtained from Chongqing Medical Animal Experimentation Center. Animal treatment and study protocols were approved by the Ethical Committee of Chongqing Medical University. Tamoxifen and drug administration Tamoxifen (Sigma-Aldrich, catalog no. T5648) dissolved in ethanol and sunflower oil (1:9, v/v) was administered to induce Cre recombination at a concentration of 30 mg/mL. Adult mice were dosed at 30 mg/kg per day for 4 days. MK801 (MCE, catalog no. HY-15084) was dissolved in saline (0.9% NaCl) at a final concentration of 0.1 mg/mL, and mice were intraperitoneally injected with either the saline vehicle (control group, CTL) or MK801 once daily for 7 consecutive days. After the injection, the mice were allowed to deteriorate for an additional 3 weeks (MK801 + 1 month group, MK801). MK801 mice were treated with TCP (Sigma, catalog no. P8511, 3 mg/kg/day) and ORY-1001 (Selleck, catalog no. S7795, 10 µg/kg/day) for 7 days while receiving an injection of MK801. Briefly, EdU (Sigma, catalog no. 900584) was dissolved in saline at a concentration of 1 mg/mL, and mice were received a constantly intraperitoneal injection for 10 days, with a dose of 5 mg/kg per day. Behavior tests Mice were housed in a controlled environment (25℃) with free access to food and water and maintained on a 12-h/12-h light/dark cycle. After each experiment, all the apparatuses were wiped clean with 70% ethanol to prevent a bias due to olfactory cues. For all behavioral experiments, investigators were kept blinded to the treatment groups and mice were gently handled to avoid stress. All of the tests were recorded and analyzed by EthoVision XT 11.5 software and SMART 3.0 software. Y maze Spontaneous alternative Y-maze test is well-accepted for assessing working memory in mice 21 . Briefly, each mouse was gently put in the center of the apparatus (4 × 36 cm each arm, arranged at an angle of 120° to each other, with 15.5 cm-high walls), facing one fixed arm, and then allowed to explore freely in the maze for 8 min. During the trial, the number of arm entries and alternations was calculated and measured. Morris water maze The Morris water maze test was conducted according to a described protocol 22 . During the acquisition phase, the water maze pool (depth 60 cm, diameter 120 cm) was divided into four equal quadrants, with a hidden escape platform (diameter 6 cm) fixed in one of the quadrants. Each mouse was allowed to locate the hidden platform for 5 consecutive days from different quadrants (one quadrant per trial). In probe test, the platform was removed, and each mouse was allowed to explore for 1 min, the time and distance traveled in the target quadrant was recorded. Immunohistochemistry and EdU staining Immunofluorescence staining was performed as follows: floating tissue sections (20 µm) were obtained from brains perfused with 4% paraformaldehyde (PFA). The sections were first immersed in 0.5% Triton X-100 for 30 min, followed by blocking with 5% bovine serum albumin (BSA) for 1 h at room temperature, and then incubated with primary antibodies overnight at 4℃. On the following day, tissue sections were incubated with secondary antibodies for 1 h at room temperature. Primary antibodies include: ASPA (1:500, oasis, catalog no. OB-PRT005), MBP (1:500, oasis, catalog no. OB-PRT123), SETD1A (1:200, abcam, catalog no. ab243881), PDGFRα (1:300, R&D, catalog no. AF1062), NeuN (1:500, abcam, catalog no. ab104224), H3K4me3 (1:1000, abcam, catalog no. ab8580), GFP (1:1000, abcam, catalog no. ab5450). EdU staining was strictly performed following the manufacturer’s instruction (Beyotime, catalog no. C0081). Western blotting Western blotting Proteins of frozen samples were extracted using RIPA lysis buffer (Beyotime, catalog no. P0013C), which contained protease inhibitors (PMSF, Beyotime, catalog no. ST506). The extracted proteins were then denatured in gel-loading buffer (Beyotime, catalog no. P0286), and 40 µg of protein was separated by 6% and 12% SDS-PAGE gels. The separated proteins were transferred onto PVDF membranes (Millipore, catalog no. IPVH00010), and visualized via enhanced chemiluminescence (ECL) (BIO-OI, catalog no. OI900, China) detection. Band intensity quantification was conducted with Image-Pro Plus software 5.0 (Media Cybernetics). Primary antibodies include: TBP (1:2000, CST, catalog no. 8515), MBP (1:1000), SETD1A (1:5000). RNA extraction and analysis The RNA isolation and reverse transcription were performed as previously described 23 . The total RNA was extracted using TRIzol (Invitrogen, catalog no. 15596026), and reverse transcription was conducted following the manufacturer’s instruction (Vazyme, catalog no. R233-01). The primers for quantitative RT-PCR (RT-qPCR) are as follows: SETD1A: forward 5’-CCCATTAGCAAGAAGGAGAAAGA-3’, reverse 5’-GCTCAGAAAGCACCCGATTA − 3’; WDR5: forward 5’-GGCCACAGTAACTACGTCTTC-3’, reverse 5’-CGTCCCATATCCTCACACTTTC-3’; KMT2D: forward 5’-GTTCGGATAGGAAGGAGCTTATG-3’, reverse 5’-CATCCTCAAGCCCTGGTAAAT-3’; KDM5A: forward 5’-CGTGCCTATCACTCTGGATTTA-3’, reverse 5’-CTGAGCCGTCGATAGTGATTT-3’; KDM5B: forward 5’-CAAGAGCCCACTGAGAAGAAA-3’, reverse 5’-TCCACATAAGAGGCACACATAC-3’; MBP: forward 5’-CTCAGAGGACAGTGATGTGTTT-3’, reverse 5’-CGCCTTGCCAGTTATTCTTTG-3’. Transmission electron microscopy The medial prefrontal cortex (mPFC) samples were perfused with 2.5% glutaraldehyde (dissolved in 4% PFA, Macklin, catalog no. G810415). Subsequent detection and capture of these samples were conducted at the Electron Microscopy Center of Chongqing Medical University. The images of these sections were analyzed using ImageJ-Fiji software (version 1.52p). Single-nucleus transcriptome and sequencing The mPFC samples were rapidly frozen in Liquid nitrogen. Subsequent single-nucleus RNA-seq experiment (snRNA-seq) was conducted at the Key Laboratory of Molecular Biology on Infectious Diseases, Chongqing Medical University. a droplet-based microfluidics platform adapted from the inDrop-seq methodology 24 . Briefly, single-nucleus suspensions were co-encapsulated with barcoded, degradable hydrogel beads in nanoliter-droplets. Within each droplet, bead degradation released primers to capture polyadenylated mRNA. This was immediately followed by reverse transcription and template switching using a Template Switching Oligo (TSO) to generate barcoded, full-length cDNA. The pooled cDNA was then collected for library preparation. The Seurat package (version:5.0.2) was used for cell normalization and regression based on the expression table, according to the UMI counts of each sample and percent of mitochondria rate to obtain the scaled data. Principal component analysis (PCA) was constructed based on the scaled data. The top 2000 high variable genes and top 15 principals were used for uniform manifold approximation and projection (UMAP) construction. Using the graph-based cluster method, we acquired the unsupervised cell cluster result based on the PCA top 15 principals and calculated the marker genes by FindAllMarkers. Significance was defined by the p -value and false discovery rate (FDR). To identify differentially expressed genes among samples, the function FindMarkers with Wilcox rank sum test algorithm was used under the following criteria: Log2FC > 0.25; p -value 0.1. To enable a systematic analysis of cell-cell communication molecules, cellchat was used to identify cell-to-cell interactions in our dataset. Significant mean and cell communication significance ( p -values < 0.05) was calculated based on the interaction and the normalized cell matrix achieved by Seurat normalization. Image acquisition and quantification Fluorescent images were acquired using a confocal laser-scanning microscope (Olympus FV3000 and Nikon NIS-Elements) at appropriate excitation wavelengths. For statistical analysis, the area of the mPFC was measured in each sample for quantification, and was performed using ImageJ-Fiji software (version 1.52p). Statistics Statistical analyses were performed using GraphPad Prism 9 software. All values were presented as mean ± SEM. Between-group comparisons were analyzed using two-tailed unpaired t -tests. Data from the acquisition phase of the Morris water maze test was analyzed using repeated-measures two-way ANOVA with separate analyses for each parameter. Each experiment was performed with at least three independent samples. Statistical significance was defined as a p -value < 0.05. RESULTS Preferential loss of perineuronal oligodendrocytes in MK801-induced mouse model and individuals with schizophrenia Glutamatergic hypoactivity, particularly decreased N-Methyl-D-Aspartate (NMDA) receptor function, is one of the most accepted pathophysiological hypotheses for cognitive decline in SCZ 25 . To study the pathophysiology of SCZ, we employed MK801-induced mice model, which is a widely-used SCZ model (Fig. 1 A). We firstly verified the prolonged effect of MK801 on cognitive function by analyzing the behavioral outcomes. A lower alteration ratio was found in MK801 mice when they spontaneously traveled in the three arms of Y-maze test, although the number of arm entries was comparable (Fig. 1 B), indicating an impairment of spatial memory. Additionally, MK801 mice were trained to locate a hidden platform in a water maze test. Over the 4-day training period, MK801 mice exhibited longer escape latencies, indicating an impaired spatial learning. Moreover, during the probe test, MK801 mice displayed significantly fewer crossings into the target quadrant compared to controls, suggesting an impaired spatial memory (Fig. 1 C). To be mentioned, these results may not be due to abnormal locomotor activity, as MK801 mice exhibited similar levels of activity by traveling a similar distance in the probe test (Fig. 1 C). Moreover, impaired locomotor activity, depressive and anxious phenotypes were excluded in MK801 mice through different behavioral tests. (Supplementary Fig. 1A-D). In addition, social interaction test illustrated that no significant difference was found in social cognition between control and MK801 mice (Supplementary Fig. 1E). These results indicate that MK801 model reproduces certain aspects of cognitive dysfunction associated with schizophrenia. To verify the alterations of oligodendroglia lineage cells, we analyzed the cell density and found that ASPA + mature OLs (MOLs) instead of PDGFRα + OPCs were decreased in mPFC of MK801 mice (Fig. 1 D-E). In this study, the relative positional relationship between pn-OLs and neurons was validated through immunostaining and 3D reconstruction (Fig. 1 F). Electron microscopy analysis further confirmed the attachment of pn-OL could form long curved or concave impressions on the contacting neuron (Fig. 1 G). We confirmed that 29.2% of ASPA + MOLs are pn-OLs in mPFC of adult mice at the age of 3 months (Fig. 1 H). More strikingly, we found a significant decrease of pn-OLs rather than non-pn-OLs in the mPFC of MK801 mice (Fig. 1 I). To further validate that the loss of pn-OLs is a prominent feature in SCZ, we examined the density of Olig2 + oligodendroglia lineage cells in the cortex of SCZ patients and age-matched controls. We identified a preferential loss of Olig2 + pn-OLs in cortex from SCZ patients (Fig. 1 J). Extensive myelin loss and arrested new myelin deposition in MK801 mice In fact, the morphology of pn-OLs shows no substantial difference from that of typical myelinating OLs 26 , 27 . Thus, we questioned whether loss of pn-OLs could result in myelin deficits. We found that MBP labeled myelin fibers significantly decreased in mPFC of MK801 mice as compared to control mice (Fig. 2 A). To access more details of myelin ultrastructure, we analyzed the organization of myelin sheaths using a transmission electron microscope. Consistently, we found the myelin sheaths in the mPFC of MK801 mice were thinner than those in control mice (Fig. 2 B). It is important to note that the densities of NeuN-positive neurons and NF200-positive axons were not altered in the brains after MK801 treatment (Supplementary Fig. 2A-B). We examined axonal degeneration by labeling for SMI-32P, and axonal degeneration was not apparent in the mPFC of MK801 mice (Supplementary Fig. 2B). These results suggest that MK801 treatment led to a loss of pn-OLs and robust myelin deficiency without causing evident neuronal degeneration. To directly assess myelin generation capacity in mPFC, we further employed the NG2-CreERT; Tau-mGFP mouse line to label newly formed myelin sheaths 28 . As the expression level of Tau is high in MOLs but low in OPCs, membrane-bound GFP (mGFP) accumulates only in newly-differentiated OLs and the newly-formed myelin sheaths formed after tamoxifen administration (Fig. 2 C). Notably, abundant mGFP-positive myelin sheaths were observed in the mPFC of adult mice 28 days post-induction, indicating robust new myelin deposition within one month. In contrast, mGFP-positive sheaths were sparse in the mPFC of MK801 mice (Fig. 2 D). In addition, we found the generation of mGFP + new pn-OLs was significantly impaired in mPFC of MK801 mice compared to control mice. Moreover, newly-generated pn-OLs exhibited a lack of myelin production capacity after MK801 treatment (Fig. 2 E). These results confirmed that the generation of pn-OLs and new myelin were significantly arrested in MK801 mice. Adult oligodendroglia precursor cells differentiation was impaired in MK801 mice MK801 treatment is associated with augmented glutamate release in the mPFC, resembling findings in first-episode schizophrenia 29 . In our study, we also observed an increase in synaptic vesicles within presynaptic bouton of excitatory glutamatergic neuron in mPFC of MK801 mice (Supplementary Fig. 3). To delve deeper into the potential molecular alterations affected by MK801, we profiled 40 089 nuclei from 6 mPFC tissues. Unsupervised clustering of the transcriptomic data revealed distinct populations of all major brain cell types (Supplementary Fig. 4A). Those clusters were annotated as excitatory neurons ( Camk2a , Slc17a7 and Satb2 + ), inhibitory neurons ( Pvalb , Vip , Gad1 and Gad2 + ), astrocytes ( Aldh1l1 and Aqp4 + ), OLs ( Plp1 , Mbp , Mobp , Pdgfrα , Cspg4 and Gpr17 + ), endothelial cells ( Rgs5 and Cldn5 + ), and microglia ( Cx3cr1 , C1qb and Tmem119 + ) (Supplementary Fig. 4B). We then compared single-nuclei transcriptional profiles of 6 clusters including oligodendroglia lineage cells from the MK801 and CTL mice (Supplementary Fig. 4C, Supplementary Table 2). Cluster analysis of OLs further revealed three subclusters: OPCs, newly formed OLs (NFOLs), and MOLs, which were based on those established markers including Pdgfrα , Cspg4 , Gpr17 , Enpp6 , Mbp , and Plp1 (Fig. 3 A-C) 30, 31 . Numerous differentially expressed genes (DEGs) were observed across OL subclusters in MK801-treated versus control mice with a pronounced downregulation (Fig. 3 D, Supplementary Table 3). UMAP visualization and cellular proportion analysis demonstrated a significant increase of OPCs and a decrease of MOLs in the MK801-treated group compared to controls (Fig. 3 E), implying an impaired OPCs differentiation. Previous studies have found that functional synapses are formed between OPCs and glutamatergic neurons, enabling OPCs to perceive and decode neuronal activity 32 . We applied a predictive tool to explore the relationship of ligands of excitatory neurons to the receptors expressed in OL subclusters. We found that MK801 treatment disrupted communication mediated by the glutamate signaling between OPCs, NFOLs and excitatory neurons (Supplementary Fig. 4D). Consistently, GO analyses of downregulated genes in OPCs and NFOLs further proved that synapse related cellular components were severely affected in MK801 mice (Fig. 3 F-H). To investigate the influence of aberrant glutamate signaling on OPCs differentiation, we subsequently traced the dividing OPCs using EdU and found the number of ASPA + /EdU + newly differentiated OLs was significantly decreased in the mPFC of MK801 mice (Fig. 3 I). These results suggest MK801-induced aberrant glutamate signaling may impair adult OPCs differentiation. Newly differentiated oligodendrocytes are epigenetically silenced via H3K4me3 insufficiency The acute administration of MK801 causes long-term differentiation barriers in adult OPCs, which implies an endogenous epigenetic abnormality in OLs. In fact, chromatin compaction positively correlates with the maturation stage of OLs 33 . Here we confirmed that euchromatin marked by H3K4me3 (a transcriptionally active chromatin modification), was higher in OPCs and significantly decreased in mature stage (Fig. 4 A-B). We found MOLs within the mPFC of MK801 mice exhibited aberrantly lower or undetectable levels of H3K4me3 compared to that of MOLs in control mice (Fig. 4 C). Notably, the newly-generated MOLs expressed higher levels of H3K4me3 than pre-existing MOLs, implying a higher transcriptional activity. However, a global loss of the active H3K4me3 modification was observed in newly-formed MOLs after MK801 treatment (Fig. 4 D), suggesting an epigenetic suppression in these cells. We further examined the enzymes regulating H3K4me3 status. Strikingly, SETD1A, a key H3K4 methyltransferase, was significantly downregulated in MK801 mice, despite no change in the expression of its functional partners (Fig. 4 E). In contrast, the lysine demethylases KDM5A and KDM5B were significantly upregulated (Fig. 4 F). These results suggest that the H3K4me3 deficiency may set a barrier inhibiting OLs produce myelin in MK801 mice. To confirm the role of H3K4me3 in OLs myelination, we analyzed the single-cell RNA sequencing (scRNA-seq) data of mPFC cells that were freshly isolated from young-adult Setd1a knockout mice 34 . Using a Seurat-based workflow 35 , 3 main clusters of oligodendroglia lineage cells were successfully identified (Supplementary Fig. 5A), including OPCs, NFOLs and MOLs, based on cluster marker genes and known cell type markers (Supplementary Fig. 5B) 30 , 31 . Cell proportion analysis revealed a decreasing trend of NFOLs in mPFC from Setd1a +/− heterozygous mouse (Supplementary Fig. 5C). GO analyses of differentially expressed genes further proved that myelin sheath related cellular components were severely affected by Setd1a deficiency (Supplementary Fig. 5D). To validate the impact of H3K4me3 insufficiency on myelination in vivo , we deleted Setd1a in the OPCs ( Setd1a cKO) (Fig. 5 A). The knockout efficiency of Setd1a was confirmed by the diminished expression of the SETD1A protein in corpus callosum of Setd1a cKO mice (Fig. 5 B). Strikingly, both the transcript and protein levels of MBP, a key component of myelin, significantly reduced in Setd1a cKO mice (Fig. 5 B-C). As expected, SETD1A deficiency led to widespread decrease of H3K4me3 level in OPCs (Fig. 5 D). To further validate that myelin deficiency is a prominent feature, we examined myelin patterns in the mPFC. The areas of MBP + myelinated fibers significantly decreased in mPFC of Setd1a cKO mice compared to wildtype littermates (Fig. 5 E). While the disturbed myelin formation in Setd1a cKO mice was further validated via electron microscopy (Fig. 5 F). These results suggest that H3K4me3 status is essential for OLs myelination. Pharmacological strategy counteracting H3K4me3 deficiency restoring myelin deficits and cognitive dysfunction in MK801 mice. Given the therapeutic implications of these results, we investigated whether counteracting H3K4me3 deficiency would enable OL myelin-producing programs to proceed and therefore improve cognition in MK801 mice. A previous study has proved that lysine specific demethylase 1 (LSD1), a demethylase that counteracts SETD1A, is the most abundant demethylase in the frontal cortex 36 . Building on this finding, we examined the effects of inhibiting LSD1 on myelin deficits with systemic administration of TCP and ORY-1001 for 1 month (Fig. 6 A). As expected, the drug treatment significantly increased the H3K4me3 level in EdU + newly-generated MOLs, as compared with the vehicle group (Fig. 6 B). To be noted, the density of pn-OLs was recovered after those drug treatment (Fig. 6 C). More strikingly, immunostaining showed that the deficiency of MBP + myelinated fibers in the mPFC of MK801 mice was repaired after drug treatment (Fig. 6 D). Electron microscopy further confirmed increased myelin thickness in the mPFC of drug-treated mice compared to vehicle controls (Fig. 6 E). These results suggest that rescuing H3K4me3 deficiency in OLs enhances the generation of pn-OLs and myelin repair in the MK801 brains. We next sought to determine whether in vivo post-developmental LSD1 antagonism by TCP could repair cognitive deficits parallel to enhanced myelination. Mice were subjected to the Y-maze and Morris water maze tasks. MK801 mice treated with TCP and ORY-1001 showed a higher alteration ratio during the Y-maze task, indicating improved working memory compared to vehicle-treated mice (Fig. 6 F). Moreover, over 4 days of training, all groups required progressively less time to reach the platform, reflecting a similar degree of spatial learning. While during the probe test, in which the hidden platform was removed, the drug-treated mice spent more time and traveled a longer distance in the quadrant that previously contained the platform relative to vehicle mice (Fig. 6 G). Thus, systemic treatment with a H3K4me3 targeting drug enhancing myelin repair could improve cognitive performance in MK801 mice. DISCUSSION In this study, we demonstrated a preferential loss of pn-OLs in both SCZ patients and MK801 mice. In addition to widespread myelin loss, we provided the first evidence that endogenous OL and myelin renewal were severely blocked after an acute MK801 treatment, which contributes to cognitive deficits in mice. Moreover, we identified H3K4me3 as a crucial factor governing new myelin deposition. Finally, we proved that pharmacologically rescuing H3K4me3 deficiency significantly enhanced myelin repair, and thereafter improved the cognitive performance of MK801 mice. Cognitive decline may begin in adolescents diagnosed with SCZ, even precede illness onset, and accelerate in adulthood 37 , 38 . Supporting this notion, we employed a mouse model by injecting MK801 in young-adult mice. In fact, the MK801 model was proved to have good performance, construct and predictive validity for studying the pathophysiology of cognitive dysfunction in SCZ 39 . Previous study found that MK801-induced disruption of NMDA receptor function during adolescence can lead to persistent dysfunction of the prefrontal cortex in adulthood 40 . Similarly, we found that an acute MK801 treatment in young-adult mice led to prolonged cognitive deficits, especially the working memory and spatial memory (Fig. 1 ). However, in our study, social cognition as well as emotional alterations was intact in MK801 mice (Supplementary Fig. 1), which highlights the heterogeneity of cognitive impairments in patients. In fact, cognitive heterogeneity not only compromises the prediction of clinical outcomes but also impedes the development of precision treatment strategies. Accumulating evidence indicates that both OLs and myelin are significantly altered during the pathogenesis of schizophrenia. In this study, we clearly identified a preferential loss of pn-OLs in the cortex of SCZ patients as well as the MK801 mouse model (Fig. 1 ). The term of pn-OLs was coined by the pioneering neurohistologist Pío del Río Hortega decades ago 26 , while the function of these cells in the cortex remains unknown. We confirmed that pn-OLs were able to produce myelin, and deficits of adult OPC differentiation and new myelin formation led to myelin deficiency under MK801 treatment (Fig. 2 ). It is known that neuronal activity can affect OPCs differentiation, maturation and myelin formation through the glutamate signaling 41 , 42 . In this study, we found that MK801 treatment disrupted the communication between OPCs/NFOLs and excitatory neurons, which may impair OPCs differentiation (Supplementary Fig. 4). Because our understanding of the functional interplay between OLs and excitatory neurons is still at its infancy, the precise mechanism governing pn-OLs function still remains undefined. Besides myelination, accumulating evidence suggests that pn-OLs can provide metabolic and neurotrophic support to nearby neurons 43 . Interestingly, studies have shown that the localization of pn-OLs exhibits neuronal selectivity, preferentially associating with glutamatergic neurons compared to GABAergic interneurons 26 . What’s more, the pn-OLs express ionotropic and metabotropic glutamate receptors, glutamate transporters and glutamine synthetase (GS) for converting glutamate to glutamine 44 – 47 . These findings further support that pn-OLs appear to be sensitive to glutamate metabolism, which may in turn facilitate the metabolism of neurons and modulate their activity 46 . Impaired adult OPCs differentiation has long been considered the primary cause of remyelination failure 48 . In other cases, even though MOLs are present, they fail to generate new myelin sheaths, which may due to epigenetic barrier 49 . From the insight of development, epigenetic regulation is crucial for OPCs differentiation 50 . Here, after MK801 insult, we found a global loss of the active H3K4me3 modification in the newly-generated MOLs (Fig. 4 ). Impairment of the epigenetic barrier mediated by H3K4me3 compromised the ability of MOLs to produce myelin. This process could be primarily mediated through a series of enzymatic regulation, such as SETD1A, KDM5A and KDM5B (Fig. 4 ). To be noted, Setd1a has recently been identified as a risk gene for SCZ, and Setd1a mutations confer a large increase in disease risk 51 . Recently, multiple studies have elucidated how Setd1a affects neuronal development and synaptic function, and thereafter contribute to the onset of SCZ 34 , 36 , 52 . However, whether Setd1a affects OLs function in SCZ remains unclear. In this study, we provide the first evidence that SETD1A, a key regulator of H3K4me3, is tightly related to OL myelination (Fig. 5 ), shedding light on H3K4me3 status in white matter etiology of SCZ. Numerous studies have explored strategies to enhance myelin formation in disease treatment. Chen, Jing-Fei et al. found that promoting myelinogenesis significantly improved performance in memory-related tasks and alleviated cognitive impairment in a mouse model of Alzheimer's disease 53 . These findings suggest that myelin repair represents a potentially effective strategy for ameliorating cognitive deficits in SCZ. Recently, a number of histone demethylase inhibitors with good blood-brain barrier permeability have been identified. We speculate that such compounds are likely to induce substantial changes in methylation states, thereby counteracting the effects of H3K4me3 deficiency. In this study, we found treatment with two LSD1 inhibitors, TCP and ORY-1001, rescued pn-OLs and myelin deficits and ameliorated cognitive deficits in MK801-treated mice (Fig. 6 ), This is an exciting observation, as it presents a promising target for promoting endogenous remyelination, and thereafter overcome myelin related cognitive deficits in major psychiatric disorders represented by SCZ. In conclusion, our findings identified the pathological potential of pn-OLs, and provided new evidence of myelin generation deficits in SCZ, possibly due to an H3K4me3-mediated epigenetic barrier. Targeting the H3K4me3 status in OLs could be a novel therapeutic strategy for cognitive deficits in SCZ. Declarations DATA AVAILABILITY The data are available in the main text and supplementary materials. The single-nucleus transcriptome statistics are available from the corresponding author, XC, upon reasonable request. ACKNOWLEDGEMENTS This work was supported by the National Natural Science Foundation of China (32130019, 32000684), the National Key Research and Development Program of China (2021YFA1100203) and Foundation of Chongqing Educational Committee (KJQN202500421). AUTHOR CONTRIBUTIONS Conceptualization: XC, BW, and LZ; Methodology: XC and YC; Investigation: XC, YC, YC, ML, MZ, MJ, SL, BT, CZ, XY, AG, QJ, WX, JL, QL, YM, JX, JN and LX; Visualization: XC, YC, ML and YC; Funding acquisition: XC and BW; Project administration: XC and BW; Supervision: XC and BW; Writing–original draft: XC, YC, ML, YC, QL and MJ; Writing–review & editing: XC, YC, ML, YC, QL and MJ. COMPETING INTERESTS The authors declare no competing interests. References Hegarty JD, Baldessarini RJ, Tohen M, Waternaux C, Oepen G. One hundred years of schizophrenia: a meta-analysis of the outcome literature. The American journal of psychiatry 1994; 151(10): 1409–1416. Rodriguez-Jimenez R, Bagney A, Mezquita L, Martinez-Gras I, Sanchez-Morla EM, Mesa N et al. Cognition and the five-factor model of the positive and negative syndrome scale in schizophrenia. Schizophrenia research 2013; 143(1): 77–83. Bowie CR, Leung WW, Reichenberg A, McClure MM, Patterson TL, Heaton RK et al. Predicting schizophrenia patients' real-world behavior with specific neuropsychological and functional capacity measures. Biological psychiatry 2008; 63(5): 505–511. Kane JM, Lencz T. Cognitive deficits in schizophrenia: short-term and long-term. World psychiatry: official journal of the World Psychiatric Association (WPA) 2008; 7(1): 29–30. Efthimiou O, Taipale H, Radua J, Schneider-Thoma J, Pinzón-Espinosa J, Ortuño M et al. Efficacy and effectiveness of antipsychotics in schizophrenia: network meta-analyses combining evidence from randomised controlled trials and real-world data. The lancet Psychiatry 2024; 11(2): 102–111. Monteleone P, Cascino G, Monteleone AM, Rocca P, Rossi A, Bertolino A et al. Prevalence of antipsychotic-induced extrapyramidal symptoms and their association with neurocognition and social cognition in outpatients with schizophrenia in the "real-life". Progress in neuro-psychopharmacology & biological psychiatry 2021; 109: 110250. Fünfschilling U, Supplie LM, Mahad D, Boretius S, Saab AS, Edgar J et al. Glycolytic oligodendrocytes maintain myelin and long-term axonal integrity. Nature 2012; 485(7399): 517–521. Lee Y, Morrison BM, Li Y, Lengacher S, Farah MH, Hoffman PN et al. Oligodendroglia metabolically support axons and contribute to neurodegeneration. Nature 2012; 487(7408): 443–448. Hunt BA, Tewarie PK, Mougin OE, Geades N, Jones DK, Singh KD et al. Relationships between cortical myeloarchitecture and electrophysiological networks. Proceedings of the National Academy of Sciences of the United States of America 2016; 113(47): 13510–13515. Munyeshyaka M, Fields RD. Oligodendroglia are emerging players in several forms of learning and memory. Communications biology 2022; 5(1): 1148. Bacmeister CM, Huang R, Osso LA, Thornton MA, Conant L, Chavez AR et al. Motor learning drives dynamic patterns of intermittent myelination on learning-activated axons. Nature neuroscience 2022; 25(10): 1300–1313. Martins-de-Souza D. Proteome and transcriptome analysis suggests oligodendrocyte dysfunction in schizophrenia. Journal of psychiatric research 2010; 44(3): 149–156. Biological insights from 108 schizophrenia-associated genetic loci. Nature 2014; 511(7510): 421–427. Flynn SW, Lang DJ, Mackay AL, Goghari V, Vavasour IM, Whittall KP et al. Abnormalities of myelination in schizophrenia detected in vivo with MRI, and post-mortem with analysis of oligodendrocyte proteins. Molecular psychiatry 2003; 8(9): 811–820. Insel TR. Rethinking schizophrenia. Nature 2010; 468(7321): 187–193. Hill RA, Li AM, Grutzendler J. Lifelong cortical myelin plasticity and age-related degeneration in the live mammalian brain. Nature neuroscience 2018; 21(5): 683–695. Hughes EG, Orthmann-Murphy JL, Langseth AJ, Bergles DE. Myelin remodeling through experience-dependent oligodendrogenesis in the adult somatosensory cortex. Nature neuroscience 2018; 21(5): 696–706. Pan S, Mayoral SR, Choi HS, Chan JR, Kheirbek MA. Preservation of a remote fear memory requires new myelin formation. Nature neuroscience 2020; 23(4): 487–499. Bacmeister CM, Barr HJ, McClain CR, Thornton MA, Nettles D, Welle CG et al. Motor learning promotes remyelination via new and surviving oligodendrocytes. Nature neuroscience 2020; 23(7): 819–831. Yu G, Su Y, Guo C, Yi C, Yu B, Chen H et al. Pathological oligodendrocyte precursor cells revealed in human schizophrenic brains and trigger schizophrenia-like behaviors and synaptic defects in genetic animal model. Molecular psychiatry 2022; 27(12): 5154–5166. Choi YK, Maki T, Mandeville ET, Koh SH, Hayakawa K, Arai K et al. Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury. Nature medicine 2016; 22(11): 1335–1341. Vorhees CV, Williams MT. Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nature protocols 2006; 1(2): 848–858. Chen X, Ku L, Mei R, Liu G, Xu C, Wen Z et al. Novel schizophrenia risk factor pathways regulate FEZ1 to advance oligodendroglia development. Translational psychiatry 2017; 7(12): 1293. Klein AM, Mazutis L, Akartuna I, Tallapragada N, Veres A, Li V et al. Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. Cell 2015; 161(5): 1187–1201. Buck SA, Quincy Erickson-Oberg M, Logan RW, Freyberg Z. Relevance of interactions between dopamine and glutamate neurotransmission in schizophrenia. Molecular psychiatry 2022; 27(9): 3583–3591. Bernstein HG, Keilhoff G, Dobrowolny H, Guest PC, Steiner J. Perineuronal oligodendrocytes in health and disease: the journey so far. Reviews in the neurosciences 2019; 31(1): 89–99. Takasaki C, Yamasaki M, Uchigashima M, Konno K, Yanagawa Y, Watanabe M. Cytochemical and cytological properties of perineuronal oligodendrocytes in the mouse cortex. The European journal of neuroscience 2010; 32(8): 1326–1336. Wang F, Ren SY, Chen JF, Liu K, Li RX, Li ZF et al. Myelin degeneration and diminished myelin renewal contribute to age-related deficits in memory. Nature neuroscience 2020; 23(4): 481–486. Zou Y, Zhang H, Chen X, Ji W, Mao L, Lei H. Age-dependent effects of (+)-MK801 treatment on glutamate release and metabolism in the rat medial prefrontal cortex. Neurochemistry international 2019; 129: 104503. Hou J, Zhou Y, Cai Z, Terekhova M, Swain A, Andhey PS et al. Transcriptomic atlas and interaction networks of brain cells in mouse CNS demyelination and remyelination. Cell reports 2023; 42(4): 112293. Dennis DJ, Wang BS, Karamboulas K, Kaplan DR, Miller FD. Single-cell approaches define two groups of mammalian oligodendrocyte precursor cells and their evolution over developmental time. Stem cell reports 2024; 19(5): 654–672. Bergles DE, Roberts JD, Somogyi P, Jahr CE. Glutamatergic synapses on oligodendrocyte precursor cells in the hippocampus. Nature 2000; 405(6783): 187–191. Liu J, Magri L, Zhang F, Marsh NO, Albrecht S, Huynh JL et al. Chromatin landscape defined by repressive histone methylation during oligodendrocyte differentiation. The Journal of neuroscience: the official journal of the Society for Neuroscience 2015; 35(1): 352–365. Chen R, Liu Y, Djekidel MN, Chen W, Bhattacherjee A, Chen Z et al. Cell type-specific mechanism of Setd1a heterozygosity in schizophrenia pathogenesis. Sci Adv 2022; 8(9): eabm1077. Butler A, Hoffman P, Smibert P, Papalexi E, Satija R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nature biotechnology 2018; 36(5): 411–420. Mukai J, Cannavò E, Crabtree GW, Sun Z, Diamantopoulou A, Thakur P et al. Recapitulation and Reversal of Schizophrenia-Related Phenotypes in Setd1a-Deficient Mice. Neuron 2019; 104(3): 471–487.e412. Lee M, Cernvall M, Borg J, Plavén-Sigray P, Larsson C, Erhardt S et al. Cognitive Function and Variability in Antipsychotic Drug-Naive Patients With First-Episode Psychosis: A Systematic Review and Meta-Analysis. JAMA psychiatry 2024; 81(5): 468–476. Jonas K, Lian W, Callahan J, Ruggero CJ, Clouston S, Reichenberg A et al. The Course of General Cognitive Ability in Individuals With Psychotic Disorders. JAMA psychiatry 2022; 79(7): 659–666. Brown JW, Rueter LE, Zhang M. Predictive validity of a MK-801-induced cognitive impairment model in mice: implications on the potential limitations and challenges of modeling cognitive impairment associated with schizophrenia preclinically. Progress in neuro-psychopharmacology & biological psychiatry 2014; 49: 53–62. Flores-Barrera E, Thomases DR, Tseng KY. MK-801 Exposure during Adolescence Elicits Enduring Disruption of Prefrontal E-I Balance and Its Control of Fear Extinction Behavior. The Journal of neuroscience: the official journal of the Society for Neuroscience 2020; 40(25): 4881–4887. Li C, Xiao L, Liu X, Yang W, Shen W, Hu C et al. A functional role of NMDA receptor in regulating the differentiation of oligodendrocyte precursor cells and remyelination. Glia 2013; 61(5): 732–749. Gautier HO, Evans KA, Volbracht K, James R, Sitnikov S, Lundgaard I et al. Neuronal activity regulates remyelination via glutamate signalling to oligodendrocyte progenitors. Nature communications 2015; 6: 8518. Du Y, Dreyfus CF. Oligodendrocytes as providers of growth factors. Journal of neuroscience research 2002; 68(6): 647–654. Káradóttir R, Cavelier P, Bergersen LH, Attwell D. NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. Nature 2005; 438(7071): 1162–1166. DeSilva TM, Kabakov AY, Goldhoff PE, Volpe JJ, Rosenberg PA. Regulation of glutamate transport in developing rat oligodendrocytes. The Journal of neuroscience: the official journal of the Society for Neuroscience 2009; 29(24): 7898–7908. Yamazaki Y, Hozumi Y, Kaneko K, Fujii S. Modulatory Effects of Perineuronal Oligodendrocytes on Neuronal Activity in the Rat Hippocampus. Neurochemical research 2018; 43(1): 27–40. D'Amelio F, Eng LF, Gibbs MA. Glutamine synthetase immunoreactivity is present in oligodendroglia of various regions of the central nervous system. Glia 1990; 3(5): 335–341. Kuhlmann T, Miron V, Cui Q, Wegner C, Antel J, Brück W. Differentiation block of oligodendroglial progenitor cells as a cause for remyelination failure in chronic multiple sclerosis. Brain: a journal of neurology 2008; 131(Pt 7): 1749–1758. Liu X, Xin DE, Zhong X, Zhao C, Li Z, Zhang L et al. Small-molecule-induced epigenetic rejuvenation promotes SREBP condensation and overcomes barriers to CNS myelin regeneration. Cell 2024; 187(10): 2465–2484.e2422. Emery B, Lu QR. Transcriptional and Epigenetic Regulation of Oligodendrocyte Development and Myelination in the Central Nervous System. Cold Spring Harbor perspectives in biology 2015; 7(9): a020461. Singh T, Kurki MI, Curtis D, Purcell SM, Crooks L, McRae J et al. Rare loss-of-function variants in SETD1A are associated with schizophrenia and developmental disorders. Nature neuroscience 2016; 19(4): 571–577. Su X, Zhang H, Hong Y, Yang Q, Wang L, Le T et al. Mutations of schizophrenia risk gene SETD1A dysregulate synaptic function in human neurons. Molecular psychiatry 2025. Chen JF, Liu K, Hu B, Li RR, Xin W, Chen H et al. Enhancing myelin renewal reverses cognitive dysfunction in a murine model of Alzheimer's disease. Neuron 2021; 109(14): 2292–2307.e2295. Additional Declarations The authors have declared there is NO conflict of interest to disclose Supplementary Files SupplementaryTable1.Informationofhumanpostmortembraintissues.docx Supplementary Table 1 SupplementaryTable2.DifferentialexpressiongenesinmajorcelltypesinmPFCofmice.xlsx Supplementary Table 2 SupplementaryTable3.DifferentialexpressiongenesinoligodendroglialineagecellsinmPFCofmice.xlsx Supplementary Table 3 10.31Caietal.SupplementaryInformation.docx Supplementary Information (Supplementary methods and Supplementary Figure legends) Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: revise 21 Apr, 2026 Review # 2 received at journal 10 Apr, 2026 Reviewer # 2 agreed at journal 16 Mar, 2026 Review # 1 received at journal 13 Mar, 2026 Reviewer # 1 agreed at journal 13 Mar, 2026 Reviewers invited by journal 02 Feb, 2026 Editor assigned by journal 04 Nov, 2025 Submission checks completed at journal 04 Nov, 2025 First submitted to journal 31 Oct, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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spontaneous alteration test. Bar plot showing the total number of entries into the 3 arms and the alteration rate of entry into arms different from the previous arm for CTL (n=8) and MK801 (n=9) mice. \u003cstrong\u003eC.\u003c/strong\u003e Pattern diagram of the Morris water maze test. The latency to platform in the acquisition phase (two-way repeated ANOVA was used), measurement of time and distance spent in the target quadrant, and the total distance in quadrant after platform removal (n=14 mice for each group). \u003cstrong\u003eD.\u003c/strong\u003e and \u003cstrong\u003eE.\u003c/strong\u003e Representative images showing immunofluorescence staining and quantification of PDGFRα\u003csup\u003e+\u003c/sup\u003e (red) and ASPA\u003csup\u003e+\u003c/sup\u003e (green) cells in the mPFC of mice. Scale bars, 100 μm (left panels) and 20 μm (right panels). \u003cstrong\u003eF.\u003c/strong\u003e Representative images showing perineuronal MOL (ASPA\u003csup\u003e+\u003c/sup\u003e, red) and Neuron (NeuN\u003csup\u003e+\u003c/sup\u003e, green) in the mPFC of mice and 3D reconstruction. Scale bars, 10 μm. \u003cstrong\u003eG.\u003c/strong\u003e Electron micrographs showing pn-OL tightly contacting the soma of a neuron. Scale bars, 2 μm (left panel) and 500 nm (right panel). \u003cstrong\u003eH.\u003c/strong\u003e Pie chart showing the distribution of two groups (n=5 for CTL group). \u003cstrong\u003eI.\u003c/strong\u003e Representative images showing immunofluorescence staining via 3D reconstruction of ASPA\u003csup\u003e+\u003c/sup\u003e (red) and NeuN\u003csup\u003e+\u003c/sup\u003e (green) cells in the mPFC of mice. Scale bars, 50μm. Bar plot showing quantification of the density of pn-OLs and non-pn-OLs in mPFC from CTL and MK801 groups (n=5 mice for each group). \u003cstrong\u003eJ.\u003c/strong\u003e Representative images showing immunohistochemistry of Olig2\u003csup\u003e+\u003c/sup\u003e pn-OLs (black arrow head) and non-pn-OLs (black triangles) in the cortex from schizophrenia patients and healthy controls (n=3 for each group). Scale bar, 10μm. And pattern diagram of pn-OL. Bar plot showing proportion of pn-OLs and non-pn-OLs (Olig2\u003csup\u003e+\u003c/sup\u003e, brown), used paired \u003cem\u003et\u003c/em\u003e-test. Unpaired \u003cem\u003et\u003c/em\u003e-test was used in other plots, and data are expressed as the mean ± SEM, *\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05, or ***\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.001.\u003c/p\u003e","description":"","filename":"Figure11.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/a0d07065a1602d02d0e9c93f.png"},{"id":101863311,"identity":"164baf97-4b56-440e-8540-64a267c5ad7b","added_by":"auto","created_at":"2026-02-04 12:06:01","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":5311983,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMyelin loss and impaired new-myelin formation in mPFC of MK801 mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA. \u003c/strong\u003eRepresentative images showing the expression of MBP (red) in the mPFC of mice (n=6 for each group). Scale bars, 100 μm (left panels) and 20μm (right panels). \u003cstrong\u003eB.\u003c/strong\u003e Electron micrographs showing myelinated axons in mPFC and magnified images in CTL and MK801 groups. Scale bars, 1 μm (left panels) and 0.5 μm (right panels). And quantification of scatter plot of \u003cem\u003eg\u003c/em\u003e-ratio (n=3 for each group). \u003cstrong\u003eC.\u003c/strong\u003e Schematic showing the time course of tamoxifen induction and histology for NG2-CreERT; Tau-mGFP mice and genetic construction. The schematic illustration showing the mGFP expression pattern in the NG2-CreERT; Tau-mGFP mice. \u003cstrong\u003eD. \u003c/strong\u003eRepresentative images and quantification of newly-formed myelin sheath (mGFP\u003csup\u003e+\u003c/sup\u003e, green) in mPFC of mice (n=5 for CTL group, n=6 for MK801 group). Scale bar, 100 μm. \u003cstrong\u003eE.\u003c/strong\u003e Representative images showing the mature pn-OLs (ASPA\u003csup\u003e+\u003c/sup\u003e, red, yellow arrow head), neuron (NeuN\u003csup\u003e+\u003c/sup\u003e, white), nucleus (DAPI, blue) and myelin sheath (mGFP\u003csup\u003e+\u003c/sup\u003e, green) segregated from individual pn-OL for CTL and MK801 groups. Scale bar, 20 μm. Statistics showing density of newly formed pn-OL (n=6 for CTL group, n=5 for MK801 group) and myelin sheath separated from each pn-OL (n=6 mice for each group). Unpaired \u003cem\u003et\u003c/em\u003e-test was used, and data are expressed as the mean ± SEM, *\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/3f66780f956eef94eb1de4a9.png"},{"id":101863308,"identity":"1696fa17-c540-4f05-9a28-7a0f69a5e143","added_by":"auto","created_at":"2026-02-04 12:06:01","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1432487,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSingle-nucleus sequencing and tracing adult OPCs differentiation in mPFC of mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA.\u003c/strong\u003e Unbiased UMAP of measured 3 different cell clusters (OPC, oligodendrocyte precursor cell; NFOL, newly formed oligodendrocyte; MOL, mature oligodendrocyte) between two groups (n=3 per group), each dot representing a single cell, color-coded by cell type. \u003cstrong\u003eB.\u003c/strong\u003e Expression of key marker genes of 3 cell clusters. Color and size bar reflects the percentage of cells expressing each gene within the cluster, while color represents average log-normalised gene expression. \u003cstrong\u003eC.\u003c/strong\u003e Projection of key marker genes (\u003cem\u003ePdgfrα\u003c/em\u003e, \u003cem\u003eCspg4\u003c/em\u003e, \u003cem\u003eGpr17\u003c/em\u003e, \u003cem\u003eEnpp6\u003c/em\u003e, \u003cem\u003eMbp\u003c/em\u003e and \u003cem\u003ePlp1\u003c/em\u003e) onto the graph plot of 3 clusters. Color bar on the right side of each plot represents color scale level. \u003cstrong\u003eD.\u003c/strong\u003e Bar graph showing the number of DEGs of MK801 mice. \u003cstrong\u003eE.\u003c/strong\u003e Bar plot showing the relative distribution of each cluster of two groups. \u003cstrong\u003eF., G. and H.\u003c/strong\u003e Bar graph showing the GO terms enriched in the DEGs of OPC, NFOL and MOL separately in cellular components. \u003cstrong\u003eI. \u003c/strong\u003eSchematic showing EdU injection strategy and timeline. And representative images and quantification of ASPA\u003csup\u003e+\u003c/sup\u003e and EdU\u003csup\u003e+\u003c/sup\u003e newly born MOLs in CTL and MK801 mice (n=6 for CTL group, n=5 for MK801 group). Scale bar, 20 μm.\u003cstrong\u003e \u003c/strong\u003eUnpaired \u003cem\u003et\u003c/em\u003e-test was used, and data are expressed as the mean ± SEM, *\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/0ab48c9c029b597c34d27a24.png"},{"id":101881085,"identity":"6055ddb9-6378-410a-ad51-e814c250a63f","added_by":"auto","created_at":"2026-02-04 15:09:25","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":3867147,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnalysis of H3K4me3 status in OLs from mPFC of mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA.\u003c/strong\u003e Representative ultrahigh-resolution images showing non-overlaps of H3K4me3 (red) and nuclear heterochromatin (bright foci in DAPI, blue) in MOL (ASPA\u003csup\u003e+\u003c/sup\u003e, white). Scale bar, 5 μm. \u003cstrong\u003eB.\u003c/strong\u003e Representative images of H3K4me3 (red) and quantification of integrated density in MOLs (ASPA\u003csup\u003e+\u003c/sup\u003e, white, yellow triangles) and OPCs (PDGFRα\u003csup\u003e+\u003c/sup\u003e, green, white arrow head) in mPFC on P28 CTL group (n=4). Scale bar, 20 μm. \u003cstrong\u003eC.\u003c/strong\u003e Representative images of H3K4me3 (red) in MOLs (ASPA\u003csup\u003e+\u003c/sup\u003e, green) in mPFC. Scale bar, 20 μm. Violin plot and bar plot showing quantification of integrated density cells (n=171 cells for CTL group, n=58 cells for MK801 group) and mice (n=6 mice for each group). \u003cstrong\u003eD. \u003c/strong\u003eRepresentative images of H3K4me3 (red) and quantification of integrated density in mGFP\u003csup\u003e+\u003c/sup\u003e newly formed MOLs and mGFP\u003csup\u003e-\u003c/sup\u003e pre-existing MOLs (n=5 mice for each group). Scale bars, 10 μm (left panels), and 5 μm (right panels). \u003cstrong\u003eE.\u003c/strong\u003e Quantification of mRNA transcripts of H3K4me3 (SETD1A, WDR5, KMT2D) enzyme of mice (n=6 for each group). \u003cstrong\u003eF.\u003c/strong\u003e Quantification of mRNA transcripts of H3K4me3 (KDM5A, KDM5B) enzyme of mice (n=6 for each group). Unpaired \u003cem\u003et\u003c/em\u003e-test was used, and data are expressed as the mean ± SEM, *\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05, **\u003cem\u003e p\u003c/em\u003e \u0026lt; 0.01, ***\u003cem\u003e p\u003c/em\u003e \u0026lt; 0.001, or ****\u003cem\u003e p\u003c/em\u003e \u0026lt; 0.0001.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/2ccf8083f75d500d23de8ca1.png"},{"id":101881901,"identity":"07558777-5718-4c86-bfb6-1ec28950c425","added_by":"auto","created_at":"2026-02-04 15:17:29","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":2882148,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eConditional knockout of Setd1a in OPC lead to myelin deficits.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e. Schematic showing the time course of NG2-CreERT; Setd1a-flox mice. \u003cstrong\u003eB.\u003c/strong\u003e Western blotting showing immunoblots and quantification of SETD1A and MBP in \u003cem\u003eSetd1a\u003c/em\u003e cKO mice and littermates in corpus callosum (n=4 for each group). \u003cstrong\u003eC. \u003c/strong\u003eQuantification of mRNA transcripts of MBP of mice in PFC (n=4 for each group). \u003cstrong\u003eD.\u003c/strong\u003e Representative images showing immunofluorescence staining of H3K4me3 (red) and quantification of integrated density in OPC (PDGFRα\u003csup\u003e+\u003c/sup\u003e, green) in mPFC of WT and \u003cem\u003eSetd1a\u003c/em\u003e cKO mice (n=4 for each group). Scale bars, 20 μm (left panel), 5μm (right panel). \u003cstrong\u003eE.\u003c/strong\u003e Representative images showing immunofluorescence staining and quantification of MBP (red) in mPFC of WT and\u003cem\u003e Setd1a \u003c/em\u003ecKO mice (n=4 for each group). Scale bars, 100 μm (left panel), 50μm (right panel). \u003cstrong\u003eF. \u003c/strong\u003eElectron micrographs showing myelinated axons in optic nerves and magnified images in WT and \u003cem\u003eSetd1a\u003c/em\u003e cKO groups. Scale bars, 1 μm (left panel) and 0.5 μm (right panel). And quantification of scatter plot of \u003cem\u003eg\u003c/em\u003e-ratio (n=3 for each group). Unpaired \u003cem\u003et\u003c/em\u003e-test was used, and data are expressed as the mean ± SEM, *\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05, or **\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.01.\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/5dd21fbe81dcb4f3a435187e.png"},{"id":101881872,"identity":"c2af16af-4b07-4cae-a337-024881c5e010","added_by":"auto","created_at":"2026-02-04 15:17:26","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":4558930,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEpigenetic-targeting compounds enhance myelination and rescues cognitive decline in MK801 mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eA. \u003c/strong\u003eSchematic showing examining the effect of inhibition of lysine specific demethylase 1 (LSD1) on cognitive deficits following systemic ORY-1001 and tranylcypromine (TCP) administration. (CTL group, MK801+VEH group, MK801+TCP group, MK801+ORY group). \u003cstrong\u003eB.\u003c/strong\u003e Representative images of H3K4me3 (red) and quantification of integrated density in EdU\u003csup\u003e+\u003c/sup\u003e (white, white arrow head) newly formed MOLs (ASPA\u003csup\u003e+\u003c/sup\u003e, green) in mPFC (n=5 mice for each group). Scale bars, 20 μm (upward panel) and 5 μm (downward panel). \u003cstrong\u003eC. \u003c/strong\u003eRepresentative images showing immunofluorescence staining via 3D reconstruction of ASPA\u003csup\u003e+\u003c/sup\u003e (green) and NeuN\u003csup\u003e+\u003c/sup\u003e (red) cells in the mPFC of mice. Scale bars, 100 μm. Bar plot showing quantification of the density of pn-OLs in mPFC from groups (n=5 mice for CTL and MK801+VEH groups, n=6 mice for MK801+TCP and MK801+ORY groups). \u003cstrong\u003eD.\u003c/strong\u003e Representative images showing immunofluorescence staining and quantification of MBP in the mPFC of mice (n=5 for each group). Scale bar, 100 μm. \u003cstrong\u003eE. \u003c/strong\u003eElectron micrographs showing myelinated axons in mPFC and magnified images in CTL, MK801+VEH, MK801+TCP and MK801+ORY groups. Scale bar, 1 μm. And quantification of scatter plot and bar plot of \u003cem\u003eg\u003c/em\u003e-ratio (n=3 for each group). \u003cstrong\u003eF. \u003c/strong\u003ePattern diagram of Y-maze spontaneous alteration test. The alteration rate of entry into arms different from the previous arm for CTL (n=9), MK801+VEH (n=10), MK801+TCP and MK801+ORY (n=11) group. \u003cstrong\u003eG.\u003c/strong\u003e Pattern diagram of the Morris water maze test. The latency to platform in the acquisition phase (two-way repeated ANOVA was used), and measurement of time and distance spent in the target quadrant, and the total distance in quadrant after platform removal (n=9 for CTL group, n=10 for MK801+VEH and MK801+TCP group, n=11 for MK801+ORY group). Unpaired \u003cem\u003et\u003c/em\u003e-test was used, and data are expressed as the mean ± SEM, *\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05, or **\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.01.\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/46ee1bba5787e3955c3ec7c8.png"},{"id":101882709,"identity":"e7d78fa8-753a-4b68-bd72-4a351c437db0","added_by":"auto","created_at":"2026-02-04 15:24:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":23585657,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/5dcded32-d060-4bdc-b6d5-44d6a5c05e44.pdf"},{"id":101863302,"identity":"dab7ecb6-b2c5-4ba6-80a8-46e409d173ce","added_by":"auto","created_at":"2026-02-04 12:06:01","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":15697,"visible":true,"origin":"","legend":"Supplementary Table 1","description":"","filename":"SupplementaryTable1.Informationofhumanpostmortembraintissues.docx","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/fece92b21f00fd2ad5552fc3.docx"},{"id":101863312,"identity":"5f7c7309-e4b6-44cd-99d6-2296747aaf96","added_by":"auto","created_at":"2026-02-04 12:06:02","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":61731,"visible":true,"origin":"","legend":"Supplementary Table 2","description":"","filename":"SupplementaryTable2.DifferentialexpressiongenesinmajorcelltypesinmPFCofmice.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/69d0cd129d5ae9cc511d5f5f.xlsx"},{"id":101881990,"identity":"76d1b5cd-e2a2-40c8-8885-0ae7333f5620","added_by":"auto","created_at":"2026-02-04 15:18:02","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":53157,"visible":true,"origin":"","legend":"Supplementary Table 3","description":"","filename":"SupplementaryTable3.DifferentialexpressiongenesinoligodendroglialineagecellsinmPFCofmice.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/c8e656ec44dbc4e20ffef81d.xlsx"},{"id":101863305,"identity":"bc2623df-9b57-4cbb-a252-6036bfefff0b","added_by":"auto","created_at":"2026-02-04 12:06:01","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":30502,"visible":true,"origin":"","legend":"Supplementary Information (Supplementary methods and Supplementary Figure legends)","description":"","filename":"10.31Caietal.SupplementaryInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-8000305/v1/beaa266b07f4a4ea6fb1d361.docx"}],"financialInterests":"The authors have declared there is \u003cb\u003eNO\u003c/b\u003e conflict of interest to disclose","formattedTitle":"Perineuronal oligodendrocyte and myelin renewal are epigenetically silenced contributing to cognitive deficits in a murine model of schizophrenia","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eSchizophrenia (SCZ) is a severe psychiatric disorder that affects more than 23\u0026nbsp;million individuals worldwide. Over the past century, SCZ has changed little in prevalence and disability\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Cognitive deficit is an independent, core and enduring symptom of SCZ, separate from both positive and negative symptoms, yet closely interrelated with them\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. This deficit contributes to poor clinical outcomes such as functional competence, work skills and an inability to live independently\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Currently, antipsychotics are the primary therapeutic approaches for SCZ in clinical practice\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. However, no significant improvement can be achieved in the treatment of cognitive symptoms with current pharmacotherapy, even though positive symptoms can be significantly alleviated\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Therefore, cognitive deficit is a prevailing issue in the pathogenesis of SCZ, thereby highlighting the paradigm shift from symptom management to functional recovery.\u003c/p\u003e \u003cp\u003eOligodendrocytes (OLs) are myelin-forming cells in the central nervous system. Myelin sheaths wrap axons to promote conduction velocity, provide energy supply, allow neuronal circuits to be finely tuned and synchronized and maintain the proper connectivity among brain regions\u003csup\u003e\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. In the adult brain, cognitive processing highly relies on the interplay of neuronal activity-dependent existing myelin modifications and new myelin formation\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Accumulating evidence indicates a notable loss of OLs and myelin in SCZ pathology. Transcriptome and genetic architecture revealed significant molecular burden in oligodendrocyte lineage cells\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e, and myelin abnormalities have been identified in SCZ patient brains\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Therefore, promoting myelin repair could be available to restore cognitive impairments in SCZ.\u003c/p\u003e \u003cp\u003eIn particular, SCZ psychosis emerges between ages 18\u0026ndash;25, which overlaps with the time for vigorous OLs and myelin development\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. After the first onset, it may persist into adulthood or even old age after multiple relapses. To be mentioned, myelination in the vast majority of brain regions is largely complete by middle age, the new myelin is continuously generated throughout life via the differentiation of adult oligodendroglia precursor cells (OPCs)\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e,\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. The ongoing deposition of OLs and myelin is required for a range of cognitive tasks, including motor skill learning, fear and spatial memory preservation, which is influenced by factors such as neuronal electrical activity and environmental stimuli\u003csup\u003e\u003cspan additionalcitationids=\"CR18\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. However, whether myelin dynamics is influenced and contribute to cognitive deficits during the pathogenesis of SCZ is lack of evidence.\u003c/p\u003e \u003cp\u003eHere, we report a preferential loss of perineuronal OLs (pn-OLs) in MK801-induced mouse model and individuals with SCZ. Strikingly, except for a loss of myelin sheath, we found adult OPC differentiation and new myelin deposition were significantly impaired, which may be due to H3 lysine 4 trimethylation (H3K4me3) deficiency. We confirmed that ablating H3K4 methyltransferase SETD1A in OPCs arrested myelination. While epigenetic rescuing H3K4me3 deficiency repaired pn-OLs and myelin deficits and promote cognitive recovery in MK801 mice. Together, this study identified an epigenetic barrier for OLs generation and myelin production, providing a potential target for myelin repair to improve cognitive impairment.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eHuman postmortem brain tissue and immunostaining\u003c/h2\u003e \u003cp\u003eHuman schizophrenia and healthy comparable post-mortem tissue were provided by the National Health and Disease Human Brain Tissue Resource Center at Zhejiang University in China. The study was performed in accordance with the ethical standards in the Declaration of Helsinki and was approved by the Human Ethics committee of Zhejiang University School of Medicine (Approval Number: 2020\u0026ndash;005). All human tissues were collected following fully informed consent by the donors via a prospective donor scheme. Cases information has been described in Supplementary Table\u0026nbsp;1. The immunohistochemical staining and image capturing have been described previously\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eAnimals\u003c/h3\u003e\n\u003cp\u003eThe NG2-CreERT (The Jackson Laboratory, strain no. 008538) mice were crossed with Tau-mGFP (The Jackson Laboratory, strain no. 021162) mice to obtain NG2-CreERT; Tau-mGFP mice. To specifically delete \u003cem\u003eSetd1a\u003c/em\u003e in OLs, the NG2-CreERT mice were crossed with \u003cem\u003eSetd1a\u003c/em\u003e floxed mice (GemPharmatech Co., Ltd, T013202). Different genotypes of mice were identified using PCR genotyping with the respective primers. The C57BL/6 mice (either sex, 8 weeks old) were obtained from Chongqing Medical Animal Experimentation Center. Animal treatment and study protocols were approved by the Ethical Committee of Chongqing Medical University.\u003c/p\u003e\n\u003ch3\u003eTamoxifen and drug administration\u003c/h3\u003e\n\u003cp\u003eTamoxifen (Sigma-Aldrich, catalog no. T5648) dissolved in ethanol and sunflower oil (1:9, v/v) was administered to induce Cre recombination at a concentration of 30 mg/mL. Adult mice were dosed at 30 mg/kg per day for 4 days. MK801 (MCE, catalog no. HY-15084) was dissolved in saline (0.9% NaCl) at a final concentration of 0.1 mg/mL, and mice were intraperitoneally injected with either the saline vehicle (control group, CTL) or MK801 once daily for 7 consecutive days. After the injection, the mice were allowed to deteriorate for an additional 3 weeks (MK801\u0026thinsp;+\u0026thinsp;1 month group, MK801). MK801 mice were treated with TCP (Sigma, catalog no. P8511, 3 mg/kg/day) and ORY-1001 (Selleck, catalog no. S7795, 10 \u0026micro;g/kg/day) for 7 days while receiving an injection of MK801. Briefly, EdU (Sigma, catalog no. 900584) was dissolved in saline at a concentration of 1 mg/mL, and mice were received a constantly intraperitoneal injection for 10 days, with a dose of 5 mg/kg per day.\u003c/p\u003e\n\u003ch3\u003eBehavior tests\u003c/h3\u003e\n\u003cp\u003eMice were housed in a controlled environment (25℃) with free access to food and water and maintained on a 12-h/12-h light/dark cycle. After each experiment, all the apparatuses were wiped clean with 70% ethanol to prevent a bias due to olfactory cues. For all behavioral experiments, investigators were kept blinded to the treatment groups and mice were gently handled to avoid stress. All of the tests were recorded and analyzed by EthoVision XT 11.5 software and SMART 3.0 software.\u003c/p\u003e\n\u003ch3\u003eY maze\u003c/h3\u003e\n\u003cp\u003eSpontaneous alternative Y-maze test is well-accepted for assessing working memory in mice\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. Briefly, each mouse was gently put in the center of the apparatus (4 \u0026times; 36 cm each arm, arranged at an angle of 120\u0026deg; to each other, with 15.5 cm-high walls), facing one fixed arm, and then allowed to explore freely in the maze for 8 min. During the trial, the number of arm entries and alternations was calculated and measured.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMorris water maze\u003c/h2\u003e \u003cp\u003eThe Morris water maze test was conducted according to a described protocol\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. During the acquisition phase, the water maze pool (depth 60 cm, diameter 120 cm) was divided into four equal quadrants, with a hidden escape platform (diameter 6 cm) fixed in one of the quadrants. Each mouse was allowed to locate the hidden platform for 5 consecutive days from different quadrants (one quadrant per trial). In probe test, the platform was removed, and each mouse was allowed to explore for 1 min, the time and distance traveled in the target quadrant was recorded.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eImmunohistochemistry and EdU staining\u003c/h3\u003e\n\u003cp\u003eImmunofluorescence staining was performed as follows: floating tissue sections (20 \u0026micro;m) were obtained from brains perfused with 4% paraformaldehyde (PFA). The sections were first immersed in 0.5% Triton X-100 for 30 min, followed by blocking with 5% bovine serum albumin (BSA) for 1 h at room temperature, and then incubated with primary antibodies overnight at 4℃. On the following day, tissue sections were incubated with secondary antibodies for 1 h at room temperature. Primary antibodies include: ASPA (1:500, oasis, catalog no. OB-PRT005), MBP (1:500, oasis, catalog no. OB-PRT123), SETD1A (1:200, abcam, catalog no. ab243881), PDGFRα (1:300, R\u0026amp;D, catalog no. AF1062), NeuN (1:500, abcam, catalog no. ab104224), H3K4me3 (1:1000, abcam, catalog no. ab8580), GFP (1:1000, abcam, catalog no. ab5450). EdU staining was strictly performed following the manufacturer\u0026rsquo;s instruction (Beyotime, catalog no. C0081).\u003c/p\u003e\n\u003ch3\u003eWestern blotting\u003c/h3\u003e\n\u003cdiv class=\"Heading\"\u003eWestern blotting\u003c/div\u003e \u003cp\u003eProteins of frozen samples were extracted using RIPA lysis buffer (Beyotime, catalog no. P0013C), which contained protease inhibitors (PMSF, Beyotime, catalog no. ST506). The extracted proteins were then denatured in gel-loading buffer (Beyotime, catalog no. P0286), and 40 \u0026micro;g of protein was separated by 6% and 12% SDS-PAGE gels. The separated proteins were transferred onto PVDF membranes (Millipore, catalog no. IPVH00010), and visualized via enhanced chemiluminescence (ECL) (BIO-OI, catalog no. OI900, China) detection. Band intensity quantification was conducted with Image-Pro Plus software 5.0 (Media Cybernetics). Primary antibodies include: TBP (1:2000, CST, catalog no. 8515), MBP (1:1000), SETD1A (1:5000).\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eRNA extraction and analysis\u003c/h2\u003e \u003cp\u003eThe RNA isolation and reverse transcription were performed as previously described\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. The total RNA was extracted using TRIzol (Invitrogen, catalog no. 15596026), and reverse transcription was conducted following the manufacturer\u0026rsquo;s instruction (Vazyme, catalog no. R233-01). The primers for quantitative RT-PCR (RT-qPCR) are as follows: SETD1A: forward 5\u0026rsquo;-CCCATTAGCAAGAAGGAGAAAGA-3\u0026rsquo;, reverse 5\u0026rsquo;-GCTCAGAAAGCACCCGATTA \u0026minus;\u0026thinsp;3\u0026rsquo;; WDR5: forward 5\u0026rsquo;-GGCCACAGTAACTACGTCTTC-3\u0026rsquo;, reverse 5\u0026rsquo;-CGTCCCATATCCTCACACTTTC-3\u0026rsquo;; KMT2D: forward 5\u0026rsquo;-GTTCGGATAGGAAGGAGCTTATG-3\u0026rsquo;, reverse 5\u0026rsquo;-CATCCTCAAGCCCTGGTAAAT-3\u0026rsquo;; KDM5A: forward 5\u0026rsquo;-CGTGCCTATCACTCTGGATTTA-3\u0026rsquo;, reverse 5\u0026rsquo;-CTGAGCCGTCGATAGTGATTT-3\u0026rsquo;; KDM5B: forward 5\u0026rsquo;-CAAGAGCCCACTGAGAAGAAA-3\u0026rsquo;, reverse 5\u0026rsquo;-TCCACATAAGAGGCACACATAC-3\u0026rsquo;; MBP: forward 5\u0026rsquo;-CTCAGAGGACAGTGATGTGTTT-3\u0026rsquo;, reverse 5\u0026rsquo;-CGCCTTGCCAGTTATTCTTTG-3\u0026rsquo;.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eTransmission electron microscopy\u003c/h2\u003e \u003cp\u003eThe medial prefrontal cortex (mPFC) samples were perfused with 2.5% glutaraldehyde (dissolved in 4% PFA, Macklin, catalog no. G810415). Subsequent detection and capture of these samples were conducted at the Electron Microscopy Center of Chongqing Medical University. The images of these sections were analyzed using ImageJ-Fiji software (version 1.52p).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eSingle-nucleus transcriptome and sequencing\u003c/h2\u003e \u003cp\u003eThe mPFC samples were rapidly frozen in Liquid nitrogen. Subsequent single-nucleus RNA-seq experiment (snRNA-seq) was conducted at the Key Laboratory of Molecular Biology on Infectious Diseases, Chongqing Medical University. a droplet-based microfluidics platform adapted from the inDrop-seq methodology\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. Briefly, single-nucleus suspensions were co-encapsulated with barcoded, degradable hydrogel beads in nanoliter-droplets. Within each droplet, bead degradation released primers to capture polyadenylated mRNA. This was immediately followed by reverse transcription and template switching using a Template Switching Oligo (TSO) to generate barcoded, full-length cDNA. The pooled cDNA was then collected for library preparation.\u003c/p\u003e \u003cp\u003eThe Seurat package (version:5.0.2) was used for cell normalization and regression based on the expression table, according to the UMI counts of each sample and percent of mitochondria rate to obtain the scaled data. Principal component analysis (PCA) was constructed based on the scaled data. The top 2000 high variable genes and top 15 principals were used for uniform manifold approximation and projection (UMAP) construction. Using the graph-based cluster method, we acquired the unsupervised cell cluster result based on the PCA top 15 principals and calculated the marker genes by FindAllMarkers. Significance was defined by the \u003cem\u003ep\u003c/em\u003e-value and false discovery rate (FDR). To identify differentially expressed genes among samples, the function FindMarkers with Wilcox rank sum test algorithm was used under the following criteria: Log2FC\u0026thinsp;\u0026gt;\u0026thinsp;0.25; \u003cem\u003ep\u003c/em\u003e-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05; min.pct\u0026thinsp;\u0026gt;\u0026thinsp;0.1. To enable a systematic analysis of cell-cell communication molecules, cellchat was used to identify cell-to-cell interactions in our dataset. Significant mean and cell communication significance (\u003cem\u003ep\u003c/em\u003e-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05) was calculated based on the interaction and the normalized cell matrix achieved by Seurat normalization.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eImage acquisition and quantification\u003c/h2\u003e \u003cp\u003eFluorescent images were acquired using a confocal laser-scanning microscope (Olympus FV3000 and Nikon NIS-Elements) at appropriate excitation wavelengths. For statistical analysis, the area of the mPFC was measured in each sample for quantification, and was performed using ImageJ-Fiji software (version 1.52p).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eStatistics\u003c/h2\u003e \u003cp\u003eStatistical analyses were performed using GraphPad Prism 9 software. All values were presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. Between-group comparisons were analyzed using two-tailed unpaired \u003cem\u003et\u003c/em\u003e-tests. Data from the acquisition phase of the Morris water maze test was analyzed using repeated-measures two-way ANOVA with separate analyses for each parameter. Each experiment was performed with at least three independent samples. Statistical significance was defined as a \u003cem\u003ep\u003c/em\u003e-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003ePreferential loss of perineuronal oligodendrocytes in MK801-induced mouse model and individuals with schizophrenia\u003c/h2\u003e \u003cp\u003eGlutamatergic hypoactivity, particularly decreased N-Methyl-D-Aspartate (NMDA) receptor function, is one of the most accepted pathophysiological hypotheses for cognitive decline in SCZ\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. To study the pathophysiology of SCZ, we employed MK801-induced mice model, which is a widely-used SCZ model (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). We firstly verified the prolonged effect of MK801 on cognitive function by analyzing the behavioral outcomes. A lower alteration ratio was found in MK801 mice when they spontaneously traveled in the three arms of Y-maze test, although the number of arm entries was comparable (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB), indicating an impairment of spatial memory. Additionally, MK801 mice were trained to locate a hidden platform in a water maze test. Over the 4-day training period, MK801 mice exhibited longer escape latencies, indicating an impaired spatial learning. Moreover, during the probe test, MK801 mice displayed significantly fewer crossings into the target quadrant compared to controls, suggesting an impaired spatial memory (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). To be mentioned, these results may not be due to abnormal locomotor activity, as MK801 mice exhibited similar levels of activity by traveling a similar distance in the probe test (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). Moreover, impaired locomotor activity, depressive and anxious phenotypes were excluded in MK801 mice through different behavioral tests. (Supplementary Fig.\u0026nbsp;1A-D). In addition, social interaction test illustrated that no significant difference was found in social cognition between control and MK801 mice (Supplementary Fig.\u0026nbsp;1E). These results indicate that MK801 model reproduces certain aspects of cognitive dysfunction associated with schizophrenia.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo verify the alterations of oligodendroglia lineage cells, we analyzed the cell density and found that ASPA\u003csup\u003e+\u003c/sup\u003e mature OLs (MOLs) instead of PDGFRα\u003csup\u003e+\u003c/sup\u003e OPCs were decreased in mPFC of MK801 mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD-E). In this study, the relative positional relationship between pn-OLs and neurons was validated through immunostaining and 3D reconstruction (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eF). Electron microscopy analysis further confirmed the attachment of pn-OL could form long curved or concave impressions on the contacting neuron (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eG). We confirmed that 29.2% of ASPA\u003csup\u003e+\u003c/sup\u003e MOLs are pn-OLs in mPFC of adult mice at the age of 3 months (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eH). More strikingly, we found a significant decrease of pn-OLs rather than non-pn-OLs in the mPFC of MK801 mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eI). To further validate that the loss of pn-OLs is a prominent feature in SCZ, we examined the density of Olig2\u003csup\u003e+\u003c/sup\u003e oligodendroglia lineage cells in the cortex of SCZ patients and age-matched controls. We identified a preferential loss of Olig2\u003csup\u003e+\u003c/sup\u003e pn-OLs in cortex from SCZ patients (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eJ).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eExtensive myelin loss and arrested new myelin deposition in MK801 mice\u003c/h2\u003e \u003cp\u003eIn fact, the morphology of pn-OLs shows no substantial difference from that of typical myelinating OLs\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Thus, we questioned whether loss of pn-OLs could result in myelin deficits. We found that MBP labeled myelin fibers significantly decreased in mPFC of MK801 mice as compared to control mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). To access more details of myelin ultrastructure, we analyzed the organization of myelin sheaths using a transmission electron microscope. Consistently, we found the myelin sheaths in the mPFC of MK801 mice were thinner than those in control mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). It is important to note that the densities of NeuN-positive neurons and NF200-positive axons were not altered in the brains after MK801 treatment (Supplementary Fig.\u0026nbsp;2A-B). We examined axonal degeneration by labeling for SMI-32P, and axonal degeneration was not apparent in the mPFC of MK801 mice (Supplementary Fig.\u0026nbsp;2B). These results suggest that MK801 treatment led to a loss of pn-OLs and robust myelin deficiency without causing evident neuronal degeneration.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo directly assess myelin generation capacity in mPFC, we further employed the NG2-CreERT; Tau-mGFP mouse line to label newly formed myelin sheaths\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. As the expression level of Tau is high in MOLs but low in OPCs, membrane-bound GFP (mGFP) accumulates only in newly-differentiated OLs and the newly-formed myelin sheaths formed after tamoxifen administration (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). Notably, abundant mGFP-positive myelin sheaths were observed in the mPFC of adult mice 28 days post-induction, indicating robust new myelin deposition within one month. In contrast, mGFP-positive sheaths were sparse in the mPFC of MK801 mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD). In addition, we found the generation of mGFP\u003csup\u003e+\u003c/sup\u003e new pn-OLs was significantly impaired in mPFC of MK801 mice compared to control mice. Moreover, newly-generated pn-OLs exhibited a lack of myelin production capacity after MK801 treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eE). These results confirmed that the generation of pn-OLs and new myelin were significantly arrested in MK801 mice.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eAdult oligodendroglia precursor cells differentiation was impaired in MK801 mice\u003c/h2\u003e \u003cp\u003eMK801 treatment is associated with augmented glutamate release in the mPFC, resembling findings in first-episode schizophrenia\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. In our study, we also observed an increase in synaptic vesicles within presynaptic bouton of excitatory glutamatergic neuron in mPFC of MK801 mice (Supplementary Fig.\u0026nbsp;3). To delve deeper into the potential molecular alterations affected by MK801, we profiled 40 089 nuclei from 6 mPFC tissues. Unsupervised clustering of the transcriptomic data revealed distinct populations of all major brain cell types (Supplementary Fig.\u0026nbsp;4A). Those clusters were annotated as excitatory neurons (\u003cem\u003eCamk2a\u003c/em\u003e, \u003cem\u003eSlc17a7\u003c/em\u003e and \u003cem\u003eSatb2\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e), inhibitory neurons (\u003cem\u003ePvalb\u003c/em\u003e, \u003cem\u003eVip\u003c/em\u003e, \u003cem\u003eGad1\u003c/em\u003e and \u003cem\u003eGad2\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e), astrocytes (\u003cem\u003eAldh1l1\u003c/em\u003e and \u003cem\u003eAqp4\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e), OLs (\u003cem\u003ePlp1\u003c/em\u003e, \u003cem\u003eMbp\u003c/em\u003e, \u003cem\u003eMobp\u003c/em\u003e, \u003cem\u003ePdgfrα\u003c/em\u003e, \u003cem\u003eCspg4\u003c/em\u003e and \u003cem\u003eGpr17\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e), endothelial cells (\u003cem\u003eRgs5\u003c/em\u003e and \u003cem\u003eCldn5\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e), and microglia (\u003cem\u003eCx3cr1\u003c/em\u003e, \u003cem\u003eC1qb\u003c/em\u003e and \u003cem\u003eTmem119\u003c/em\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e) (Supplementary Fig.\u0026nbsp;4B). We then compared single-nuclei transcriptional profiles of 6 clusters including oligodendroglia lineage cells from the MK801 and CTL mice (Supplementary Fig.\u0026nbsp;4C, Supplementary Table\u0026nbsp;2). Cluster analysis of OLs further revealed three subclusters: OPCs, newly formed OLs (NFOLs), and MOLs, which were based on those established markers including \u003cem\u003ePdgfrα\u003c/em\u003e, \u003cem\u003eCspg4\u003c/em\u003e, \u003cem\u003eGpr17\u003c/em\u003e, \u003cem\u003eEnpp6\u003c/em\u003e, \u003cem\u003eMbp\u003c/em\u003e, and \u003cem\u003ePlp1\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA-C) \u003csup\u003e30, 31\u003c/sup\u003e. Numerous differentially expressed genes (DEGs) were observed across OL subclusters in MK801-treated versus control mice with a pronounced downregulation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD, Supplementary Table\u0026nbsp;3). UMAP visualization and cellular proportion analysis demonstrated a significant increase of OPCs and a decrease of MOLs in the MK801-treated group compared to controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE), implying an impaired OPCs differentiation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003ePrevious studies have found that functional synapses are formed between OPCs and glutamatergic neurons, enabling OPCs to perceive and decode neuronal activity\u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e. We applied a predictive tool to explore the relationship of ligands of excitatory neurons to the receptors expressed in OL subclusters. We found that MK801 treatment disrupted communication mediated by the glutamate signaling between OPCs, NFOLs and excitatory neurons (Supplementary Fig.\u0026nbsp;4D). Consistently, GO analyses of downregulated genes in OPCs and NFOLs further proved that synapse related cellular components were severely affected in MK801 mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF-H). To investigate the influence of aberrant glutamate signaling on OPCs differentiation, we subsequently traced the dividing OPCs using EdU and found the number of ASPA\u003csup\u003e+\u003c/sup\u003e/EdU\u003csup\u003e+\u003c/sup\u003e newly differentiated OLs was significantly decreased in the mPFC of MK801 mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eI). These results suggest MK801-induced aberrant glutamate signaling may impair adult OPCs differentiation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eNewly differentiated oligodendrocytes are epigenetically silenced via H3K4me3 insufficiency\u003c/h2\u003e \u003cp\u003eThe acute administration of MK801 causes long-term differentiation barriers in adult OPCs, which implies an endogenous epigenetic abnormality in OLs. In fact, chromatin compaction positively correlates with the maturation stage of OLs\u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. Here we confirmed that euchromatin marked by H3K4me3 (a transcriptionally active chromatin modification), was higher in OPCs and significantly decreased in mature stage (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA-B). We found MOLs within the mPFC of MK801 mice exhibited aberrantly lower or undetectable levels of H3K4me3 compared to that of MOLs in control mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC). Notably, the newly-generated MOLs expressed higher levels of H3K4me3 than pre-existing MOLs, implying a higher transcriptional activity. However, a global loss of the active H3K4me3 modification was observed in newly-formed MOLs after MK801 treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD), suggesting an epigenetic suppression in these cells. We further examined the enzymes regulating H3K4me3 status. Strikingly, SETD1A, a key H3K4 methyltransferase, was significantly downregulated in MK801 mice, despite no change in the expression of its functional partners (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE). In contrast, the lysine demethylases KDM5A and KDM5B were significantly upregulated (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF). These results suggest that the H3K4me3 deficiency may set a barrier inhibiting OLs produce myelin in MK801 mice.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo confirm the role of H3K4me3 in OLs myelination, we analyzed the single-cell RNA sequencing (scRNA-seq) data of mPFC cells that were freshly isolated from young-adult \u003cem\u003eSetd1a\u003c/em\u003e knockout mice\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Using a Seurat-based workflow\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e, 3 main clusters of oligodendroglia lineage cells were successfully identified (Supplementary Fig.\u0026nbsp;5A), including OPCs, NFOLs and MOLs, based on cluster marker genes and known cell type markers (Supplementary Fig.\u0026nbsp;5B)\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. Cell proportion analysis revealed a decreasing trend of NFOLs in mPFC from \u003cem\u003eSetd1a\u003c/em\u003e\u003csup\u003e\u003cem\u003e+/\u0026minus;\u003c/em\u003e\u003c/sup\u003e heterozygous mouse (Supplementary Fig.\u0026nbsp;5C). GO analyses of differentially expressed genes further proved that myelin sheath related cellular components were severely affected by \u003cem\u003eSetd1a\u003c/em\u003e deficiency (Supplementary Fig.\u0026nbsp;5D). To validate the impact of H3K4me3 insufficiency on myelination \u003cem\u003ein vivo\u003c/em\u003e, we deleted \u003cem\u003eSetd1a\u003c/em\u003e in the OPCs (\u003cem\u003eSetd1a\u003c/em\u003e cKO) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA). The knockout efficiency of \u003cem\u003eSetd1a\u003c/em\u003e was confirmed by the diminished expression of the SETD1A protein in corpus callosum of \u003cem\u003eSetd1a\u003c/em\u003e cKO mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB). Strikingly, both the transcript and protein levels of MBP, a key component of myelin, significantly reduced in \u003cem\u003eSetd1a\u003c/em\u003e cKO mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB-C). As expected, SETD1A deficiency led to widespread decrease of H3K4me3 level in OPCs (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD). To further validate that myelin deficiency is a prominent feature, we examined myelin patterns in the mPFC. The areas of MBP\u003csup\u003e+\u003c/sup\u003e myelinated fibers significantly decreased in mPFC of \u003cem\u003eSetd1a\u003c/em\u003e cKO mice compared to wildtype littermates (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE). While the disturbed myelin formation in \u003cem\u003eSetd1a\u003c/em\u003e cKO mice was further validated via electron microscopy (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF). These results suggest that H3K4me3 status is essential for OLs myelination.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003ePharmacological strategy counteracting H3K4me3 deficiency restoring myelin deficits and cognitive dysfunction in MK801 mice.\u003c/b\u003e \u003c/p\u003e \u003cp\u003eGiven the therapeutic implications of these results, we investigated whether counteracting H3K4me3 deficiency would enable OL myelin-producing programs to proceed and therefore improve cognition in MK801 mice. A previous study has proved that lysine specific demethylase 1 (LSD1), a demethylase that counteracts SETD1A, is the most abundant demethylase in the frontal cortex \u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e. Building on this finding, we examined the effects of inhibiting LSD1 on myelin deficits with systemic administration of TCP and ORY-1001 for 1 month (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA). As expected, the drug treatment significantly increased the H3K4me3 level in EdU\u003csup\u003e+\u003c/sup\u003e newly-generated MOLs, as compared with the vehicle group (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB). To be noted, the density of pn-OLs was recovered after those drug treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC). More strikingly, immunostaining showed that the deficiency of MBP\u003csup\u003e+\u003c/sup\u003e myelinated fibers in the mPFC of MK801 mice was repaired after drug treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eD). Electron microscopy further confirmed increased myelin thickness in the mPFC of drug-treated mice compared to vehicle controls (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE). These results suggest that rescuing H3K4me3 deficiency in OLs enhances the generation of pn-OLs and myelin repair in the MK801 brains.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eWe next sought to determine whether \u003cem\u003ein vivo\u003c/em\u003e post-developmental LSD1 antagonism by TCP could repair cognitive deficits parallel to enhanced myelination. Mice were subjected to the Y-maze and Morris water maze tasks. MK801 mice treated with TCP and ORY-1001 showed a higher alteration ratio during the Y-maze task, indicating improved working memory compared to vehicle-treated mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eF). Moreover, over 4 days of training, all groups required progressively less time to reach the platform, reflecting a similar degree of spatial learning. While during the probe test, in which the hidden platform was removed, the drug-treated mice spent more time and traveled a longer distance in the quadrant that previously contained the platform relative to vehicle mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eG). Thus, systemic treatment with a H3K4me3 targeting drug enhancing myelin repair could improve cognitive performance in MK801 mice.\u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eIn this study, we demonstrated a preferential loss of pn-OLs in both SCZ patients and MK801 mice. In addition to widespread myelin loss, we provided the first evidence that endogenous OL and myelin renewal were severely blocked after an acute MK801 treatment, which contributes to cognitive deficits in mice. Moreover, we identified H3K4me3 as a crucial factor governing new myelin deposition. Finally, we proved that pharmacologically rescuing H3K4me3 deficiency significantly enhanced myelin repair, and thereafter improved the cognitive performance of MK801 mice.\u003c/p\u003e \u003cp\u003eCognitive decline may begin in adolescents diagnosed with SCZ, even precede illness onset, and accelerate in adulthood\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. Supporting this notion, we employed a mouse model by injecting MK801 in young-adult mice. In fact, the MK801 model was proved to have good performance, construct and predictive validity for studying the pathophysiology of cognitive dysfunction in SCZ\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e. Previous study found that MK801-induced disruption of NMDA receptor function during adolescence can lead to persistent dysfunction of the prefrontal cortex in adulthood\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. Similarly, we found that an acute MK801 treatment in young-adult mice led to prolonged cognitive deficits, especially the working memory and spatial memory (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). However, in our study, social cognition as well as emotional alterations was intact in MK801 mice (Supplementary Fig.\u0026nbsp;1), which highlights the heterogeneity of cognitive impairments in patients. In fact, cognitive heterogeneity not only compromises the prediction of clinical outcomes but also impedes the development of precision treatment strategies.\u003c/p\u003e \u003cp\u003eAccumulating evidence indicates that both OLs and myelin are significantly altered during the pathogenesis of schizophrenia. In this study, we clearly identified a preferential loss of pn-OLs in the cortex of SCZ patients as well as the MK801 mouse model (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The term of pn-OLs was coined by the pioneering neurohistologist P\u0026iacute;o del R\u0026iacute;o Hortega decades ago\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e, while the function of these cells in the cortex remains unknown. We confirmed that pn-OLs were able to produce myelin, and deficits of adult OPC differentiation and new myelin formation led to myelin deficiency under MK801 treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). It is known that neuronal activity can affect OPCs differentiation, maturation and myelin formation through the glutamate signaling \u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e. In this study, we found that MK801 treatment disrupted the communication between OPCs/NFOLs and excitatory neurons, which may impair OPCs differentiation (Supplementary Fig.\u0026nbsp;4). Because our understanding of the functional interplay between OLs and excitatory neurons is still at its infancy, the precise mechanism governing pn-OLs function still remains undefined. Besides myelination, accumulating evidence suggests that pn-OLs can provide metabolic and neurotrophic support to nearby neurons\u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e. Interestingly, studies have shown that the localization of pn-OLs exhibits neuronal selectivity, preferentially associating with glutamatergic neurons compared to GABAergic interneurons\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. What\u0026rsquo;s more, the pn-OLs express ionotropic and metabotropic glutamate receptors, glutamate transporters and glutamine synthetase (GS) for converting glutamate to glutamine\u003csup\u003e\u003cspan additionalcitationids=\"CR45 CR46\" citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e. These findings further support that pn-OLs appear to be sensitive to glutamate metabolism, which may in turn facilitate the metabolism of neurons and modulate their activity\u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eImpaired adult OPCs differentiation has long been considered the primary cause of remyelination failure\u003csup\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e. In other cases, even though MOLs are present, they fail to generate new myelin sheaths, which may due to epigenetic barrier\u003csup\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e. From the insight of development, epigenetic regulation is crucial for OPCs differentiation\u003csup\u003e\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e. Here, after MK801 insult, we found a global loss of the active H3K4me3 modification in the newly-generated MOLs (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Impairment of the epigenetic barrier mediated by H3K4me3 compromised the ability of MOLs to produce myelin. This process could be primarily mediated through a series of enzymatic regulation, such as SETD1A, KDM5A and KDM5B (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). To be noted, \u003cem\u003eSetd1a\u003c/em\u003e has recently been identified as a risk gene for SCZ, and \u003cem\u003eSetd1a\u003c/em\u003e mutations confer a large increase in disease risk\u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e. Recently, multiple studies have elucidated how \u003cem\u003eSetd1a\u003c/em\u003e affects neuronal development and synaptic function, and thereafter contribute to the onset of SCZ\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e. However, whether \u003cem\u003eSetd1a\u003c/em\u003e affects OLs function in SCZ remains unclear. In this study, we provide the first evidence that SETD1A, a key regulator of H3K4me3, is tightly related to OL myelination (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e), shedding light on H3K4me3 status in white matter etiology of SCZ.\u003c/p\u003e \u003cp\u003eNumerous studies have explored strategies to enhance myelin formation in disease treatment. Chen, Jing-Fei et al. found that promoting myelinogenesis significantly improved performance in memory-related tasks and alleviated cognitive impairment in a mouse model of Alzheimer's disease\u003csup\u003e\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. These findings suggest that myelin repair represents a potentially effective strategy for ameliorating cognitive deficits in SCZ. Recently, a number of histone demethylase inhibitors with good blood-brain barrier permeability have been identified. We speculate that such compounds are likely to induce substantial changes in methylation states, thereby counteracting the effects of H3K4me3 deficiency. In this study, we found treatment with two LSD1 inhibitors, TCP and ORY-1001, rescued pn-OLs and myelin deficits and ameliorated cognitive deficits in MK801-treated mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e), This is an exciting observation, as it presents a promising target for promoting endogenous remyelination, and thereafter overcome myelin related cognitive deficits in major psychiatric disorders represented by SCZ.\u003c/p\u003e \u003cp\u003eIn conclusion, our findings identified the pathological potential of pn-OLs, and provided new evidence of myelin generation deficits in SCZ, possibly due to an H3K4me3-mediated epigenetic barrier. Targeting the H3K4me3 status in OLs could be a novel therapeutic strategy for cognitive deficits in SCZ.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eDATA AVAILABILITY\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data are available in the main text and supplementary materials. The single-nucleus transcriptome statistics are available from the corresponding author, XC, upon reasonable request.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the National Natural Science Foundation of China (32130019, 32000684), the National Key Research and Development Program of China (2021YFA1100203) and Foundation of Chongqing Educational Committee (KJQN202500421).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eAUTHOR CONTRIBUTIONS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization: XC, BW, and LZ; Methodology: XC and YC; Investigation: XC, YC, YC, ML, MZ, MJ, SL, BT, CZ, XY, AG, QJ, WX, JL, QL, YM, JX, JN and LX; Visualization: XC, YC, ML and YC; Funding acquisition: XC and BW; Project administration: XC and BW; Supervision: XC and BW; Writing\u0026ndash;original draft: XC, YC, ML, YC, QL and MJ; Writing\u0026ndash;review \u0026amp; editing: XC, YC, ML, YC, QL and MJ.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eCOMPETING INTERESTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eHegarty JD, Baldessarini RJ, Tohen M, Waternaux C, Oepen G. One hundred years of schizophrenia: a meta-analysis of the outcome literature. \u003cem\u003eThe American journal of psychiatry\u003c/em\u003e 1994; 151(10): 1409\u0026ndash;1416.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRodriguez-Jimenez R, Bagney A, Mezquita L, Martinez-Gras I, Sanchez-Morla EM, Mesa N \u003cem\u003eet al.\u003c/em\u003e Cognition and the five-factor model of the positive and negative syndrome scale in schizophrenia. \u003cem\u003eSchizophrenia research\u003c/em\u003e 2013; 143(1): 77\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBowie CR, Leung WW, Reichenberg A, McClure MM, Patterson TL, Heaton RK \u003cem\u003eet al.\u003c/em\u003e Predicting schizophrenia patients' real-world behavior with specific neuropsychological and functional capacity measures. \u003cem\u003eBiological psychiatry\u003c/em\u003e 2008; 63(5): 505\u0026ndash;511.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKane JM, Lencz T. Cognitive deficits in schizophrenia: short-term and long-term. \u003cem\u003eWorld psychiatry: official journal of the World Psychiatric Association (WPA)\u003c/em\u003e 2008; 7(1): 29\u0026ndash;30.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEfthimiou O, Taipale H, Radua J, Schneider-Thoma J, Pinz\u0026oacute;n-Espinosa J, Ortu\u0026ntilde;o M \u003cem\u003eet al.\u003c/em\u003e Efficacy and effectiveness of antipsychotics in schizophrenia: network meta-analyses combining evidence from randomised controlled trials and real-world data. \u003cem\u003eThe lancet Psychiatry\u003c/em\u003e 2024; 11(2): 102\u0026ndash;111.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMonteleone P, Cascino G, Monteleone AM, Rocca P, Rossi A, Bertolino A \u003cem\u003eet al.\u003c/em\u003e Prevalence of antipsychotic-induced extrapyramidal symptoms and their association with neurocognition and social cognition in outpatients with schizophrenia in the \"real-life\". \u003cem\u003eProgress in neuro-psychopharmacology \u0026amp; biological psychiatry\u003c/em\u003e 2021; 109: 110250.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eF\u0026uuml;nfschilling U, Supplie LM, Mahad D, Boretius S, Saab AS, Edgar J \u003cem\u003eet al.\u003c/em\u003e Glycolytic oligodendrocytes maintain myelin and long-term axonal integrity. \u003cem\u003eNature\u003c/em\u003e 2012; 485(7399): 517\u0026ndash;521.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee Y, Morrison BM, Li Y, Lengacher S, Farah MH, Hoffman PN \u003cem\u003eet al.\u003c/em\u003e Oligodendroglia metabolically support axons and contribute to neurodegeneration. \u003cem\u003eNature\u003c/em\u003e 2012; 487(7408): 443\u0026ndash;448.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHunt BA, Tewarie PK, Mougin OE, Geades N, Jones DK, Singh KD \u003cem\u003eet al.\u003c/em\u003e Relationships between cortical myeloarchitecture and electrophysiological networks. \u003cem\u003eProceedings of the National Academy of Sciences of the United States of America\u003c/em\u003e 2016; 113(47): 13510\u0026ndash;13515.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunyeshyaka M, Fields RD. Oligodendroglia are emerging players in several forms of learning and memory. \u003cem\u003eCommunications biology\u003c/em\u003e 2022; 5(1): 1148.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBacmeister CM, Huang R, Osso LA, Thornton MA, Conant L, Chavez AR \u003cem\u003eet al.\u003c/em\u003e Motor learning drives dynamic patterns of intermittent myelination on learning-activated axons. \u003cem\u003eNature neuroscience\u003c/em\u003e 2022; 25(10): 1300\u0026ndash;1313.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMartins-de-Souza D. Proteome and transcriptome analysis suggests oligodendrocyte dysfunction in schizophrenia. \u003cem\u003eJournal of psychiatric research\u003c/em\u003e 2010; 44(3): 149\u0026ndash;156.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBiological insights from 108 schizophrenia-associated genetic loci. \u003cem\u003eNature\u003c/em\u003e 2014; 511(7510): 421\u0026ndash;427.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFlynn SW, Lang DJ, Mackay AL, Goghari V, Vavasour IM, Whittall KP \u003cem\u003eet al.\u003c/em\u003e Abnormalities of myelination in schizophrenia detected in vivo with MRI, and post-mortem with analysis of oligodendrocyte proteins. \u003cem\u003eMolecular psychiatry\u003c/em\u003e 2003; 8(9): 811\u0026ndash;820.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eInsel TR. Rethinking schizophrenia. \u003cem\u003eNature\u003c/em\u003e 2010; 468(7321): 187\u0026ndash;193.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHill RA, Li AM, Grutzendler J. Lifelong cortical myelin plasticity and age-related degeneration in the live mammalian brain. \u003cem\u003eNature neuroscience\u003c/em\u003e 2018; 21(5): 683\u0026ndash;695.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHughes EG, Orthmann-Murphy JL, Langseth AJ, Bergles DE. Myelin remodeling through experience-dependent oligodendrogenesis in the adult somatosensory cortex. \u003cem\u003eNature neuroscience\u003c/em\u003e 2018; 21(5): 696\u0026ndash;706.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePan S, Mayoral SR, Choi HS, Chan JR, Kheirbek MA. Preservation of a remote fear memory requires new myelin formation. \u003cem\u003eNature neuroscience\u003c/em\u003e 2020; 23(4): 487\u0026ndash;499.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBacmeister CM, Barr HJ, McClain CR, Thornton MA, Nettles D, Welle CG \u003cem\u003eet al.\u003c/em\u003e Motor learning promotes remyelination via new and surviving oligodendrocytes. \u003cem\u003eNature neuroscience\u003c/em\u003e 2020; 23(7): 819\u0026ndash;831.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYu G, Su Y, Guo C, Yi C, Yu B, Chen H \u003cem\u003eet al.\u003c/em\u003e Pathological oligodendrocyte precursor cells revealed in human schizophrenic brains and trigger schizophrenia-like behaviors and synaptic defects in genetic animal model. \u003cem\u003eMolecular psychiatry\u003c/em\u003e 2022; 27(12): 5154\u0026ndash;5166.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoi YK, Maki T, Mandeville ET, Koh SH, Hayakawa K, Arai K \u003cem\u003eet al.\u003c/em\u003e Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury. \u003cem\u003eNature medicine\u003c/em\u003e 2016; 22(11): 1335\u0026ndash;1341.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVorhees CV, Williams MT. Morris water maze: procedures for assessing spatial and related forms of learning and memory. \u003cem\u003eNature protocols\u003c/em\u003e 2006; 1(2): 848\u0026ndash;858.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen X, Ku L, Mei R, Liu G, Xu C, Wen Z \u003cem\u003eet al.\u003c/em\u003e Novel schizophrenia risk factor pathways regulate FEZ1 to advance oligodendroglia development. \u003cem\u003eTranslational psychiatry\u003c/em\u003e 2017; 7(12): 1293.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKlein AM, Mazutis L, Akartuna I, Tallapragada N, Veres A, Li V \u003cem\u003eet al.\u003c/em\u003e Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. \u003cem\u003eCell\u003c/em\u003e 2015; 161(5): 1187\u0026ndash;1201.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuck SA, Quincy Erickson-Oberg M, Logan RW, Freyberg Z. Relevance of interactions between dopamine and glutamate neurotransmission in schizophrenia. \u003cem\u003eMolecular psychiatry\u003c/em\u003e 2022; 27(9): 3583\u0026ndash;3591.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBernstein HG, Keilhoff G, Dobrowolny H, Guest PC, Steiner J. Perineuronal oligodendrocytes in health and disease: the journey so far. \u003cem\u003eReviews in the neurosciences\u003c/em\u003e 2019; 31(1): 89\u0026ndash;99.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakasaki C, Yamasaki M, Uchigashima M, Konno K, Yanagawa Y, Watanabe M. Cytochemical and cytological properties of perineuronal oligodendrocytes in the mouse cortex. \u003cem\u003eThe European journal of neuroscience\u003c/em\u003e 2010; 32(8): 1326\u0026ndash;1336.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang F, Ren SY, Chen JF, Liu K, Li RX, Li ZF \u003cem\u003eet al.\u003c/em\u003e Myelin degeneration and diminished myelin renewal contribute to age-related deficits in memory. \u003cem\u003eNature neuroscience\u003c/em\u003e 2020; 23(4): 481\u0026ndash;486.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZou Y, Zhang H, Chen X, Ji W, Mao L, Lei H. Age-dependent effects of (+)-MK801 treatment on glutamate release and metabolism in the rat medial prefrontal cortex. \u003cem\u003eNeurochemistry international\u003c/em\u003e 2019; 129: 104503.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHou J, Zhou Y, Cai Z, Terekhova M, Swain A, Andhey PS \u003cem\u003eet al.\u003c/em\u003e Transcriptomic atlas and interaction networks of brain cells in mouse CNS demyelination and remyelination. \u003cem\u003eCell reports\u003c/em\u003e 2023; 42(4): 112293.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDennis DJ, Wang BS, Karamboulas K, Kaplan DR, Miller FD. Single-cell approaches define two groups of mammalian oligodendrocyte precursor cells and their evolution over developmental time. \u003cem\u003eStem cell reports\u003c/em\u003e 2024; 19(5): 654\u0026ndash;672.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBergles DE, Roberts JD, Somogyi P, Jahr CE. Glutamatergic synapses on oligodendrocyte precursor cells in the hippocampus. \u003cem\u003eNature\u003c/em\u003e 2000; 405(6783): 187\u0026ndash;191.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu J, Magri L, Zhang F, Marsh NO, Albrecht S, Huynh JL \u003cem\u003eet al.\u003c/em\u003e Chromatin landscape defined by repressive histone methylation during oligodendrocyte differentiation. \u003cem\u003eThe Journal of neuroscience: the official journal of the Society for Neuroscience\u003c/em\u003e 2015; 35(1): 352\u0026ndash;365.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen R, Liu Y, Djekidel MN, Chen W, Bhattacherjee A, Chen Z \u003cem\u003eet al.\u003c/em\u003e Cell type-specific mechanism of Setd1a heterozygosity in schizophrenia pathogenesis. \u003cem\u003eSci Adv\u003c/em\u003e 2022; 8(9): eabm1077.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eButler A, Hoffman P, Smibert P, Papalexi E, Satija R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. \u003cem\u003eNature biotechnology\u003c/em\u003e 2018; 36(5): 411\u0026ndash;420.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMukai J, Cannav\u0026ograve; E, Crabtree GW, Sun Z, Diamantopoulou A, Thakur P \u003cem\u003eet al.\u003c/em\u003e Recapitulation and Reversal of Schizophrenia-Related Phenotypes in Setd1a-Deficient Mice. \u003cem\u003eNeuron\u003c/em\u003e 2019; 104(3): 471\u0026ndash;487.e412.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee M, Cernvall M, Borg J, Plav\u0026eacute;n-Sigray P, Larsson C, Erhardt S \u003cem\u003eet al.\u003c/em\u003e Cognitive Function and Variability in Antipsychotic Drug-Naive Patients With First-Episode Psychosis: A Systematic Review and Meta-Analysis. \u003cem\u003eJAMA psychiatry\u003c/em\u003e 2024; 81(5): 468\u0026ndash;476.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJonas K, Lian W, Callahan J, Ruggero CJ, Clouston S, Reichenberg A \u003cem\u003eet al.\u003c/em\u003e The Course of General Cognitive Ability in Individuals With Psychotic Disorders. \u003cem\u003eJAMA psychiatry\u003c/em\u003e 2022; 79(7): 659\u0026ndash;666.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown JW, Rueter LE, Zhang M. Predictive validity of a MK-801-induced cognitive impairment model in mice: implications on the potential limitations and challenges of modeling cognitive impairment associated with schizophrenia preclinically. \u003cem\u003eProgress in neuro-psychopharmacology \u0026amp; biological psychiatry\u003c/em\u003e 2014; 49: 53\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFlores-Barrera E, Thomases DR, Tseng KY. MK-801 Exposure during Adolescence Elicits Enduring Disruption of Prefrontal E-I Balance and Its Control of Fear Extinction Behavior. \u003cem\u003eThe Journal of neuroscience: the official journal of the Society for Neuroscience\u003c/em\u003e 2020; 40(25): 4881\u0026ndash;4887.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi C, Xiao L, Liu X, Yang W, Shen W, Hu C \u003cem\u003eet al.\u003c/em\u003e A functional role of NMDA receptor in regulating the differentiation of oligodendrocyte precursor cells and remyelination. \u003cem\u003eGlia\u003c/em\u003e 2013; 61(5): 732\u0026ndash;749.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGautier HO, Evans KA, Volbracht K, James R, Sitnikov S, Lundgaard I \u003cem\u003eet al.\u003c/em\u003e Neuronal activity regulates remyelination via glutamate signalling to oligodendrocyte progenitors. \u003cem\u003eNature communications\u003c/em\u003e 2015; 6: 8518.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDu Y, Dreyfus CF. Oligodendrocytes as providers of growth factors. \u003cem\u003eJournal of neuroscience research\u003c/em\u003e 2002; 68(6): 647\u0026ndash;654.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eK\u0026aacute;rad\u0026oacute;ttir R, Cavelier P, Bergersen LH, Attwell D. NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. \u003cem\u003eNature\u003c/em\u003e 2005; 438(7071): 1162\u0026ndash;1166.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeSilva TM, Kabakov AY, Goldhoff PE, Volpe JJ, Rosenberg PA. Regulation of glutamate transport in developing rat oligodendrocytes. \u003cem\u003eThe Journal of neuroscience: the official journal of the Society for Neuroscience\u003c/em\u003e 2009; 29(24): 7898\u0026ndash;7908.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYamazaki Y, Hozumi Y, Kaneko K, Fujii S. Modulatory Effects of Perineuronal Oligodendrocytes on Neuronal Activity in the Rat Hippocampus. \u003cem\u003eNeurochemical research\u003c/em\u003e 2018; 43(1): 27\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD'Amelio F, Eng LF, Gibbs MA. Glutamine synthetase immunoreactivity is present in oligodendroglia of various regions of the central nervous system. \u003cem\u003eGlia\u003c/em\u003e 1990; 3(5): 335\u0026ndash;341.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKuhlmann T, Miron V, Cui Q, Wegner C, Antel J, Br\u0026uuml;ck W. Differentiation block of oligodendroglial progenitor cells as a cause for remyelination failure in chronic multiple sclerosis. \u003cem\u003eBrain: a journal of neurology\u003c/em\u003e 2008; 131(Pt 7): 1749\u0026ndash;1758.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu X, Xin DE, Zhong X, Zhao C, Li Z, Zhang L \u003cem\u003eet al.\u003c/em\u003e Small-molecule-induced epigenetic rejuvenation promotes SREBP condensation and overcomes barriers to CNS myelin regeneration. \u003cem\u003eCell\u003c/em\u003e 2024; 187(10): 2465\u0026ndash;2484.e2422.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEmery B, Lu QR. Transcriptional and Epigenetic Regulation of Oligodendrocyte Development and Myelination in the Central Nervous System. \u003cem\u003eCold Spring Harbor perspectives in biology\u003c/em\u003e 2015; 7(9): a020461.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh T, Kurki MI, Curtis D, Purcell SM, Crooks L, McRae J \u003cem\u003eet al.\u003c/em\u003e Rare loss-of-function variants in SETD1A are associated with schizophrenia and developmental disorders. \u003cem\u003eNature neuroscience\u003c/em\u003e 2016; 19(4): 571\u0026ndash;577.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSu X, Zhang H, Hong Y, Yang Q, Wang L, Le T \u003cem\u003eet al.\u003c/em\u003e Mutations of schizophrenia risk gene SETD1A dysregulate synaptic function in human neurons. \u003cem\u003eMolecular psychiatry\u003c/em\u003e 2025.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen JF, Liu K, Hu B, Li RR, Xin W, Chen H \u003cem\u003eet al.\u003c/em\u003e Enhancing myelin renewal reverses cognitive dysfunction in a murine model of Alzheimer's disease. \u003cem\u003eNeuron\u003c/em\u003e 2021; 109(14): 2292\u0026ndash;2307.e2295.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"molecular-psychiatry","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"mp","sideBox":"Learn more about [Molecular Psychiatry](http://www.nature.com/mp/)","snPcode":"41380","submissionUrl":"https://mts-mp.nature.com/cgi-bin/main.plex","title":"Molecular Psychiatry","twitterHandle":"@molpsychiatry","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8000305/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8000305/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCognitive deficits are core symptoms of schizophrenia (SCZ) and major contributors to disability. Beyond gray matter loss, significant oligodendrocytes (OLs) and myelin pathologies have been well-identified in SCZ patients. Cognitive processing highly relies on the myelination in the adult brain. However, whether and how OLs and myelin dynamics contribute to disease pathology remains unexplored. Here, we characterized a preferential loss of perineuronal OLs in both the dizocilpine (MK801)-induced mouse model and postmortem SCZ brain tissue. The extent of myelination was decreased in the medial prefrontal cortex (mPFC) of MK801 mice. Strikingly, spontaneous myelin renewal was suppressed due to a global deficiency in H3 lysine 4 trimethylation (H3K4me3) in OLs. Conditional knockout of Setd1a, the specific H3K4 methyltransferase in oligodendroglia precursor cells (OPCs), impaired myelination. Notably, pharmacologically counteracting H3K4me3 deficiency repaired OLs and myelin deficits and promoted cognitive recovery in MK801 mice. Taken together, these results demonstrate H3K4me3 status as a new potential target enhancing myelin repair to alleviate SCZ-related cognitive impairment.\u003c/p\u003e","manuscriptTitle":"Perineuronal oligodendrocyte and myelin renewal are epigenetically silenced contributing to cognitive deficits in a murine model of schizophrenia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-04 12:05:52","doi":"10.21203/rs.3.rs-8000305/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"revise","date":"2026-04-21T09:40:40+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"This content is not available.","date":"2026-04-11T00:52:29+00:00","index":2,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2026-03-16T18:29:04+00:00","index":2,"fulltext":"This content is not available."},{"type":"editorInvitedReview","content":"This content is not available.","date":"2026-03-13T17:41:03+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewerAgreed","content":"This content is not available.","date":"2026-03-13T17:13:18+00:00","index":1,"fulltext":"This content is not available."},{"type":"reviewersInvited","content":"","date":"2026-02-02T21:18:20+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-04T11:37:57+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-04T11:17:19+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Psychiatry","date":"2025-10-31T16:25:22+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-psychiatry","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"mp","sideBox":"Learn more about [Molecular Psychiatry](http://www.nature.com/mp/)","snPcode":"41380","submissionUrl":"https://mts-mp.nature.com/cgi-bin/main.plex","title":"Molecular Psychiatry","twitterHandle":"@molpsychiatry","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"00678722-b51a-4793-a20f-2214f534075b","owner":[],"postedDate":"February 4th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[{"id":62192366,"name":"Health sciences/Diseases/Psychiatric disorders/Schizophrenia"},{"id":62192367,"name":"Biological sciences/Neuroscience"}],"tags":[],"updatedAt":"2026-04-21T09:45:45+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-04 12:05:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8000305","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8000305","identity":"rs-8000305","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}