神经病理学

1. Cell Line Preparation

Thaw approximately 1,000,000 mouse neuroblastoma spinal cord cells (NSC-34 cells) that were stored in a liquid nitrogen ...

1. Fluorescent Labeling and Immunocytochemistry
Note: The immunofluorescent labeling of primary cortical cell cultures was carried out in glass bottom 35 ...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Tissue harvesting and ...

1. Experimental Preparation
NOTE: The following stock solutions can be prepared and maintained until use.

Dissection Solution

Dissolve 3.62 g of ...

1. Microglial Cell Culture Maintenance

Warm cell culture medium (Dulbecco's Modified Eagle Medium; DMEM) supplemented with 10% fetal bovine serum ...

1. Preparation of Reagents and Equipment

Prepare 1x Dissection Buffer (70 mM NaCl, 5 mM KCl, 0.02 mM CaCl2, 20 mM MgCl2, 10 mM NaHCO3, 115 mM sucrose, ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

1. Maintenance and culture of undifferentiated human embryonic stem cells (hESCs)

Perform maintenance and expansion of hSECs on feeder-free conditions ...

1. hESC-derived dopaminergic organoids for Parkinson's Disease (PD) studies

Day 0: Amplify hESCs in 2D culture up to 60% confluency (day 0), then ...

1. Collapse Assay

Poly-D-lysine coating

Coat 8-well culture slides with 400 μL of 5 μg/mL poly-D-lysine (PDL) in phosphate-buffered ...

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.  
1. Preparation of Tissue ...

1. Neurons Treatment
Note: The treatment has been performed at Division (DIV) 7. All the steps are carried on under a cell culture hood in sterile ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review ...

1. Preparation of Aβ42 Peptide

Synthesize peptides (see Table 1) using the standard 9-fluorenyl methoxycarbonyl (Fmoc) chemistry with ...

1. Stimulation and Measuring Odorant Receptor (OR) Activity Using the Real-Time cAMP Assay

Observe the transfected cells under a phase-contrast ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.  ...

1. Monitoring Transcellular Spreading of Prion-like Proteins By In Vivo Time-lapse Imaging
NOTE: Grow C. elegans wild-type (WT) (N2) and transgenic lines ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.  ...

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines.  
1. Homogenization and ...

1. Coupling Gal4- and QF-mediated Htt Transgene Expression in Drosophila

Collect and/or generate transgenic Drosophila melanogaster lines containing ...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Immunofluorescence
...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Preparation

Prepare ...

1. Proteasome Activity Assay
NOTE: Proteasome activity can be measured in homogenized brain tissue using a slightly modified version of the 20S Proteasome ...

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 
1. ...

Source: Donertas-Ayaz, B. et al., Assessment of Nerve Injury-Induced Mechanical Hypersensitivity in Rats Using an Orofacial Operant Pain Assay. J. Vis. ...

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Source: Pang, X., et al. A Stably Established Two-Point Injection of Lysophosphatidylcholine-Induced Focal Demyelination Model in Mice. J. Vis. Exp. ...

Swietek, B., et al., Immunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers. J. Vis. Exp. (2016)
This video demonstrates the ...

Source: Chi, S., Fang, et al. Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein ...

Source: Keough, M. B., et al. Experimental Demyelination and Remyelination of Murine Spinal Cord by Focal Injection of Lysolecithin. J. Vis. Exp. (2015)
...

Source: Aulston, B., et al. An In Vitro Model for Studying Tau Aggregation Using Lentiviral-mediated Transduction of Human Neurons. 
This video ...

Source: Liang, K. X., et al. Flow Cytometric Analysis of Multiple Mitochondrial Parameters in Human Induced Pluripotent Stem Cells and Their Neural and ...

Source: Wang, B., et al., Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase ...

Source: Schmidt, K. et al., Localization of the Locus Coeruleus in the Mouse Brain. J. Vis. Exp. (2019)
This protocol outlines the use of ...

Source: Yamada, K. In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off ...

Source: Son, H., et al., A Chronic Immobilization Stress Protocol for Inducing Depression-Like Behavior in Mice. J. Vis. Exp. (2019).
This video ...

Source: Brai, E., et al. An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices. J. Vis. Exp. (2018)
This ...

Source: Lin, Y., et al., Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release. J. Vis. Exp. (2018)
This video demonstrates ...

Source: Singh, P., et al. Stereological Estimation of Cholinergic Fiber Length in the Nucleus Basalis of Meynert of the Mouse Brain. J. Vis. Exp. (2020)
...

Source: Jensen, K. L., et al., Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal ...

Source: Economides, G., et al. Biocytin Recovery and 3D Reconstructions of Filled Hippocampal CA2 Interneurons. J. Vis. Exp. (2018)
This video ...

Source: Shokry, I. M., et al. Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin ...

Source: Yamaguchi, K., et al.  Assessment of Long-term Depression Induction in Adult Cerebellar Slices. J. Vis. Exp. (2019)
In this video, a mouse ...

Source: Batelli, S., et al. Application of MultiColor FlpOut Technique to Study High Resolution Single Cell Morphologies and Cell Interactions of Glia in ...

Source: Lamas, V., et al. Stereotactically-guided Ablation of the Rat Auditory Cortex, and Localization of the Lesion in the Brain. J. Vis. Exp. ...

Source:  Le, T. P., et al., An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells. J. Vis. Exp. (2021)
In this ...

Source: Sumitomo, A., et al. BS3 Chemical Crosslinking Assay: Evaluating the Effect of Chronic Stress on Cell Surface GABAA Receptor Presentation in the ...

Source: Loganathan, S., et al., Measuring Glucose Uptake in Drosophila Models of TDP-43 Proteinopathy. J. Vis. Exp. (2021).
The video demonstrates a ...

Source: Eaker, A. R., et al. Analysis of Astrocyte Territory Volume and Tiling in Thick Free-Floating Tissue Sections. J. Vis. Exp. (2022)
This video ...

Source: Martuscello, R. T., L. et al., A Stainless Protocol for High Quality RNA Isolation from Laser Capture Microdissected Purkinje Cells in the Human ...

Source: Wager-Miller, J. et al.  Collection of Frozen Rodent Brain Regions for Downstream Analyses. J. Vis. Exp (2020)
This video demonstrates a ...

Source: Saravi, B. et al., A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease. J. Vis. Exp. ...

Source: Rubio, F. J., et. al. Fluorescence Activated Cell Sorting (FACS) and Gene Expression Analysis of Fos-expressing Neurons from Fresh and Frozen Rat ...

Source: Speca, D. J., et al. Acyl-PEGyl Exchange Gel Shift Assay for Quantitative Determination of Palmitoylation of Brain Membrane Proteins. J. Vis. Exp. ...

Source: Almari, B., et al. Fabrication of Amyloid-β-Secreting Alginate Microbeads for Use in Modelling Alzheimer's Disease. J. Vis. Exp. (2019).
...

Source: Iacono, D., et.al., Symmetric Bihemispheric Postmortem Brain Cutting to Study Healthy and Pathological Brain Conditions in Humans. J. Vis. Exp. ...

Source: Li, W., et. al. Mitochondrial Ca2+ Retention Capacity Assay and Ca2+-triggered Mitochondrial Swelling Assay. J. Vis. Exp. (2018)
This video ...

Source: Ajoy, R., et. al. Studying the Hypothalamic Insulin Signal to Peripheral Glucose Intolerance with a Continuous Drug Infusion System into the Mouse ...

Source: Bunker, S., et al. Experimental glaucoma induced by ocular injection of magnetic microspheres. J. Vis. Exp. (2015)
In this video, an anesthetized ...

Source: Kummari, E., et. al., Laser Capture Microdissection - A Demonstration of the Isolation of Individual Dopamine Neurons and the Entire Ventral ...

Source: Bennett, S. A., et al., Characterizing Histone Post-translational Modification Alterations in Yeast Neurodegenerative Proteinopathy Models. ...

Source: Yan, Z., et al. Evaluating the Anti-depression Effect of Xiaoyaosan on Chronically-stressed Mice. J. Vis. Exp. (2019).The video demonstrates the ...

Source: Bétemps, D., et al.,  Detection of Disease-associated α-synuclein by Enhanced ELISA in the Brain of Transgenic Mice ...

Source: Wulf, M., et.al. Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules. J. Vis. Exp. ...

Source: Zhi, L. et al., Measurement of Oxygen Consumption Rate in Acute Striatal Slices from Adult Mice. J. Vis. Exp. (2022).This video demonstrates how ...

Source: Josiah, S. S., et al. Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting. J. Vis. Exp. (2022).The ...

Source: Lani-Louzada, R., et al.  Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents. J. Vis. Exp. ...

Source: Jensen, V. N., et al. Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease. J. Vis. ...

Source: Jiang, X., et al. Inducement and evaluation of a murine model of experimental myopia. J. Vis. Exp. (2019)This video demonstrates a procedure for ...

Source: Jolly, S., et.al. Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse ...

Source: Baleriola, J., et. al., Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization. J. Vis. ...

Source: Zhang, J., et al. High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses. J. Vis. Exp. (2016)This video ...

Source: de March, C. A. et. al., Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase. J. Vis. Exp. (2019)This ...

Source: Alluri, H., et al. Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier ...

Source: Eid, L. & Parent, M. Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy. J. ...

Source: Evilsizor, M. N., et. al., Primer for Immunohistochemistry on Cryosectioned Rat Brain Tissue: Example Staining for Microglia and Neurons. J. Vis. ...

Source: Kirchner, K.N., et al., A Hydrophobic Tissue Clearing Method for Rat Brain Tissue. J. Vis. Exp. (2020).This video demonstrates the ...

为了检测这些细菌，取经过脱蜡、复水处理的、感染细菌的人脑组织切片，该切片应暴露蛋白质抗原并已灭活内源性磷酸酶。...

该病毒可感染大脑皮层中的锥体神经元，表达的病毒蛋白定位于细胞质内称为包涵体的颗粒状结构中。...

洗涤以去除石蜡，然后使用浓度递减的酒精对组织进行再水化。...

本视频演示了从星形胶质细胞培养物中分离细胞表面蛋白和细胞内蛋白的方法。将星形胶质细胞培养物置于冰上以抑制内吞作用，随后用生物素化试剂标记细胞表面蛋白。细胞裂解后释放出生物素标记的细胞表面蛋白和未标记的细胞内蛋白。最后，使用链霉亲和素包被的磁珠将生物素标记的细胞表面蛋白与细胞内蛋白分离。...

本视频演示了脑室内注射以研究小鼠巨细胞病毒（MCMV）在新生小鼠脑组织中的分布。经基因工程改造后可表达增强型绿色荧光蛋白（EGFP）以便观察的MCMV，被注射至幼鼠脑的侧脑室。注射后的病毒通过脑脊液循环，感染脑室周围区域的细胞，包括边缘带、脉络丛和室下区。...

本视频展示了在发育中的大脑中建立寨卡病毒感染体外模型的方法。利用包含干细胞、神经前体细胞（NPCs）和神经元的玫瑰花结样结构的人脑类器官，进行寨卡病毒感染。该病毒可感染神经前体细胞并诱发细胞凋亡，从而破坏类器官的正常发育。...

该视频演示了将脂多糖（LPS）显微注射到斑马鱼幼鱼脑内以诱导神经炎症的过程。将幼鱼置于铺有琼脂糖的培养皿中，将注射针插入脑室，注入LPS。注入的LPS会触发免疫反应，从而导致神经炎症。...

本视频展示了在基因修饰的星形胶质细胞中评估线粒体健康状况的方法，这些细胞在线粒体内表达一种荧光蛋白。该蛋白的荧光会随线粒体健康状态的变化由绿色转变为红色，从而指示应激或损伤。通过多种参数对获取的图像进行捕获和分析。...

本视频演示了将嵌合型HIV病毒注射到大鼠脑皮层的操作步骤，其中病毒RNA会整合到宿主基因组中，进行复制并扩散，从而在实验模型中建立活跃的HIV感染。...

本视频展示了构建中枢神经系统感染小鼠模型的方法。通过颅内和腹腔联合注射，将病毒接种于 suckling...

本视频展示了在小鼠模型中诱导神经炎症的过程。取一只已麻醉并制造微小伤口的小鼠，将伪狂犬病病毒（PRV）接种于伤口处。病毒侵染外周神经，扩散至背根神经节（DRG），并进一步侵染脊髓和脑组织。神经系统中神经元的感染引发免疫反应，导致神经元损伤，从而诱发神经炎症。...

该视频展示了在成年小鼠脑内注射寨卡病毒悬液以诱导神经退行性变的过程。将麻醉后的小鼠固定于立体定位仪中，于暴露的颅骨上钻孔以便注射。将寨卡病毒悬液注入脑组织中。病毒感染引发炎症反应，进而导致神经退行性变。...

本视频展示了从感染并经灌流的小鼠脑组织中定量可培养细菌载量的方法 单核细胞增生李斯特菌.

本视频展示了一种利用膜插件上培养的星形胶质细胞和内皮细胞构建的体外血脑屏障（BBB）模型来研究物质渗透的方法，实验中使用了游离的FITC和负载FITC的铁蛋白。FITC标记的铁蛋白可与内皮细胞表面的受体结合，并穿过膜屏障进入下室，而游离FITC的渗透受到限制，主要保留在上室。...

本视频展示了一种用寨卡病毒感染人类胎儿神经干细胞的操作流程。感染后的细胞可用于进一步的研究。...

本视频展示了一种用于研究病原体穿越血脑屏障（BBB）的体外模型。表达紧密连接的微血管内皮细胞在上室形成内皮层，而神经元、星形胶质细胞和小胶质细胞则在下室代表脑实质，两者之间由多孔膜隔开。在该BBB模型中进行病毒感染，可模拟病原体从血液进入脑组织的过程。当引入嗜神经病毒后，病毒通过转胞吞作用穿越血脑屏障，感染神经细胞，进行复制并释放子代病毒，从而展示出成功的神经侵入过程。...

来源：Funes, S., 等.，人类小胶质样细胞：由诱导性多能干细胞分化及利用人突触体的体外活细胞吞噬实验。视频实验杂志...

将组织浸入二甲苯中以溶解石蜡。...

静脉注射荧光标记的白蛋白，该白蛋白可穿过受损的血脑屏障进入脑组织。...

该组织显示出脑血管壁中的淀粉样纤维和异常蛋白质聚集体，这是神经系统疾病的關鍵指標。...

MCCH...

将细胞混合物悬浮于细胞外基质（ECM）溶液中。...

做颈部切口以暴露双侧颈总动脉，并进行松扎。...

本视频演示了通过注射髓鞘来源的多肽并联合佐剂以激活自身反应性T细胞来诱导实验性自身免疫性脑脊髓炎（EAE）的方法。这些T细胞会迁移至脑部。同时给予百日咳毒素以增加血脑屏障的通透性，使这些T细胞及其他免疫细胞得以进入脑组织。这些免疫细胞引发脱髓鞘，导致神经传导受损，从而诱发EAE。...

本视频演示了在实验性自身免疫性脑脊髓炎（EAE）小鼠模型中评估血脑屏障破坏的实验方法。通过眼眶后注射不同分子量的荧光染料，使其在脑部血管中循环。较小分子量的染料可渗漏进入脑组织，而较大分子量的染料仅在血脑屏障严重受损时才能进入，从而反映屏障损伤的程度。...

本视频演示了通过电刺激在麻醉大鼠的脑桥中诱导组织损伤或梗死的操作步骤。...

在本视频中，受精的斑马鱼胚胎在体视显微镜下进行去壳处理，随后暴露于胆固醇合成抑制剂中，导致幼鱼出现血管破裂和脑出血。通过显微镜观察，可根据明显的红色斑点识别出血性幼鱼，并将其分离以进行后续分析。...

本视频展示了在大鼠模型中使用脂多糖（LPS）诱导血脑屏障（BBB）破坏及脑微出血的过程。重点阐述了导致血脑屏障通透性增加及继发性脑组织损伤的免疫反应序列。...

本视频展示了一种在大鼠模型中检测脑微出血（CMHs）的实验方法。CMHs 是由于血脑屏障破坏导致的脑内出血，通过外渗的红细胞（RBCs）进行识别。在大鼠模型中，固定后的脑组织经冷冻保护、切片，并用苏木精-伊红染色以标记红细胞，从而可在显微镜下观察...

该视频演示了建立小鼠模型以研究新生儿缺氧缺血性脑损伤的操作流程。视频概述了通过手术诱导缺血，并使动物暴露于低氧环境以加重脑损伤的各个步骤。最后，展示了如何评估所形成的脑部病变，确保该小鼠模型可用于后续科学研究。...

本视频演示了如何利用离子选择性微电极和电泳微电极测量并比较电信号，以表征脑切片中的细胞外间隙。该过程有助于确定细胞外间隙的体积分数及其迂曲程度。...

在本视频中，将一个涂有谷氨酸氧化酶的多电极阵列（MEA）植入麻醉小鼠的脑组织中。将谷氨酸溶液注入细胞外空间，MEA...

在本视频中，使用了一只经过基因改造的幼鼠，其Ccm2基因两侧带有loxP位点，并融合了雌激素受体的Cre重组酶，用于诱导脑海绵状血管畸形（CCMs）。通过给予4-羟基他莫昔芬激活内皮细胞中的Cre，导致Ccm2基因被删除，从而在脑内形成CCMs。...

本视频演示了通过酶消化法从小鼠缺血脑组织中分离脑细胞和浸润的免疫细胞的实验步骤，随后利用密度梯度离心去除细胞碎片和髓鞘，以用于脑卒中研究中的炎症反应分析。...

本视频展示了通过血管内穿刺法在小鼠模型中诱导蛛网膜下腔出血的操作过程。对麻醉后的小鼠颈部区域进行切口，以暴露颈总动脉及其分叉部位。将一根细丝经动脉插入，穿刺颅内动脉分支，从而诱导蛛网膜下腔出血。...

本视频展示了一种建立新生大鼠脑室内出血模型的方法。在该操作中，向麻醉后的大鼠幼崽脑侧脑室注射血红蛋白。血红蛋白可引起氧化应激并释放血红素，从而模拟脑室内出血。由此产生的脑组织损伤由活性氧和炎性细胞因子介导，导致脑室扩大，这是脑室内出血的常见后果。...

本视频演示了在老年大鼠中进行远端大脑中动脉永久性阻塞（MCAO）并结合颈总动脉（CCA）结扎以模拟缺血性脑卒中的操作步骤。该方法可阻断脑部血流，导致神经元死亡，从而建立用于研究脑卒中机制及潜在治疗方法的动物模型。...

本视频演示了建立蛛网膜下腔出血大鼠模型的操作步骤。对麻醉后的大鼠植入探针，用于测量颅内压和脑血流量。确定目标位置后，将针头插入视交叉前位置。随后将自体血注入蛛网膜下腔，导致颅内压升高、脑血流量下降，从而成功建立模型。...

本视频展示了一项关于淋巴细胞穿过血脑屏障（BBB）内皮模型的实验研究。在体外培养形成一层人脑微血管内皮细胞（HBMECs）单层，随后将含有淋巴细胞的外周血单个核细胞（PBMCs）加入该单层细胞中。通过孵育使淋巴细胞发生跨内皮迁移，迁移后的细胞通过流式细胞术进行定量分析。...

本视频演示了如何评估经大脑中动脉阻塞处理的大鼠脑切片中的梗死区域和脑水肿。使用氧化还原染色法识别梗死区域，同时通过图像分析软件中的蓝色滤光片和阈值功能，对受损半脑的梗死区域和水肿进行定量分析。...

本视频展示了一种在体外评估小鼠脑微血管内皮细胞跨内皮电阻（TEER）的方法。较高的TEER值表明内皮细胞连接完整，而加入活化的T细胞后TEER值降低，反映出连接屏障的破坏。...

本视频演示了在血管紧张素II诱导的高血压小鼠模型中，采用苯酚破坏肾动脉交感神经功能以降低血压的操作步骤。...

本视频演示了通过使用栓线阻塞大脑中动脉（MCA）在大鼠模型中诱导缺血性脑卒中的过程。该操作包括结扎颈动脉，将栓线插入颈外动脉，并阻断大脑中动脉向脑部的血流，从而导致神经元死亡，模拟缺血性脑卒中。...

本视频展示了通过阻断小鼠右侧颈总动脉并将其暴露于低氧环境中，在小鼠模型中复制血栓性中风的操作过程。该过程会减少向大脑的血液和氧气供应，导致血栓形成及脑损伤。...

本视频展示了通过向麻醉小鼠脑白质内特定脑区坐标注射一氧化氮抑制剂，以精确诱导白质脑卒中的技术。该方法可导致血管收缩并阻断血液供应，从而引起神经元损伤和脑卒中。...

来源：Wanderer, S., 等。兔动脉囊微血管吻合口动脉瘤模型。J. Vis. Exp.

来源：Sharma, T. P.，等，用于调节人供体眼球后段眼内压与颅内压的跨层自主系统模型。J. Vis. Exp.

来源：Lani-Louzada,...

来源：Jungen, C., 等心脏内神经元对心脏电生理学及心律失常发生的影响 离体 朗根多夫系统 视频实验杂志...

来源：Hakim, M. A. 等，小鼠脑实质微动脉内皮的分离与功能分析。J. Vis. Exp.

来源：Tang, X. 等. 通过自体血双次注射建立桥脑大量出血模型. J. Vis. Exp.

来源：Alluri, H., 等. 氧糖剥夺与再氧合作为一种 体外 用于研究血脑屏障功能障碍的缺血-再灌注损伤模型 J. Vis. Exp.

来源：Gucciardo, E., 等.，增生性糖尿病视网膜病变中纤维血管并发症的离体组织培养模型。J. Vis. Exp.

来源：Talley Watts,...

来源：Kuts, R. 等．大鼠运动皮层激光诱导脑损伤．J. Vis. Exp.

1. Chemo-attraction Assay

Place a three-well culture insert in the center of a poly-L-ornithine/laminin-coated 35 mm dish using sterile tweezers.
Turn ...

1. Immunostaining of organoid slices

Place the microscope slides containing organoid sections in a copling jar with 50 mL of prewarmed 1x PBS, holding ...

1. FDA-PI Double Staining and in Cerebellar Granule Neuron (CGN) Culture

Prepare fluorescein diacetate-propidium iodide (FDA-PI) working solution by ...

1. Fixation and Immunofluorescent Staining Procedure

Using a p200 pipette, transfer dissected brains from the 9-well dish to a 0.5 mL microcentrifuge ...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Immunohistochemistry

...

Disclaimer text: All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.   ...

1. Imaging Vesicle Motility

Prepare the plasmids that will label each type of vesicle. For instance, use EGFP-Rab5 for early endosomes, EGFP-Rab7 for ...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Solution and Media ...

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Immunostaining Protocol

...

Source: Vogt, D. et al., Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies. J. Vis. ...

Source: Klein, B. Y. et al., Assessing Cellular Stress and Inflammation in Discrete Oxytocin-secreting Brain Nuclei in the Neonatal Rat Before and After ...

Source: Pericoli, G. et. al., Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma. J. Vis. Exp. (2021)This video demonstrates live ...

Source: Jantzie, L. L., et. al. Modeling Encephalopathy of Prematurity Using Prenatal Hypoxia-ischemia with Intra-amniotic Lipopolysaccharide in Rats. J. ...

Source: Larpthaveesarp, A., et. al. Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats. J. Vis. Exp. (2017)This video ...

该疾病会触发自身反应性免疫细胞攻击髓鞘蛋白，导致神经元轴突周围的髓鞘降解。...

对照组切片显示轴突具有紧密排列的髓鞘，其中包含同心圆状的质膜层。损伤组切片则显示髓鞘结构松散，出现较大间隙。...

应激会损害少突胶质细胞功能，破坏内侧前额叶皮层神经元轴突周围富含脂质的髓鞘结构，而内侧前额叶皮层是调控情绪的脑区。...

本视频展示了利用靶蛋白特异性抗体包被的磁珠，从干细胞来源的运动神经元裂解液中免疫沉淀与靶蛋白交联的DNA片段。...

本视频展示了一种用于评估背侧纵肌中神经肌肉接头结构完整性的实验方案...

本视频展示了利用荧光标记神经元评估测试化学品对秀丽隐杆线虫（Caenorhabditis...

本视频演示了一种利用TRPV1激动剂树脂毒素（resiniferatoxin,...

在本视频中，采用实验性自身免疫性脑脊髓炎（EAE）建立小鼠多发性硬化模型。将含有神经元肽和佐剂的乳剂注射至小鼠体内，引发免疫反应，激活自身反应性T细胞。随后注射百日咳毒素（PTX）以增加血脑屏障通透性，使自身反应性T细胞得以浸润脑组织。此类浸润启动信号级联反应，最终导致髓鞘损伤，模拟多发性硬化症的脱髓鞘病变状态。...

本视频展示了通过将靶向水通道蛋白-4的活化Th17细胞转移至小鼠模型来诱导瘫痪的操作过程。在中枢神经系统中，活化的Th17细胞与表达水通道蛋白-4的星形胶质细胞相互作用，引发免疫反应，导致小鼠出现尾部和肢体瘫痪。...

本视频展示了通过注射百日咳毒素以及含有模拟髓鞘肽段和佐剂的接种物，在小鼠模型中评估运动功能障碍的过程，随后进行跑台实验以评估运动功能，并评价因免疫激活和脱髓鞘所导致的神经损伤。...

本视频演示了阶梯实验，用于评估右侧大脑损伤的偏侧帕金森病大鼠的运动功能。该损伤会破坏多巴胺信号传导，导致左侧前爪控制能力受损，从而使得左侧阶梯上残留更多的食丸，表明存在运动功能障碍。...

本视频演示了使用径向运动实验评估表达野生型或ALS突变型人类TDP-43蛋白的C. elegans中运动功能障碍的方法。让表达不同水平野生型或突变型TDP-43的线虫自由爬行，并通过其在实验平板上从中心点出发所移动的径向距离来测量其运动能力。该实验根据不同C.

本视频展示了利用培养的运动神经元、施万细胞和肌管评估肌肉功能，随后使用神经肌肉阻滞剂抑制收缩。彩色编码的时间-运动图像显示了彩色编码的时间-运动图示，其中红色表示最快的运动，蓝色表示最慢的运动。...

本视频展示了利用邻近连接实验（Proximity Ligation Assay,...

来源：Wagner, K. 等，利用eGFP标记蛋白研究果蝇感光细胞中的膜蛋白转运。J. Vis. Exp.

来源：Deseure, K., 等. 通过大鼠眶下神经慢性缩窄性损伤（IoN-CCI）研究三叉神经病理性疼痛。J. Vis. Exp.

来源：Brakkee, E. M., 等 大鼠胫骨神经瘤移位模型中的手术操作与行为学检测 视频实验杂志...

来源：Lee, D. et. al., 小鼠噪声性听力损失的改良实验条件及听力功能与外毛细胞损伤评估。J. Vis. Exp.

来源：Lee, D., 等．小鼠噪声性听力损失的改良实验条件及听力功能与外毛细胞损伤评估．J. Vis.

来源：Olmstead, D. N., 等，小鼠面神经轴突切断模型：研究运动神经元对损伤反应的方法。J. Vis. Exp. (2015)。本视频演示了小鼠面神经 crush...

来源：Cheah,...

为了使用TUNEL法检测凋亡性细胞死亡，用蛋白酶K处理脑切片。...

利用荧光显微镜观察受损神经，可见其轴突肿胀和断裂现象逐渐增加。...

药物包封可增强对肿瘤细胞的特异性，并降低神经毒性。...

本视频展示了在小鼠中实施创伤性脑损伤（TBI）的方法。暴露颅骨后，标记TBI作用部位。将小鼠置于TBI装置下方，通过提起金属杆并使其自由下落撞击与颅骨接触的冲击头，从而产生冲击损伤。...

本视频演示了在微流控芯片内培养的大鼠海马神经元的免疫荧光染色，用于分析神经元的成熟与连接性。...

本视频演示了在小鼠原代小脑颗粒神经元中建立神经元损伤模型的方法。该方法概述了使用NMDA（一种兴奋性神经递质受体激动剂）和甘氨酸（一种共激动剂）诱导神经元损伤的步骤。NMDA和甘氨酸结合于特定的神经元受体，导致受体过度激活，从而引起大量细胞内钙离子内流，最终导致神经元损伤和死亡。...

本视频演示了对小鼠脑切片进行免疫荧光染色，以定量分析特定突触前蛋白的分布情况。该过程包括封闭非特异性结合位点，依次加入靶蛋白的一抗和参考标记物的一抗，随后进行二抗标记以及细胞核复染。参考标记物可提供一致的基准信号，确保定量结果的准确性，通过共聚焦显微镜对突触前蛋白在不同脑区的分布进行分析。...

本视频演示了一种利用大规模扫描透射电子显微镜（STEM）研究斑马鱼幼体脑损伤的方法。STEM技术结合了扫描电子显微镜（SEM）和透射电子显微镜（TEM）的特点，可生成高分辨率图像。本文概述了样品制备的步骤以及高分辨率STEM图像的获取过程，以用于识别脑损伤。...

本视频演示了在大鼠模型上进行的外科手术方案，详细介绍了从麻醉准备到面神经损伤诱导的各个步骤。该方案在医学研究中对于研究神经再生和外科技术具有重要意义。...

本视频演示了通过开颅方式在小鼠中诱导创伤性脑损伤的操作步骤：沿颅骨中线做切口，钻透颅骨并移除骨瓣以暴露脑皮层，施加高压以造成损伤。随后缝合伤口，制备可用于后续实验的小鼠模型。...

本视频演示了通过球囊导管可控地增加颅内压，从而在小鼠模型中诱导脑死亡的实验操作，以模拟自然状态下的脑死亡过程。...

本视频演示了一种利用激光在大鼠运动皮层产生精确、可控脑损伤的操作方法。...

本视频展示了在原代神经干细胞和祖细胞以及神经元中，对修饰的唾液酸糖蛋白进行生物素化和荧光标记，并利用荧光显微镜进行可视化观察。...

在本视频中，通过受控加压使聚合物隔膜破裂，利用激波管装置产生激波。激波作用于固定在密封袋中的脑切片，从而诱导脑切片发生创伤性损伤。...

本视频演示了为光学分析回收突触小泡而制备培养神经元的实验步骤。该过程包括对突触小泡蛋白进行荧光标记以及通过免疫染色来标记和可视化神经元内的回收小泡。...

本视频展示了在人诱导性多能干细胞来源的神经元（hiPSCNs）上诱导拉伸损伤的过程。将细胞接种于底部无底的多孔板中，该板带有附着的硅胶膜。通过对膜施加可控的机械应力，引起细胞骨架破坏和细胞膜损伤，从而在细胞中诱导出拉伸损伤表型。...

本视频演示了一种使用气体驱动撞击装置对麻醉的黑腹果蝇（Drosophila）造成闭合性头部创伤性损伤的操作步骤。该损伤会影响神经元功能，并损害果蝇的行走能力。...

本视频演示了小鼠皮层刺伤损伤的操作过程，包括颅骨暴露和脑损伤的建立。该模型可诱导反应性星形胶质细胞增生，即星形胶质细胞的一种神经保护性反应，用于研究中枢神经系统创伤、炎症及修复机制。...

该视频演示了一种使用侧向液体脉冲装置（LFPI）建立小鼠创伤性脑损伤（TBI）模型的方法。在对小鼠实施麻醉后，通过在颅骨上开窗暴露脑组织。将充满生理盐水的套管连接至颅骨窗，并与LFPI装置相连。释放装置的摆锤以产生压力脉冲，从而造成创伤性脑损伤。...

本视频演示了在小鼠模型中诱导创伤性脑损伤（TBI）的操作步骤。将一只麻醉后暴露颅骨的小鼠置于操作台，通过颅骨开窗，使用撞击装置诱导可控的皮层损伤。该损伤可破坏脑组织，导致认知功能障碍和运动协调能力受损。...

本视频演示了在大鼠中进行胸段脊髓半切手术的操作步骤。内容包括暴露T8脊髓节段、实施单侧脊髓半切以中断一侧的神经传导束，以及术后护理的详细过程。...

本视频演示了一种利用高功率激光在黑腹果蝇（Drosophila）幼虫中诱导靶向神经元损伤的方法。将幼虫置于显微镜下定位后，先使用低功率激光扫描荧光标记的神经元，随后提高激光功率以诱导局部损伤。通过观察损伤部位荧光增强及形成凹坑来确认损伤效果。...

该视频展示了使用高冲击创伤（HIT）装置在Drosophila中诱导闭合性颅脑损伤（TBI）的方法。TBI引起机械性脑损伤，影响神经元功能，并随时间推移导致神经变性和死亡。...

本视频展示了在果蝇大脑中诱导穿透性创伤性脑损伤的方法。选取新羽化的果蝇，进行麻醉后置于配备荧光装置的体视显微镜下定位。识别表达GFP的蘑菇体，并插入微型针以造成损伤。随后让果蝇恢复，以供后续研究使用。...

来源：Shandra, O. 等，在重复性弥散性创伤性脑损伤小鼠模型中诱导创伤后癫痫。J. Vis. Exp.

本视频演示了使用侧向液体脉冲损伤（FPI）装置在大鼠中诱导创伤性脑损伤（TBI）的方法。该操作包括在麻醉大鼠的颅骨上钻孔以暴露硬脑膜，并将套管固定于其上。随后将大鼠连接至装置，装置产生的压力波传递至大脑，通过引起神经细胞死亡和血管损伤来模拟创伤性脑损伤。...

来源：Yang, Z., 等. 小鼠重复性脑震荡损伤模型. J. Vis. Exp.

来源：Logsdon, A. F., 等。用于啮齿类动物闭合性轻度创伤性脑损伤临床前评估的低强度冲击波模型。J. Vis. Exp.

来源：Macheda, T.，等。小鼠轻度创伤性脑损伤的电磁控制闭合式头部模型。J. Vis. Exp.

来源：Elzat, E. Y. Y.，等，基于微创技术建立小鼠挫伤性脊髓损伤模型。J. Vis.

来源：Lakshman, N., 等一种用于评估轻度脊髓损伤小鼠模型中内源性神经干细胞激活的神经球形成实验 视频实验杂志...

来源：Kumar, S.，等. 脊髓损伤后压疮的小鼠模型。J. Vis. Exp.

本视频演示了一种利用生物素标记法评估小鼠大脑皮层星形胶质细胞中靶蛋白内化的实验方法，随后进行细胞裂解、基于链霉亲和素的蛋白提取、变性处理及蛋白质印迹分析，以确认靶蛋白成功内化。...

本视频展示了一种利用海马神经元在图案化蛋白区域上评估蛋白活性的方法。神经元在测试蛋白区域表现出排斥现象，而在对照蛋白区域则表现出黏附现象，从而凸显了吸引性或排斥性蛋白活性的作用效果。...

本视频演示了通过向麻醉大鼠的巩膜上静脉注射高渗盐水以诱导青光眼的过程。盐水进入小梁网 通过...

使用匀浆器匀浆组织，以释放蛋白质，包括对脑功能至关重要的谷氨酸脱羧酶（GAD酶同工型）。...

本视频演示了在麻醉动物中通过手术暴露视神经并精确递送溶血卵磷脂以诱导局灶性脱髓鞘的过程，从而为后续研究建立神经病理学条件。...

将切片在含有荧光假性神经递质（FFNs）的 aCSF...

本视频展示了在感染赛勒病毒（Theiler's Murine Encephalomyelitis Virus,...

本视频展示了通过在受控环境中逐步升高体温，以在携带癫痫相关基因突变的转基因小鼠中诱导热诱导性癫痫发作的操作步骤。该方案用于研究癫痫的发作机制及潜在治疗方法。...

本视频展示了在表达光敏阳离子通道的转基因小鼠的冠状皮层脑片中诱发痫性事件的方法。将脑片置于记录室中，并将记录电极插入皮层浅层。当灌流阻断钾通道的致痫药物后，神经元变得过度兴奋，并产生自发性痫性事件。为了按需诱发痫性事件，可通过短暂的光脉冲激活锥体神经元中的阳离子通道，从而触发兴奋性神经递质的释放，继而诱导突触后神经元在短时间内产生痫性事件。...

本视频演示了使用戊四氮（PTZ）注射诱导小鼠癫痫的过程，PTZ 是一种 GABA-A 受体拮抗剂。通过反复腹腔注射...

来源：Soukupová, M.，等．在自由活动大鼠脑电图记录期间兴奋性氨基酸的微透析．J. Vis. Exp.

来源：Schönherr, S., 等. 结合光遗传学与冰冻断裂复型免疫标记技术研究小鼠杏仁核中谷氨酸受体的输入特异性分布。 J. Vis. Exp.

Source: Coniglio, S., et al. Coculture assays to study macrophage and microglia stimulation of glioblastoma invasion. J. Vis. Exp. (2016)
In this ...

Source: Mombo, B. N., et al. MitoCeption: transferring isolated human MSC mitochondria to glioblastoma stem cells. J. Vis. Exp. (2017)
In this video, ...

Source: Guyon, J., et al. A 3D Spheroid Model for Glioblastoma. J. Vis. Exp. (2020)The video demonstrates the generation of a three-dimensional ...

Source: Lim, M., et al. Modeling Brain Metastasis by Internal Carotid Artery Injection of Cancer Cells. J. Vis. Exp. (2022)In this video, an anesthetized ...

Source: Ear, P. H., et. al. Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids. J. Vis. Exp. (2019)This video demonstrates ...

Source: Mudduluru, G., et. al. Impedance-based Real-time Measurement of Cancer Cell Migration and Invasion. J. Vis. Exp. (2020)This video demonstrates an ...

Source: Bolcaen, J., et. al., PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator. J. Vis. Exp. (2017)This video ...

Source: Mendez, F. M. et al., Native Chromatin Immunoprecipitation Using Murine Brain Tumor Neurospheres. J. Vis. Exp.  (2018).This video ...

Source: Núñez, F. J., et al. Evaluation of Biomarkers in Glioma by Immunohistochemistry on Paraffin-Embedded 3D Glioma Neurosphere ...