Journal of Neuroscience Methods最新文献

{"title":"Multiscale spatiotemporal neural network with multi-attention mechanism using brain partitioning for motor imagery recognition","authors":"Moeed Sehnan ,&nbsp;Haoyu Li ,&nbsp;Xiaoyang Li ,&nbsp;Celso Grebogi ,&nbsp;Zhongke Gao ,&nbsp;Weidong Dang","doi":"10.1016/j.jneumeth.2026.110704","DOIUrl":"10.1016/j.jneumeth.2026.110704","url":null,"abstract":"<div><h3>Background:</h3><div>Motor imagery (MI)-based electroencephalogram (EEG) brain-computer interfaces (BCIs) facilitate communication for motor-impaired patients by leveraging artificial intelligence to accurately interpret brain signals. However, EEG signal classification remains challenging due to low signal-to-noise ratio (SNR) and individual variability in brain activity.</div></div><div><h3>New method:</h3><div>We propose a novel parallel multi-depth spatial–temporal neural network aimed at enhancing the integration of spatial and temporal features from multichannel EEG signals by leveraging brain functional topography. To improve cortical representations associated with motor imagery, the model incorporates two parallel branches. One branch focuses on inter-channel differences corresponding to contralateral electrode pairs, emphasizing hemispheric disparities, while the other targets the frontal and parietal brain regions. These region-specific enhanced signal representations are then fed into the multi-depth spatial–temporal network for feature extraction and subsequent motor imagery classification. The architecture of the feature extraction network integrates four specialized blocks, ensuring the comprehensive capture of discriminative features that are particularly sensitive to task-relevant frequencies for each MI class. A multi-loss design further optimizes feature integration across networks.</div></div><div><h3>Results:</h3><div>Cross-validation results on the BCI Competition IV 2a dataset and High Gamma dataset achieve accuracies of 82.14% and 95.61%, respectively, with kappa values of 0.76 and 0.93, surpassing state-of-the-art methods.</div></div><div><h3>Conclusion:</h3><div>These experimental results highlight the significance of parallel spatial–temporal networks based on brain partitioning for MI classification in rehabilitation engineering and real-world BCI applications.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110704"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146125325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiple epileptiform waves detection algorithm based on improved VMD and multidimensional feature fusion","authors":"Qiwei Cai ,&nbsp;Dinghan Hu ,&nbsp;Feng Gao ,&nbsp;Xiaohui Lou ,&nbsp;Jiuwen Cao","doi":"10.1016/j.jneumeth.2026.110703","DOIUrl":"10.1016/j.jneumeth.2026.110703","url":null,"abstract":"<div><h3>Background:</h3><div>Spikes, ripples, and ripples on spikes (RonS) during non-rapid eye movement (NREM) sleep are all important biomarkers associated with epileptic seizures, and accurate detection of these epileptiform waves is vital for epilepsy analysis.</div></div><div><h3>New Method:</h3><div>An improved variational mode decomposition (VMD) decomposes frequency bands to isolate target epileptiform waves. Multidimensional handcrafted features are extracted from low and high frequency bands to characterize these waves, with recursive feature elimination (RFE) selecting key ones. Meanwhile, a dual-stream 1-dimension convolutional neural network (1D CNN) with an adaptive scale factor extracts deep features from VMD-decomposed bands, which are then fused with the handcrafted features.</div></div><div><h3>Results:</h3><div>Experimental results show that the proposed algorithm achieves an average precision of 91%, a recall of 90.36%, and an F1-score of 90.62% on the scalp electroencephalogram (EEG) data of 16 children with benign childhood epilepsy with centrotemporal spikes (BECTS) from the Children’s Hospital of Zhejiang University School of Medicine (CHZU).</div></div><div><h3>Comparison with existing methods:</h3><div>Previous studies have often focused on only one type of epileptiform discharge. This narrow focus limits the translation of these biomarkers into clinical practice and their comprehensive application. In the present study, three types of epileptiform discharges are focused on simultaneously.</div></div><div><h3>Conclusion:</h3><div>Our method achieves the optimal overall detection performance in the detection of multiple epileptiform waves. It can be concluded that the proposed technique is capable of serving as an effective tool for evaluating multiple epileptiform waves.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110703"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146093141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neuroimaging of reality: A new approach for investigating neural bases of decision-making with real-world objects","authors":"Damien Gabriel ,&nbsp;Guillaume Bertrand ,&nbsp;Magali Nicolier ,&nbsp;Julie Giustiniani","doi":"10.1016/j.jneumeth.2026.110692","DOIUrl":"10.1016/j.jneumeth.2026.110692","url":null,"abstract":"<div><h3>Background</h3><div>Neuroimaging studies often use computerized tasks, but reliance on virtual stimuli limits ecological validity. Incorporating real object interaction under controlled recording conditions may enhance the study of decision-making processes.</div></div><div><h3>New method</h3><div>We developed Lab-Life, a device enabling manipulation of real objects while ensuring precise monitoring and compatibility with electrophysiological recordings. Forty-four right-handed healthy volunteers performed two decision-making tasks: the Iowa Gambling Task (IGT, real vs. virtual cards) and the Game of Dice Task (GDT, real vs. virtual dice). Twenty-two participants (11 per task) used Lab-Life (hybrid condition), while additional virtual task groups were included to illustrate typical behavioral and EEG signatures. Object identity and values were tracked with infrared cameras, and EEG was recorded to analyze event-related potentials (ERPs) to outcomes.</div></div><div><h3>Results</h3><div>Behavioral analyses showed perfect concordance between expected and detected object values in hybrid condition, validating Lab-Life’s automated object recognition. EEG analyses revealed comparable numbers of valid trials and similar ERP patterns between hybrid and virtual task conditions, indicating that the device does not introduce movement artifacts. Participants consistently reported higher enjoyment when manipulating real compared to virtual objects.</div></div><div><h3>Comparison with existing methods</h3><div>Unlike conventional paradigms relying solely on virtual stimuli, Lab-Life integrates real objects without compromising behavioral or electrophysiological data quality. The device allows precise temporal synchronization between object manipulation and EEG recordings while preserving experimental control.</div></div><div><h3>Conclusions</h3><div>Lab-Life is a validated methodological tool for combining behavioral and electrophysiological measures with real object manipulation. It offers a flexible and adaptable platform for decision-making, memory, or perceptual tasks, thereby bridging the gap between laboratory experiments and real-life conditions. Larger studies are warranted to further explore its impact on cognitive performance.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110692"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146025589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research progress in vivo, in vitro, and in silico models of intracranial atherosclerotic stenosis","authors":"Fanqiang Meng ,&nbsp;Hanlu Xiang ,&nbsp;Mengjie Huo ,&nbsp;Jiaqi Wu ,&nbsp;Wentao Yao ,&nbsp;Hebo Wang","doi":"10.1016/j.jneumeth.2026.110685","DOIUrl":"10.1016/j.jneumeth.2026.110685","url":null,"abstract":"<div><div>Intracranial atherosclerotic stenosis (ICAS) is a major global cause of stroke with high recurrence risk. However, research on its pathophysiology is limited by difficulties in obtaining histopathological samples and the absence of adequate experimental models. The rising incidence of ICAS imposes a significant burden on families and healthcare systems, highlighting the urgent need for affordable, robust, and reliable animal models to study its prevention and treatment. To better understand the disease, researchers frequently employ animal models to replicate the progression of ICAS in humans. In recent years, many researchers have provided a variety of solutions for modeling methods and intervention protocols in animal models of ICAS, which has greatly improved the success rate of atherosclerosis models. Nevertheless, variations in intervention factors, material selection, and treatment methods have resulted in no consensus on the optimal animal model for ICAS. This review evaluates the strengths and limitations of existing in vivo、in vitro model and computational models of ICAS and the atherosclerotic process, aiming to inform better future models and drive novel stroke-reducing treatment strategies.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110685"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145998558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Longitudinal multimodal assessment of neuropathy in a porcine neuritis model","authors":"Ethan D. Griswold ,&nbsp;Matthew S. Zabriskie ,&nbsp;Lexie Scarpa ,&nbsp;Eric Goold ,&nbsp;Henrik Odeen ,&nbsp;Lubdha M. Shah ,&nbsp;Viola Rieke","doi":"10.1016/j.jneumeth.2026.110694","DOIUrl":"10.1016/j.jneumeth.2026.110694","url":null,"abstract":"<div><h3>Background</h3><div>Evaluation of animal models is critical in pre-clinical therapeutic research to determine translation potential. Due to their anatomic and physiological resemblance to humans, porcine models are a critical step prior to human translation. Our work evaluates long-term anatomic and behavioral effects (n = 9) over 8 weeks post-neuritis-induction surgery of the common peroneal nerve (CPN).</div></div><div><h3>New method</h3><div>This model utilizes longitudinal MRI and histological analysis of nerve and muscle, in addition to the use of the algometer and feather test as quantitative sensory testing methods.</div></div><div><h3>Results</h3><div>Neuritis animals showed heightened rear leg sensitivity on the neuritis side, with significant reduction in maximum tolerated pressure in the majority of dermatomes and significant heightened sensitivity to light touch stimuli. Significant muscle loss was well defined with short- and long-term assessments. MR imaging showed increased signal intensity in CPN-innervated muscles alongside reduction in volume and hypotonia. Histology supports these conclusions as evidenced by the presence of atrophied fibers, fibrosis, and fatty infiltrations. At necropsy, there was an average of 50 % reduction in CPN-innervated muscle weights compared to control leg muscles. Pigs experienced postural and ambulatory abnormalities without inhibiting regular activity.</div></div><div><h3>Comparison with existing method(s)</h3><div>We expand on established protocols for assessing neuropathic pain (Castel et al., 2016; Hellman et al., 2021b), which assess short-term effects (10 days – 4 weeks) of a peripheral nerve injury.</div></div><div><h3>Conclusion</h3><div>We have demonstrated and evaluated a longitudinal chronic neuropathic pain porcine model. Variables including medication regime and anatomic sequelae need to be considered as they may affect the quantitative sensory testing and time course of testing.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110694"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146010793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Micromap: A low-cost multi-channel electrophysiology acquisition system","authors":"João Pedro Carvalho-Moreira ,&nbsp;Luiz Felipe Nogueira Varandas ,&nbsp;Matheus Costa Passos ,&nbsp;Márcio Flávio Dutra Moraes","doi":"10.1016/j.jneumeth.2026.110691","DOIUrl":"10.1016/j.jneumeth.2026.110691","url":null,"abstract":"<div><h3>Objective</h3><div>To develop an accessible, low-cost, and open-source system for multi-channel electrophysiological recordings across multiple brain regions, facilitating the investigation of neural dynamics and information flow in long-range integration. Approach: We designed a digital signal acquisition system that integrates two programmable analog-to-digital converter chipsets -- Intan RHD and Texas Instruments ADS -- with an Arduino-based microcontroller. In addition, we introduce a custom-designed headstage and a methodology for spatially distributed electrode placement using a perforated printed circuit board, which interfaces directly with the system to enable local field potential recordings from multiple brain regions. The system successfully recorded biopotentials without significant sample loss and performed comparably to gold-standard systems, enabling recordings across 32 channels at a 2 kHz sampling frequency. We verified signal integrity and stability under both experimental and simulated conditions, confirming the platform’s suitability for electrophysiology. Main results: We present a comprehensive, low-cost bioengineering solution for distributed electrophysiological recordings, enabling detailed investigation of brain dynamics through synchronized neural activity across regions. Significance: This open-source platform enables neuroscience laboratories to investigate large-scale neural interactions with high temporal precision, expanding access to systems neuroscience tools essential for understanding interregional brain connectivity.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110691"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145994270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OpenIrisDPI: An open-source digital dual Purkinje image eye tracker for visual neuroscience","authors":"Ryan A. Ressmeyer ,&nbsp;Jorge Otero-Millan ,&nbsp;Gregory D. Horwitz ,&nbsp;Jacob L. Yates","doi":"10.1016/j.jneumeth.2026.110693","DOIUrl":"10.1016/j.jneumeth.2026.110693","url":null,"abstract":"<div><h3>Background</h3><div>Video-based eye trackers are widely used in vision science, psychology, clinical assessment, and neurophysiology. Many such systems track the pupil center and corneal reflection (P-CR) and compare their positions to estimate the direction of gaze. However, P-CR eye trackers are often too imprecise for applications with stringent eye tracking quality requirements.</div></div><div><h3>New method</h3><div>We present OpenIrisDPI, an open-source plugin for the OpenIris framework that implements dual Purkinje image (DPI) tracking. OpenIrisDPI supports simultaneous pupillography, a technique widely used in perceptual psychology and neuroscience, and it enables direct comparison between P-CR and DPI signals.</div></div><div><h3>Results</h3><div>Data collected from macaque monkeys using OpenIrisDPI show that the P-CR method overestimates the amount of fixational drift between saccades compared to DPI. The accuracy of the DPI signal was further validated using high-density extracellular recording of neurons in the lateral geniculate nucleus. Compensating for the effects of fixational eye movements using DPI signals produced sharper estimates of neuronal receptive fields than using simultaneously collected P-CR signals.</div></div><div><h3>Comparison with existing methods</h3><div>OpenIrisDPI is provided as open-source software and operates on consumer-grade hardware, making it more accessible than previously described DPI eye trackers and less costly than many P-CR systems. To our knowledge, OpenIrisDPI is the first eye tracker to perform both pupillography and DPI eye tracking.</div></div><div><h3>Conclusion</h3><div>OpenIrisDPI makes high-precision eye tracking readily available to the research community. It is well suited for visual neuroscience applications, where accurate knowledge of the retinal image during experiments is critical.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110693"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146046760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Artificial intelligence and machine learning driven nanorobotics for targeted brain delivery: Redefining blood-brain barrier navigation","authors":"Suraj Kumar ,&nbsp;Rishabha Malviya ,&nbsp;Sathvik Belagodu Sridhar ,&nbsp;Tarun Wadhwa ,&nbsp;Javedh Shareef","doi":"10.1016/j.jneumeth.2026.110695","DOIUrl":"10.1016/j.jneumeth.2026.110695","url":null,"abstract":"<div><div>The blood-brain barrier (BBB) plays a central role in preserving central nervous system (CNS) homeostasis, and its dysfunction is implicated in various neurological disorders and brain malignancies. To emulate the complex architecture and signaling environment of the BBB, researchers have developed advanced <em>in vitro</em> models incorporating three-dimensional (3D) cell cultures, organoid technologies, and microfluidic platforms. These innovations have significantly expanded the scope of neuropharmacological research, particularly in understanding BBB transport mechanisms and enhancing drug delivery strategies. The following review highlights the integration of biomolecular tools and nanomaterial-based engineering approaches to facilitate the targeted delivery of theranostic agents across the BBB. Specifically, it discusses the emerging role of nanorobots and nanosystems in mediating cellular interactions that enable precise modulation and penetration of the barrier. Case studies addressing therapeutic applications in brain tumors and cognitive dysfunctions are also evaluated. Moreover, the review explores the intersection of artificial intelligence (AI), machine learning (ML), and robotics, which together offer transformative potential in designing intelligent nanorobots capable of efficient and safe BBB traversal. Collectively, this work serves as a consolidated resource for clinicians, biomedical scientists, and technologists aiming to develop innovative strategies for BBB-targeted therapies and biopharmaceutical applications.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110695"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146025588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in rodent oligodendrocyte precursor cells isolation and culture: From traditional methods to modern approaches","authors":"Hoda Akbari,&nbsp;Iraj Ragerdi Kashani,&nbsp;Farzaneh Rezaei Yazdi,&nbsp;Parichehr Pasbakhsh,&nbsp;Sina Mojaverrostami","doi":"10.1016/j.jneumeth.2026.110706","DOIUrl":"10.1016/j.jneumeth.2026.110706","url":null,"abstract":"<div><h3>Introduction</h3><div>Oligodendrocyte progenitor cells (OPCs) are responsible for the generation of myelinating oligodendrocytes and are essential for myelination and repair of the central nervous system (CNS). In past decades, many methods have been presented for the reliable isolation and culture of OPCs to facilitate research into oligodendroglial biology and the development of effective therapeutic strategies such as cell transplantation and drug therapy in demyelinating and neurodegenerative diseases. However, challenges of purity, yield, and phenotypic stability still persist for each method.</div></div><div><h3>Methods</h3><div>A literature review was conducted using related keywords in the Scopus, Google Scholar, PubMed, and PubMed/Medline databases from the date of beginning until 1 November 2025. We examined preclinical research in rodent models that fit our search parameters, including in vitro and in vivo rat or mouse models, for both neonatal and adult CNS studies.</div></div><div><h3>Results</h3><div>This review systematically describes the methodological progress of OPC isolation from mechanical isolation methods to advanced immunopanning culture systems, magnetic-activated cell sorting (MACS), and serum-free or growth factor-based culture systems. Additionally, it mentions important novel methods, like 3D or normoxic culture models, that have improved cell purity, function, and physiological relevance.</div></div><div><h3>Discussion</h3><div>This article compares the principles, advantages, and disadvantages of each method to create a single set of guidelines for selecting the best methods regarding the equipment and goals of an investigation. It also suggests ways to design OPC isolation systems that are more standardized, scalable, and ethically acceptable, which may accelerate in the use of these systems in regenerative neuroscience.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110706"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146157406","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel transgenic reporter mouse line of astrocyte-derived small extracellular vesicles for investigating intercellular communication in vivo","authors":"Zhongwu Liu ,&nbsp;Yi Zhang ,&nbsp;Amy Kemper ,&nbsp;Yanfeng Li ,&nbsp;Andrew C. Dudley ,&nbsp;Michael Chopp ,&nbsp;Zheng Gang Zhang","doi":"10.1016/j.jneumeth.2026.110705","DOIUrl":"10.1016/j.jneumeth.2026.110705","url":null,"abstract":"<div><h3>Background</h3><div>Small extracellular vesicles (sEVs) mediate intercellular communication in the central nervous system, regulating processes ranging from homeostatic maintenance to injury repair. Although astrocytes are a major source of sEVs in the brain, in vivo investigation of their endogenous and cell-specific signaling remains technically challenging.</div></div><div><h3>New method</h3><div>To address this limitation, we developed a novel inducible transgenic reporter mouse line, GFAP-CD63-GFP, that enables specific labeling of astrocyte-derived sEVs. The mouse line was generated by crossing GFAP-CreERT2 mice with CD63-emGFPloxP/stop/loxP mice. This system enables Tamoxifen-inducible, astrocyte-specific expression of GFP-tagged CD63, a tetraspanin enriched in sEVs. The model allows visualization and quantification of astrocyte-derived CD63-positive sEVs in vivo.</div></div><div><h3>Results</h3><div>Following Tamoxifen induction, GFP expression was robustly detected in the brain and spinal cord. Immunogold transmission electron microscopy further identified GFP-positive sEVs within neurons, providing ultrastructural evidence of astrocyte-to-neuron vesicle transfer. As a proof-of-concept, ischemic stroke significantly increased astrocyte-derived sEVs in the ipsilesional cerebral hemisphere and the stroke-impaired side of the spinal cord, accompanied by enhanced neuronal endocytosis.</div></div><div><h3>Comparison with existing methods</h3><div>Current approaches rely primarily on in vitro EV isolation or nonspecific membrane dyes. The GFAP-CD63-GFP model enables cell type–specific, temporally controlled, and in situ tracking of astrocyte-derived sEVs.</div></div><div><h3>Conclusions</h3><div>These findings provide the first in vivo demonstration of increased astrocyte-to-neuron sEV communication during post-stroke recovery. The GFAP-CD63-GFP reporter mouse thus provides a powerful platform for investigating astrocyte-derived sEV signaling under both physiological and pathophysiological conditions of the CNS.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"429 ","pages":"Article 110705"},"PeriodicalIF":2.3,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}