Epilepsy affects over 50 million people worldwide, with more than one-third developing drug-resistant forms. While acupuncture is increasingly utilized for seizure management, its molecular mechanisms remain poorly understood. This study investigated the neuroprotective effects of auricular electroacupuncture (EA) in a kainic acid (KA)-induced rat model of epilepsy. Rats received EA at the heart (CO15) and liver (CO12) acupoints at 2 Hz or 15 Hz for three consecutive days. Although acute seizure behaviors remained consistent across treatment groups-ensuring a homogenous baseline of seizure insult-both EA frequencies significantly reduced KA-induced neuronal apoptosis in the hippocampal CA1, CA2, and CA3 subfields (p < 0.05). Western blot and immunohistochemical analyses showed marker-specific and region-specific changes after EA treatment, including increased hippocampal mGluR5 expression and increased dentate gyrus staining for mGluR5 and GABAA receptor alpha1 subunit. Our findings suggest that auricular EA exerts neuroprotective effects appearing to strengthen the dentate gate and restoring the hippocampal excitatory-inhibitory balance, which might involve mGluR5 and GABAA receptor alpha1 subunit modulation. These results could provide a mechanistic foundation for auricular stimulation as a potential non-pharmacological strategy for drug-resistant epilepsy.

Annexin A1 (ANXA1) is a multifunctional regulatory protein involved in neuroinflammation and cellular homeostasis, but its role in amyotrophic lateral sclerosis (ALS) remains unclear. In this study, we investigated the relationship between ANXA1 and Hippo pathway signaling in ALS. Bioinformatic analysis of transcriptomic data from iPSC-derived motor neurons showed that ANXA1 expression was reduced in ALS and associated with apoptosis and Hippo pathway-related changes. Decreased ANXA1 expression was confirmed in hSOD1^G93A transgenic mice and NSC34 motor neuron-like cells. Phosphorylation levels of MST1/2, LATS1/2, and YAP1 were elevated, indicates activation of the Hippo pathway. Functional experiments showed that ANXA1 overexpression reduced Hippo pathway activation, enhanced cell viability, and decreased apoptosis, as increased Bcl-2 expression and reduced Bax and cleaved caspase-9 levels. In contrast, ANXA1 knockdown further activated Hippo pathway and aggravated apoptotic changes. These findings identify ANXA1 as an upstream regulator associated with Hippo pathway activation in ALS and suggest that the ANXA1/Hippo axis may represent a potential therapeutic target.

Vascular dementia (VaD), the second most common cause of dementia, is closely associated with chronic cerebral hypoperfusion and progressive cognitive decline; however, its molecular and cellular basis remains incompletely understood. Dysregulated intercellular communication within the neurovascular unit may contribute to the cellular and pathological alterations observed in VaD. To identify candidate signaling pathways associated with VaD, we integrated single-cell transcriptomic analysis, microarray data analysis, pseudotemporal trajectory analysis, and experimental validation. Pleiotrophin (PTN) and LAMININ signaling emerged as consistently altered intercellular communication pathways and were associated with microglial state transitions along pseudotime. In a right unilateral common carotid artery occlusion (rUCCAO) mouse model, VaD mice exhibited cognitive impairment, microglial activation, myelin loss, white matter injury, enhanced inflammatory responses, and oxidative stress. Expression analyses further confirmed alterations in PTN- and laminin-related molecules, whereas untargeted metabolomics revealed concurrent disturbances in glutamate-related metabolism. These molecular and metabolic alterations were associated with behavioral deficits and VaD-related pathological changes. Collectively, our findings identify altered PTN/laminin signaling and glutamate metabolic dysregulation as candidate molecular features of VaD and provide a basis for investigating their functional relevance. Pathway-specific functional studies are still required to establish their causal contributions to VaD progression.

Recent work has shown the importance of perineuronal nets (PNNs) in modulating neuronal function and plasticity. Functional studies increasingly manipulate PNNs to probe their role in learning and memory, yet interpreting these manipulations requires knowing which cell types PNNs surround, information that remains incomplete for several brain regions. This study focuses on the prelimbic cortex to determine the percentage of specific interneurons and pyramidal cells that are surrounded by PNNs in male rats and mice. We used immunohistochemistry to assess prelimbic cortex cells containing parvalbumin (PV), glutamate decarboxylase (GAD65/67), and/or calcium kinase II (CamKII), and Wisteria floribunda agglutinin to identify PNNs. Cells with PV, GAD65/67 and/or CamKII were counted and the percentage with PNNs was calculated. Among the cell types examined, PV-expressing interneurons showed the highest frequency of PNN association in the prelimbic cortex of both rats and mice, followed by GAD65/67-expressing interneurons. No CamKII-positive cells had PNNs in rats or mice. PV-only and GAD-only cells were much more likely to have PNNs in mice than in rats. In both species, a small percentage of PNNs surround cells that had no detectable immunolabeling. These findings indicate that PV interneurons are most likely to have PNNs in layers 5/6 of the prelimbic cortex, but species differences suggest caution in designing and interpreting experiments that manipulate PNNs, particularly when generalizing between rat and mouse models.

Stroke disrupts the brain's ability to process and integrate information over time, yet the underlying molecular mechanisms remain unclear. This study investigates post-stroke alterations in intrinsic neural timescales (INTs)-a measure of regional temporal integration-by combining resting-state fMRI, spatial transcriptomics, and PET-based neurochemical mapping. Fifty acute ischemic stroke patients (within 7 days of onset) and fifty matched healthy controls were examined. Voxel-wise and network-level analyses revealed significantly reduced INTs in temporoparietal and insular cortices, with pronounced network-level impairments in the visual and cerebellar systems. Using data from the Allen Human Brain Atlas, we identified gene expression patterns associated with these disruptions. Genes negatively associated with INT reductions were enriched for synaptic, mitochondrial, and neurodevelopmental pathways, while positively associated genes reflected immune signaling and nuclear transport. INT alterations also correlated with excitatory and inhibitory neuronal signatures and were spatially aligned with GABA-A receptor density. Importantly, INT reductions showed significant correlations with clinical assessments: whole-brain INT correlated negatively with NIHSS (r = -0.41) and positively with MoCA (r = 0.38) and FMA (r = 0.35); SMN INT correlated with FMA motor subscore (r = 0.44); and regional INT correlated with domain-specific NIHSS subscores (neglect: r = -0.42; language: r = -0.38). These findings position INT as a clinically meaningful systems-level correlate of stroke-induced dysfunction and highlight molecular pathways and neurotransmitter systems that may constrain temporal integration and recovery potential.

Central nervous system (CNS) disorders, including spinal cord injury, stroke, multiple sclerosis, and motor neuron diseases, are major causes of long-term disability largely due to the persistent failure of axonal regeneration. Over recent decades, diverse strategies have been developed to overcome these barriers, including modulation of extracellular matrix composition, engineering of biomaterial scaffolds, and reactivation of intrinsic growth-associated signaling pathways. However, interventions targeting isolated molecular or environmental factors have produced limited regenerative outcomes. Here, we propose a conceptual framework in which neural repair is understood as the reconstruction of a permissive regenerative niche that aligns intrinsic neuronal growth programs with engineered extracellular microenvironments. We synthesize current knowledge on the cellular and molecular determinants of axonal regeneration and discuss how extracellular matrix architecture and biomaterial scaffolds provide instructive physical and biochemical cues that stabilize growth cones and guide axonal extension. In parallel, intrinsic strategies targeting pathways such as mTOR, STAT3, Krüppel-like factors, and the Lin28/let-7 axis can reestablish neuronal growth competence. Advanced in vitro platforms further enable controlled integration of intrinsic and extrinsic variables, providing experimental models of regenerative niches and testbeds for combinatorial interventions. By framing axonal regeneration as an emergent property of coordinated neuron-environment interactions, this review bridges neurobiology and bioengineering to propose a disease-relevant conceptual framework for neural repair and highlights the need for integrative strategies capable of aligning neuronal state with engineered extracellular landscapes.

Hyperhomocysteinemia (HHcy) is a recognized risk factor for cognitive impairment, including Alzheimer's disease, but the mechanisms linking homocysteine (Hcy) accumulation to cerebral dysfunction remain unclear. Brain microvascular endothelial cells (BMECs) are an important cellular component of the blood-brain barrier (BBB) and play a key role in maintaining central nervous system homeostasis. However, how Hcy affects BMECs remains incompletely understood. In this study, we used the human brain microvascular endothelial cell line hCMEC/D3 to determine the half-maximal inhibitory concentration (IC50) of Hcy by CCK-8 assay, and then performed transcriptomic analysis to identify Hcy-regulated genes associated with endothelial cell injury. Hcy reduced hCMEC/D3 cell viability in a dose-dependent manner, with an IC50 of approximately 12.25 mM; therefore, 10 mM Hcy was selected for subsequent experiments. Transcriptomic sequencing revealed significant upregulation of the transferrin receptor (Tfrc) gene, which we confirmed by quantitative real-time PCR and Western blot analysis. Functionally, TFRC overexpression mimicked the Hcy-induced reduction in cell viability, whereas TFRC knockdown partially rescued cells from Hcy-induced cytotoxicity. TFRC overexpression reduced hCMEC/D3 cell viability, whereas TFRC knockdown partially alleviated Hcy-induced cytotoxicity. These results suggest that TFRC is involved in Hcy-induced injury of brain microvascular endothelial cells. Taken together, our in vitro findings provide a possible clue for future in vivo studies on HHcy-associated BBB dysfunction.

The piriform cortex was identified as a crucial ictogenic trigger zone in the 1980s. Surgical outcome studies in humans likewise suggest that this region contributes to seizure generation. Within the anterior piriform cortex, the "area tempestas (AT)" has been identified as a particularly sensitive site for triggering seizures in rodents and non-human primates. While many studies have examined this region using focal pharmacological manipulations (e.g., drug microinjection) in rodents, within-subject reproducibility has been limited by the need for repeated drug injection into the brain parenchyma. To address this issue, here we developed a chemogenetic model to induce acute seizures in rats by targeting the AT. Twenty-two adult Wistar rats (Rattus norvegicus; 5 males, 17 females; approximately 4 months old, 215-490 g body weight) were used for this purpose. We first examined the efficacy of focal activation of the AT using a glutamatergic neuron targeted excitatory DREADD and compared this to disinhibition using an inhibitory DREADD targeted to GABAergic neurons. The hM4(Gi) construct targeted to GABAergic neurons, but not the Dq construct targeted to glutamatergic neurons induced seizures after clozapine-N-oxide (CNO) administration. CNO (5 or 10 mg/kg) rapidly induced seizures after unilateral or bilateral hM4(Gi) infusion into the AT. Electroencephalographic recordings demonstrated focal seizure onset in the AT which spread ipsilaterally to frontal cortex, then hippocampus, and finally to the contralateral frontal cortex. Synchronized spike bursts were correlated with stage 4 and generalized tonic-clonic seizures. GAD immunofluorescence revealed ≃87% of colocalization with hM4(Gi) receptors, consistent with seizures induced via reduction of inhibitory input in the AT. These results suggest that the present model provides controlled, localized, and reproducible seizure induction with low (‹10%) mortality rates.

Spinal cord injury (SCI) leads to persistent motor deficits, and effective strategies to enhance neural repair remain limited. Repetitive transcranial magnetic stimulation (rTMS) has been reported to promote neuroplasticity, but its effects on corticospinal remodeling after SCI are not fully understood. In this study, adult female rats were assigned to Sham, SCI, or rTMS groups. High-frequency rTMS was applied daily for 28 days starting 1 day after injury. Motor function was assessed using Basso-Beattie-Bresnahan scoring and gait analysis. Corticospinal conduction was evaluated by motor evoked potentials (MEPs). Histological changes, plasticity-related proteins, and MEK/ERK signaling were examined in the spinal cord and primary motor cortex (M1). Corticospinal tract (CST) remodeling was evaluated using anterograde tracing. rTMS significantly improved locomotor recovery and gait coordination, enhanced MEP responses, and reduced tissue damage compared with SCI controls. These functional improvements were accompanied by increased CST projections distal to the lesion. rTMS also increased phosphorylation of MEK and ERK, together with partial restoration of GAP-43, SYN1, and PSD-95 expression in both spinal cord and M1. These findings indicate that high-frequency rTMS facilitates functional recovery after SCI, which is accompanied by enhanced corticospinal remodeling and plasticity-related signaling.