Alzheimer’s disease (AD), the most common form of dementia, is characterized by cholinergic dysfunction and early impairments in episodic and recognition memory. Although retrieval failure represents a hallmark of cognitive decline in AD, the task- and phase-specific molecular mechanisms underlying cholinergic-dependent memory retrieval remain poorly defined. This study investigated whether acute muscarinic cholinergic blockade disrupts the retrieval phase of recognition memory. We also examined the associated regulation of hippocampal calcium/calmodulin-dependent protein kinase II (CaMKII) isoforms α and β, together with extracellular signal-regulated kinase (ERK) signaling. Adult male SWR/J mice were trained in the NOR paradigm and administered scopolamine (1 mg/kg, i.p.) or saline 30 min prior to the retrieval phase. Recognition memory performance was assessed using the discrimination index and exploratory preference. Immediately following behavioral testing, hippocampal tissue was collected for Western blot analysis of phosphorylated and total CaMKII-α, CaMKII-β, and ERK. Scopolamine significantly impaired recognition memory retrieval, as evidenced by a reduced discrimination index and decreased preference for the novel object, without diminishing overall exploratory activity. At the molecular level, scopolamine-induced cholinergic inhibition selectively decreased hippocampal phosphorylation of both CaMKII-α and CaMKII-β while increasing ERK phosphorylation, with no significant changes in total protein expression of any kinase. These findings identify a retrieval-specific, scopolamine-induced molecular signature of cholinergic inhibition in the hippocampus, characterized by concurrent suppression of both CaMKII isoform activities (α and β) alongside dissociable ERK hyperactivation. By extending prior molecular observations from aversively motivated paradigms to an ethologically relevant model of recognition memory, this study provides mechanistic insight into how cholinergic dysfunction contributes to memory retrieval deficits and suggests that cholinergic signaling maintains the functional (CaMKII-α) and structural (CaMKII-β) dimensions of hippocampal memory retrieval machinery through coordinated kinase activation.
Aberrant protein phosphorylation and sustained neuroinflammatory activation are key biochemical features of experimental parkinsonism. Dysregulated tau processing and excessive microglial reactivity have been implicated in motor dysfunction associated with dopaminergic injury. In the present study, we evaluated the biochemical and functional effects of Leucettine L41 using biochemical, cellular, and in vivo systems. In an in vitro tau aggregation assay, L41 reduced the ThT fluorescence, indicating the suppression of fibril formation. In SH-SY5Y neuronal cells, L41 decreased tau phosphorylation at the Ser202/Thr205 site. In lipopolysaccharide-stimulated BV-2 microglial cells, L41 suppressed the production of nitric oxide, TNF-α, IL-1β, and IL-6, demonstrating its anti-inflammatory activity. Notably, L41 did not prevent toxin-induced neuronal cytotoxicity, suggesting that its primary action may not involve direct neuroprotection. In an MPTP-induced mouse model of Parkinson’s disease, systemic L41 administration improved motor performance, accompanied by reduced tau phosphorylation and attenuated microglial activation in brain tissue. These findings indicate that L41 modulates tau aggregation, phosphorylation, and neuroinflammatory signaling in experimental parkinsonism, in association with functional motor improvement.
Intracerebral hemorrhage (ICH) is a health challenge resulting in death or disability. Iron overload has been identified as one of post-ICH damaging factors. However, the impact of iron overload, as well as the underlying mechanisms, on neural stem/progenitor cells (NSPCs) remains unknown. In this study, we found that iron overload induced significant NSPC death in a dose-dependent manner. Interestingly, iron overload increased the number of early and late apoptotic cells and the expression of caspase-3 in NSPCs. Moreover, iron overload was highly correlated with enhanced intracellular reactive oxygen species (ROS) generation and loss of mitochondrial integrity. Further, iron overload activated mTOR signaling and downregulated AMP-activated protein kinase (AMPK) phosphorylation. Importantly, drugs that eliminate ROS or activate AMPK prevented this event, as well as apoptosis of NSPCs. These results indicate that post-ICH iron overload in the brain induces excessive ROS generation and leads to NSPC death by regulating mTOR and AMPK signaling.
Extensive research with rodent models has shown detrimental effects of early-life adversity (ELA) on behavioral (e.g., impulsive behavior, anxiety, and depression) and neurobiological processes (e.g., alterations of neuroendocrine processes and maturation of brain areas). However, heterogeneous methodologies, including types and variations of ELA manipulations could have contributed to inconsistent findings across studies. Recent research indicates that the combination of the two most widely implemented rodent ELA protocols, Maternal Separation (MS) and Limited Bedding/Nesting (LBN), produces consistent and robust behavioral effects. We assessed the effects of combined MS-LBN on four processes linked to both ELA and behavioral disorders in later stages of life: incentive salience of reward cues, and impulsive choice, action, and persistence/perseverance. Sixteen male Sprague Dawley rats were divided in groups of combined MS-LBN during postnatal days 2–21 and without ELA. They were exposed to an Autoshaping Pavlovian conditioning task (AUT), a delay-discounting task (DDT), and acquisition and extinction of a multiple schedule of reinforcement with long and short Variable Intervals (VI) across a span of 18 weeks. Compared to the No-ELA group, ELA rats displayed higher goal-tracking during the AUT (higher nose-poking in the food-delivery location), higher impulsive choice during test and 7-week re-test of the DDT (preference for the smaller-sooner reinforcer over the larger-later), and less efficient responding during the long-interval schedule of reinforcement (more unnecessary responses per pellet). Associations between autoshaping and reinforcement-schedule performance were identified, with ELA moderating the relationship between sign tracking and efficiency during training and persistence/perseverance during extinction. Combined MS-LBN was associated with task-specific changes in reward-related and impulsive behavior in male rats. These complex patterns of disruptions across behavioral processes adds further support to the notion that combined ELA protocols are promising robust models of adverse rearing in humans, which often entails multiple stressors. Not applicable.
Cisplatin-induced neurotoxicity is driven in part by neuroinflammation and oxidative injury in vulnerable brain regions. Glycine has anti-inflammatory and antioxidant properties that may offer neuroprotection against chemotherapy-related brain damage. Twenty-five adult male BALB/c mice were randomized into five groups (n = 5/group) Group 1 received cisplatin for 14 days; Group 2 received cisplatin plus glycine for 14 days; Group 3 received cisplatin for 28 days; Group 4 received cisplatin for 14 days followed by glycine for 14 days; and Group 5 received cisplatin for 28 days with glycine introduced from day 14 to day 28. Cisplatin was administered intraperitoneally at 3 mg/kg every fourth day, and glycine was given subcutaneously at 1 g/kg daily. The primary outcome was serum TNF-α measured by ELISA. Secondary outcomes were neuronal integrity and optical density in the hippocampus and frontal cortex assessed by Nissl staining. Data were analyzed using one-way ANOVA with Tukey post-hoc testing. Serum TNF-α levels differed significantly among groups (F = 230.422, p < 0.001). Mean TNF-α concentrations were 150.0 pg/mL in Group 1, 130.2 pg/mL in Group 2, 201.4 pg/mL in Group 3, 159.4 pg/mL in Group 4, and 171.0 pg/mL in Group 5. Prolonged cisplatin exposure (Group 3) produced the highest TNF-α levels, whereas concurrent glycine administration during the 14-day regimen (Group 2) resulted in the lowest levels. Compared with the 28-day cisplatin group, both delayed glycine treatment (Group 4) and glycine introduced during the second half of cisplatin exposure (Group 5) were associated with lower TNF-α concentrations. Histological analysis demonstrated reduced Nissl staining intensity and neuronal preservation in cisplatin-only groups, particularly Group 3, whereas glycine-treated groups showed better preservation of neuronal architecture and optical density in the hippocampus and frontal cortex. Glycine attenuated cisplatin-induced neuroinflammation and preserved neuronal integrity in the hippocampus and frontal cortex of mice. These findings support further preclinical evaluation of glycine as a low-cost adjuvant strategy to reduce chemotherapy-associated neurotoxicity.
Abstract Due to delayed symptoms and dependence of behavioral assessment, early diagnosis of autism spectrum disorder remains challenging. Identification of multivariate biomarker for the etiological mechanisms of ASD may enhance diagnostic accuracy. Multivariable logistic regression combines many predictors into a single risk score (linear predictor), resulting in an optimised ROC curve that enhances diagnostic accuracy over individual markers. The method comprises modelling a binary result, determining the likelihood, and visualising ROC based on the projected probabilities, which often improves individual marker AUCs. In the present study a diagnostic performance for a biomarker panel reflecting glutamatergic dysfunction, oxidative stress, and neuroinflammation was evaluated. Plasma levels of glutaminase, 8-isoprostane, and prostaglandin E₂ (PGE₂) obtained from 44 children with ASD and 40 age-matched controls were evaluated using receiver operating characteristic (ROC) analysis, both individually and in combined ROC models. Glutaminase showed significant negative correlations with both 8-isoprostane and PGE₂, whereas a positive correlation was observed between 8-isoprostane and PGE₂. All the three-biomarker showed good diagnostic performance for ASD on its own with statistically significant ( p = 0.001) values of AUC of 0.830 for glutaminase, AUC of 0.815 for 8-Isoprostane and AUC of 0.818 for PGE₂. However combined ROC modeling substantially improved diagnostic accuracy by achieving high apparent discriminative performance with AUC value of 0.977 with 92.3% sensitivity and 100.0% specificity. In conclusion, the diagnostic usefulness of independent glutaminase, 8-isoprostane, and prostaglandin E₂ (PGE₂) biomarkers may be enhanced by combining ROC. Combined markers show strong apparent discriminating power in a case-control method, but estimates are biassed towards optimism and are not diagnostic. Comprehensive assay validation, calibration, and clinically representative cohorts (including females and relevant differentials) are required for replication.
The CHRNA7 gene, located on chromosome 15q13.3, encodes the α7 nicotinic acetylcholine receptor (α7 nAChR) subunit and lies within a genomic region characterized by high recombination rates and associations with multiple neuropsychiatric disorders. Within this region, the human specific gene CHRFAM7A arose through partial duplication, rearrangement, and fusion between CHRNA7 and FAM7A. The direct CHRFAM7A allele has been shown to negatively regulate α7 nAChR function. The inverted allele (CHRFAM7AΔ2bp), harboring a two-base pair deletion in exon 6, is linked to schizophrenia, bipolar disorder, and other psychiatric conditions. In this study, we investigated how the presence of the CHRFAM7AΔ2bp allele alters the cellular proteome. Using high-throughput proteomic analysis by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), we characterized protein expression in neuronal progenitors differentiated from isogenic human induced pluripotent stem cell (iPSC) lines representing CHRFAM7AΔ2bp and CHRFAM7A-null genotypes. Comparative analysis identified 129 differentially expressed proteins enriched in pathways related to extracellular matrix organization, collagen biosynthesis, and cell adhesion. Functional assays further demonstrated differences in matrix adhesion between CHRFAM7A-null and CHRFAM7AΔ2bp-derived progenitors. These results suggest that the CHRFAM7AΔ2bp variant influences cellular structure through modulation of adhesion matrix-interaction proteins. This work provides insight into molecular mechanisms that may underlie increased neurodisease vulnerability associated with this genotype.
Abstract Background Despite neuroinflammation being an initially protective response made by the central nervous system (CNS), as it becomes chronic, it can lead to neuronal damage since the cytokines which are released by microglia potentialize cellular death due to excitotoxicity, and this, in turn, promotes the release of pro-inflammatory mediators, feeding this way a self-sustaining cycle of neuroinflammatory response, which favor subsequent neurodegeneration. Given the fact that until this moment, there are not any therapeutic alternatives able to stop the neurodegeneration, the objective of the present work was to evaluate the putative neuroprotector effect of taurine, a partial glycinergic ionotropic receptor agonist, and also a GABA A receptor agonist, in a neuroinflammation animal model. Method For this intent, the oral taurine administration was evaluated on mnemonic impairing caused by LPS induced neuroinflammation in male Wistar rats. Such effects were investigated on recent and late spatial long-term memory and aversive memory in the behavioural tasks Morris water maze (MWM) and context fear conditioning (CFC), respectively. In addition, we investigated the effect of orally administered taurine on hippocampal neuronal density and on hippocampal levels of TNF-α and IL-4. Results Taurine, when orally administered for 30 days, in the doses of 20 and 200 mg/kg, was able to reverse the mnemonic impairment caused by neuroinflammation on recent and remote spatial long-term memory, and in the dose of 200 mg/kg, it also was able to do the same on aversive long-term memory. In the doses of 20 and 200 mg/kg, taurine was also able to reverse the LPS-induced increase in hippocampal TNF-α levels. Conclusion Taurine, when orally administered in a pathological context characterized by neuroinflammatory background, as it was induced in this work, can perform a dose-dependent neuroprotective effect, probably by acting in an excitotoxic scenario in which the activation of hyperpolarizing receptors can be welcomed.
Cerebral toxoplasmosis is a common opportunistic parasitic infection of the CNS caused by the Toxoplasma gondii parasite. Host immunosuppression can affect disease outcomes. To explore the changes in the cerebral cortical ultrastructure accompanying the infection in different immune-altered models and to find an effective treatment against the infection, we tested the possible therapeutic effect of clofazimine (CFZ) (the FDA-approved antimycobacterial drug) against the infection using 60 male CD1 Swiss Albino mice divided into 6 groups: 3 dexamethasone (DEX)- treated groups (DEX-only, DEX-infected, and DEX-infected-treated), and 3 streptozotocin (STZ)-induced type 1 diabetic groups (STZ-only, STZ-infected, STZ-infected-treated). The worst ultrastructural changes were observed in the diabetic and diabetic-infected groups, characterized by a significant increase in neuronal apoptotic and necrotic nuclei (P < 0.05) and changes in the numbers and structure of glial cells compared to the DEX and DEX-infected groups. CFZ (at a dose of 10 mg/kg/day for 3 days starting on 45th day post infection) significantly improved cortical neuronal ultrastructural changes in both models (P < 0.05), reduced microglial numbers, increased astrocyte numbers, and restored brain capillary integrity and axonal growth, in addition to significantly reducing mature cyst numbers in both models (P < 0.05). However, the drug didn’t reduce the number of atrophic and necrotic cysts in the infected-treated groups. So, in our study, CFZ showed preclinical promise in treating experimental cerebral toxoplasmosis and reducing the parasitic cyst burden, highlighting the adverse impact of the host’s altered immune status on brain tissue and the course of the infection, especially in diabetes.
The intersection of age- and experience-dependent processes influence learning throughout development, and developmental learning can have long-term effects on behavior. Phosphorylation of ribosomal protein S6 phosphorylation (pS6) is required in active ribosomes, and new protein synthesis is a conserved mechanism that supports long term memory formation. As such, pS6 fluctuations in brain regions processing experience can provide insight into shifts in the ability for learning and memory. Juvenile male and female zebra finch songbirds (Taeniopygia guttata) perform sensory song learning in ways that affect their adult behaviors. As adults, both sexes perform song recognition learning. Both juvenile and adult types of sensory learning invoke the auditory forebrain. Prior reports established a pS6 song response in the auditory forebrain in Posthatch day 30 juvenile males but not females, and not in younger birds. This was intriguing because the experience-dependent pS6 increase tracked with the onset of the critical period for juvenile sensory song learning in males, and behavioral data indicated that females also effectively learn at P30, though they may not have a critical period. Further, by adulthood (> Posthatch day 90), both male and female auditory forebrains showed an equivalent increase in pS6 after hearing song in patterns consistent with effective recognition learning. Here, to further test the relationships between a crucial molecular marker of active learning processes and the developmental trajectory of juvenile sensory song learning and the emergence of adult-like song recognition learning, we assessed the effect of age and sex, as well as the absence of tutor experience, a manipulation that extends the critical period for developmental learning in males, on the phosphorylation of S6 within the auditory forebrain. Outcomes highlight the complexity of molecular mechanisms across developmental learning and reveal questions to be addressed by further inquiry.
High-intensity interval training (HIIT) has been proposed as a time-efficient strategy to enhance brain health, yet its longer-term neural signatures remain unclear. In a randomized, assessor-blinded, parallel-group study, 32 healthy undergraduates (18–25 years) completed either 8 weeks of supervised HIIT (30 min/session, 3 sessions/week) or a usual-activity control. Body composition was measured before and after the intervention, and maximal oxygen uptake (VO2max) was assessed using the 20-m shuttle run test (20 m SRT). Resting-state EEG was recorded at baseline and post-intervention. Power spectral density (PSD) and phase-based connectivity were quantified across canonical frequency bands. HIIT increased VO2max and skeletal muscle mass and reduced body fat percentage compared with baseline, while no meaningful changes were observed in controls. Resting-state EEG showed increased delta-theta-alpha power and inter-regional phase synchronization, alongside reduced beta-gamma power/connectivity. 8 weeks of HIIT were associated with frequency-specific modulation of cortical oscillations and large-scale functional coupling at rest. These findings suggest that time-efficient exercise paradigms may promote neural network reorganization, with potential benefits for cognitive and motor function.
Overexpression of Sxc-, a cystine/glutamate antiporter is implicated in several neurological disorders including epilepsy. Physiologically, it regulates the hippocampal synaptic glutamate pool, influencing the neuronal excitability. Sulfasalazine (SAS) recently garnered interest owing to its system xc- (Sxc-) inhibitory potential. Therefore, the current study investigated the effects of SAS in two distinct models, (i) maximal electroshock (MES) induced acute seizure model and (ii) Pentylenetetrazol (PTZ) induced kindling model of chronic epilepsy and associated cognitive impairment. Initially, rats treated with different doses of SAS (25- 200 mg/kg, i.p.) for 7 days were subjected to MES and observed for seizures. SAS 50 mg/kg improved the extension-to-flexion ratio (E/F ratio) in MES induced seizures and exhibited neuroprotection, whereas the higher doses failed. In another experiment, mice were treated with PTZ (35 mg/kg) on alternate days for 24 days to induce kindling. SAS (50 or 100 mg/kg, i.p.) or vehicle was administered daily, 30 min prior to each PTZ injection. Seizures were monitored for 30 min following PTZ injection. Chronic PTZ administration led to progressively increased seizure severity, culminating in generalized tonic-clonic seizures in mice. SAS 100 mg/kg treatment reduced stage 4 and 5 seizure scores and improved spatial retention memory in mice. SAS also significantly attenuated the gene expression of inflammatory cytokines, TNF-α, IL-1β, and IL-6, and mitigated neuronal damage induced by epileptic kindling. Altogether, results suggest that SAS exhibits anti-epileptic effects, reduces neuroinflammation, and alleviates cognitive dysfunction in the PTZ kindling model of epilepsy in mice.
Synesthesia refers to a neurological condition in which stimuli from one sensory pathway induce perception via another sensory pathway. This study evaluated self-reported synesthesia related experiences, awareness of such experiences, and other associated demographic and clinical attributes among undergraduate medical students. This cross-sectional online survey targeted 617 medical students aged 18–25 years during the 2024–2025 academic year. Data were collected using a structured questionnaire on demographics, awareness of synesthesia-related experiences, selected medical and psychological factors, and their possible relevance to education. All analyses were performed using IBM SPSS Statistics (version 25.0). Among the 617 participants, 53.5
Leiotrametes lactinea (Berk.) Welti Courtec. a mushroom reported to possess antimicrobial, antioxidant, antitumor and analgesic activities. However, there is limited information on its safety profile in the central nervous system. This study thus evaluates the neurotoxic effect of ethanol extract of Leiotrametes lactinea (EELL) in male Swiss mice following 30 days oral exposure. Twenty-four (24) Swiss male mice were divided into 4 groups (n = 6) and orally administered EELL for 30 days at doses of 50, 100, and 200 mg/kg; the last group served as the healthy control. Neurobehavioral assessment started on the 21st day, and on day 30, the animals were euthanised. Brain tissues were collected for biochemical assays (Superoxide dismutase, Catalase activity, reduced glutathione, malondialdehyde and acetylcholinesterase activity). Tumour necrosis factor-alpha and interleukin-6 levels were also measured. There was a reduction in body weight gain of EELL administered groups (3.38 ± 0.68 - 4.28 ± 0.85 g) compared to the control group (7.21 ± 0.87 g), while relative organ weights were unaffected. Also, a significant reduction in locomotor activity and rearing behaviour was recorded with an increased anxiety-like behaviour in the extract treated groups. Cognitive performance evaluated with the Morris water maze demonstrated impaired retention, evidenced by reduced time spent in the target platform zone and increased proximity to the platform at higher doses. Biochemical evaluation of brain tissue showed pronounced oxidative stress, indicated by significant reductions in reduced glutathione (71.06 ± 4.74 - 124.36 ± 5.35 µg/mol vs 149.88 ± 7.07 µg/mol), catalase, and superoxide dismutase activities, alongside elevated malondialdehyde levels, especially at 100 and 200 mg/kg doses in comparison to the control. In addition, acetylcholinesterase activity was significantly increased in 100 and 200 mg/kg EELL treated groups compared to the control (0.121 ± 0.01 0.160 ± 0.01 µmol/min/g tissue vs. 0.049 ± 0.01 µmol/min/g tissue). Likewise, increased levels of pro-inflammatory cytokines (interleukin-6 and tumour necrosis factor-α) was recorded in the treated groups. These findings demonstrated that sub-acute exposure to EELL induces neurobehavioral deficits, oxidative stress, and neuroinflammatory responses, suggesting potential neurotoxic effects at higher doses.
Neuroinflammation is a hallmark of numerous neuropsychiatric and neurodegenerative disorders. Psychedelics have garnered recent attention for their anti-inflammatory and neuroprotective effects. However, it remains unknown whether they can prophylactically prime immune pathways to prevent subsequent neuroinflammatory responses, and whether such effects depend on serotonin 5-HT2A receptor (5-HT2AR) signaling. To characterize the inflammatory time-course, wild-type (WT) male mice received LPS (1 mg/kg, i.p.) and hippocampi were collected 2, 4, 6, 8, or 24 h later. In a separate WT cohort, basal cytokine effects of the psychedelic (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) were examined following administration of varying doses or vehicle, with tissue collected 24 h later. For prophylactic studies, WT and 5-HT2AR-KO mice were pretreated with DOI (0.3 or 1 mg/kg) or saline 24 h before LPS challenge. Behavioral assays included locomotor activity, the forced swim test (FST), and body weight monitoring. Hippocampal cytokines (IL-6, TNF-α, IFN-γ, IL-5, IL-4, IL-10, IL-13, IL-2) were quantified with a multiplex bead-based assay, and correlations between cytokine levels and FST immobility were assessed. LPS induced robust hippocampal increases in IL-6 and TNF-α and produced sickness-related behavioral deficits. DOI pretreatment (0.3 mg/kg, and to a lesser extent 1 m/kg) attenuated LPS-evoked IL-6 and TNF-α elevations, restored locomotor activity, reduced FST immobility, and accelerated recovery of body weight. These anti-inflammatory effects were partially preserved in 5-HT2AR-KO mice, indicating both receptor-dependent and receptor-independent mechanisms. Notably, 5-HT2AR-KO mice displayed exaggerated cytokine responses to LPS relative to WT animals. Correlation analyses revealed a positive association between hippocampal IL-6 levels and depressive-like behavior, whereas IL-13 and IL-2 levels were inversely correlated with immobility time in the FST. Prophylactic DOI administration establishes a sustained neuroimmune state that mitigates subsequent hippocampal inflammation and behavioral impairments, through mechanisms that are partially dependent on 5-HT2ARs. These findings suggest that serotonergic psychedelics can prime neural-immune interactions to confer long-lasting resilience against neuroinflammatory insults, offering a potential framework for developing next-generation psychedelic therapeutics with prophylactic anti-inflammatory and neuroprotective efficacy in neuropsychiatric and neurodegenerative disorders.
Alzheimer’s disease (AD) is characterized by progressive neurodegeneration driven by amyloid-β (Aβ)–associated oxidative stress, mitochondrial dysfunction, and dysregulated inflammatory signaling. Although microRNAs (miRNAs) represent promising regulators of these interconnected pathways, their therapeutic application is limited by instability and inefficient cellular delivery. In this study, the cytoprotective potential of milk-derived small extracellular vesicles (sEVs) loaded with miR-126-3p was evaluated in an Aβ-induced SH-SY5Y neuroblastoma cell model. sEVs were isolated and characterized according to MISEV guidelines, loaded with synthetic miR-126-3p, and administered to Aβ-induced cells. miR-126-3p–enriched sEVs significantly attenuated Aβ-induced oxidative stress, as evidenced by normalization of ROS, LDH, GPX1, MDA, and SOD levels, while naïve sEVs exerted only partial effects. At the transcriptional level, miR-126-3p delivery restored stress-responsive gene expression patterns by reducing ICAM1 and TNF-α expression and normalizing BDNF levels, reflecting modulation of neuron-intrinsic inflammatory signaling rather than tissue-level neuroinflammation. Markers of cytoskeletal and mitochondrial stress, including intracellular NfL, cytochrome c, 8-OHdG, TFAM, PINK1, and DNM1L, were also significantly reduced following miR-126-3p–loaded sEV treatment, indicating improved cellular homeostasis under amyloid stress. Furthermore, intracellular tau-related markers and Aβ1–40 accumulation were attenuated, consistent with suppression of Aβ-triggered pathological signaling cascades. Collectively, these findings demonstrate that sEV-mediated miR-126-3p delivery confers robust cytoprotective effects at the cellular level in a neuroblastoma model. While extrapolation to in vivo neurodegeneration requires caution, the results highlight milk-derived sEVs as a biocompatible and scalable platform for miRNA-based modulation of AD-relevant cellular stress pathways.
Spinal cord injury (SCI) is a devastating condition with high rates of disability and mortality, and secondary injury mechanisms such as ferroptosis have emerged as crucial contributors to its pathophysiology. However, the regulatory mechanisms underlying ferroptosis in SCI remain largely unclear. We analyzed peripheral blood transcriptome data (GSE151371) from 10 healthy controls, 10 non-CNS trauma patients, and 38 SCI patients to identify differentially expressed mRNAs (DE-mRNAs) and miRNAs (DE-miRNAs). Ferroptosis-related genes (FRGs) were screened by intersecting DE-mRNAs with a curated list of FRGs. Functional enrichment analyses, regulatory network construction (lncRNA–miRNA–mRNA, TF-miRNA-mRNA), and drug target predictions were conducted. Key findings were validated using the GSE166009 dataset from a rat SCI model and through experimental verification in a TSCI rat model. We identified 38 differentially expressed FRGs (DE-FRGs) in SCI, enriched in lipid metabolism and inflammatory pathways, including IL-17 and TNF signaling. Protein–protein interaction network analysis identified IL-1β as a central hub gene. A miRNA–mRNA network revealed miR-326 as a key regulator of IL-1β. The IL-1β/miR-326 axis was also closely associated with lncRNA-miRNA-mRNA networks, transcription factor regulation, and drug target predictions. Validation in the GSE166009 dataset and a rat SCI model confirmed progressive downregulation of miR-326 and upregulation of IL-1β post-SCI. Dual-luciferase reporter assays demonstrated that miR-326 directly binds the 3′UTR of IL-1β, suppressing its expression. Our study highlights the IL-1β/miR-326 axis as a critical regulator of ferroptosis in SCI and a potential therapeutic target for intervention.
Methamphetamine (METH) abuse is associated with profound cognitive and behavioral impairments, notably in decision-making and social interaction. Emerging evidence suggests the endocannabinoid system, particularly CB1 receptors (CB1Rs), plays a modulatory role in these processes. This study investigated whether pharmacological modulation of CB1Rs can alter METH-induced deficits in decision-making, social behavior, and anxiety in a rodent model. Adult male Wistar rats were administered METH and treated with either a CB1R agonist WIN55,212-2 (WIN; 3 mg/kg; i.p) or antagonist (Rimonabant;1 mg/kg; i.p.). Behavioral assessments included effort-based decision-making via the Y-maze barrier task, social behavior tests, and anxiety evaluation using the elevated plus maze. METH exposure significantly reduced high-reward choices in effort-based tasks and impaired multiple domains of social behavior. It also induced anxiety-like behaviors. Rimonabant treatment partially reversed these effects, improving decision-making performance, enhancing social interactions, and reducing anxiety-related responses. In contrast, WIN did not significantly alleviate METH-induced behavioral impairments. CB1R antagonism via Rimonabant may mitigate METH-induced cognitive and social deficits, underscoring the therapeutic potential of targeting the endocannabinoid system in stimulant-related neuropsychiatric disorders. These findings highlight CB1R antagonists as promising agents for future interventions in METH addiction and its associated behavioral disruptions. Not applicable.
Background The glycinergic system constitutes a main source of inhibitory regulation in the central nervous system. Glycine transporters (GLYT1 and GLYT2), encoded by Slc6a9 and Slc6a5, respectively, are responsible for glycine reuptake and clearance from the synaptic cleft, thereby maintaining neurotransmitter homeostasis. Emerging evidence from pharmacological and mechanistic studies has highlighted GLYTs as promising therapeutic targets for psychiatric disorders and persistent pain. Nevertheless, data on anatomical and cellular distribution of GLYTs and sex-dependent differences in GLYT expression remain limited. Methods To address this gap, the aim of this study was to examine the Slc6a9 and Slc6a5 mRNA expression across mouse brain regions and peripheral organs using three complementary approaches focusing on mRNA expression: re-analysis of single-cell RNA sequencing data, quantitative RT-PCR, and RNAscope. Results Both genes were detected in multiple brain regions, with Slc6a9 exhibiting a broader distribution in both glial cells and neurons, while Slc6a5 was more restricted to neurons. Sex-dependent differences were detected for Slc6a9 in the amygdala and thalamus, liver, intestine, spleen, kidney and genitalia using quantitative RT-PCR, and for Slc6a5 in the cortex, striatum, hippocampus, and spinal cord using quantitative RT-PCR. Spatial analysis of the glycine transporters showed that Slc6a9 can be found in several brain regions spanning the rostral to the caudal axis, in both glial cells and neurons, while Slc6a5 was more restricted to the caudal brain regions. Conclusions In general, in regions where differences were detected using quantitative RT-PCR, higher expression levels were observed in male mice. Moreover, Slc6a9 expression was found to occur in both glial cells, such as astrocytes, oligodendrocytes and ependymal cells, as well as both excitatory and inhibitory neurons, while Slc6a5 mainly occurred in inhibitory neurons. These findings provide novel insights into the spatial and sex-dependent expression of glycine transporters.
Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by microtubule destabilization, neuroinflammation, and tau pathology. Among the proposed therapeutic approaches, acetyl-11-keto-β-boswellic acid (AKBA), a bioactive triterpene from Boswellia serrata, has gained attention due to its multiple neuroprotective mechanisms, including microtubule stabilization, anti-inflammatory activity, antioxidant effects, and promotion of neurogenesis. In this study, we aimed to investigate the neuroprotective effect of AKBA against tau oligomer-induced cytotoxicity in SH-SY5Y neuroblastoma cells. Recombinant human tau protein was expressed, purified, and oligomerized, and the formation of oligomers was confirmed by thioflavin T fluorescence and dynamic light scattering (DLS). SH-SY5Y cells were then treated with AKBA and exposed to tau oligomers. Cell viability was assessed via MTT assay, and apoptosis was evaluated by flow cytometry. The morphology of tau aggregates was visualized using transmission electron microscopy. Our findings demonstrated that AKBA significantly reduced tau oligomer-induced cytotoxicity and enhanced cell viability. These results suggest that AKBA, through its multifaceted protective mechanisms, holds promise as a potential therapeutic agent for the treatment of tauopathies such as Alzheimer’s disease. Not applicable.
