神经元与小胶质细胞示踪及可清除小鼠模型构建  

2023,20(1):41.Construction of a mouse model to trace neurons and microglia and ablate microglia for investigating their interactive roles

作　　者：陈梅 张准宏 楚世峰 张钊 陈乃宏 CHEN Mei;ZHANG Zhun-hong;CHU Shi-feng;ZHANG Zhao;CHEN Nai-hong(Institute of Traditional Chinese and Zhuang-Yao Ethnic,Guangxi University of Chinese Medicine,Nanning 530000,China;State Key Laboratory of Bioactive Substances and Functions of Natural Medicines,Institute of Materia Medica&Neuroscience Center,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]广西中医药大学中医药壮瑶医药研究院,广西南宁530000 [2]中国医学科学院药物研究所神经科学中心,天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《中国药理学通报》2025年第5期994-999,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82374060);中国医学科学院医学与健康科技创新工程项目(No 2021-I2M-1-020)。

摘　　要：目的构建一种小胶质细胞和神经元示踪小鼠模型用于实时分析小胶质细胞和神经元的动态变化,并通过小胶质细胞清除解析其与神经元互作的功能作用。方法小胶质细胞示踪鼠与神经元示踪鼠杂交构建Tmem119-e(2A-tdTomato-2A-DTR);Thy1-GFP双转鼠,提取鼠尾基因组检测双转基因型;应用共聚焦及双光子显微镜检测小鼠神经元形态;利用活体双光子成像技术观察脑部小胶质及神经元分布;利用白喉毒素腹腔注射检测小胶质细胞的清除效果。结果双转鼠基因型正确,小胶质细胞和神经元均可实现可视化,且小胶质细胞可被白喉毒素定向清除。结论成功构建神经元及小胶质细胞示踪小鼠模型,该模型将有助于活体实时观测生理或病理状态下小胶质与神经元的变化,明确其在脑稳态中的具体作用。Aim To generate a dual-reporter mouse model for tracing microglia and neurons to analyze their dynamic changes in real-time and to investigate the functional role of microglia-neuron interactions through microglial depletion.Methods Microglia reporter mice were crossed with neuron reporter mice to generate Tmem119-e(2A-tdTomato-2A-DTR);Thy1-GFP dual-reporter mice.Genotyping was performed using tail DNA.Confocal and two-photon microscopy were used to examine neuronal morphology.In vivo two-photon imaging was employed to observe the distribution of microglia and neurons in the brain.Diphtheria toxin(DT)was intraperitoneally injected to deplete microglia.Results The dual-reporter mice were successfully generated with the correct genotype.Both microglia and neurons were visualized,and microglia were specifically depleted by DT.Conclusions The successful establishment of a dual-reporter mouse model for tracing neurons and microglia will facilitate real-time in vivo observation of microglial and neuronal changes under physiological or pathological conditions,elucidating their specific roles in brain homeostasis.

关 键 词：小胶质细胞 神经元 脑稳态 活体可视化 双光子成像 中枢神经系统疾病 

分 类 号：R-332[医药卫生] R322.81R363-332R742