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核定位人源α-突触核蛋白转基因小鼠的建立

Establishment of the human α-synuclein nuclear localization signal transgenic mice

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
目的 建立一种核定位人源α-突触核蛋白(α-synuclein,α-syn)转基因小鼠,并探究人源α-syn核输入对小鼠行为的影响.方法 构建h SNCA-NLS和EGFP的慢病毒载体,利用显微注射法建立转基因小鼠模型,通过PCR和Western Blot方法鉴定转基因首建鼠及其子代基因型和蛋白表达情况.采用免疫荧光鉴定人源α-syn在小鼠脑组织中的共定位情况,同时采用旷场、转棒、O迷宫实验评价转基因小鼠的行为改变情况.结果 h SNCA-NLS基因成功插入小鼠基因组中,人源α-syn成功表达,并且有明显的核定位现象.进一步研究发现,核定位人源α-syn转基因小鼠在 2 月龄时开始表现出了运动功能障碍,发生了星形胶质细胞增生和炎症反应,于 9 月龄时表现出了明显的焦虑样症状,γ-氨基丁酸(GABA)基因表达减少,这些表现一直持续到小鼠12 月龄.结论 成功构建了核定位人源α-syn转基因小鼠,小鼠表现出明显的运动功能障碍和焦虑样症状.该模型的成功建立可以为研究α-syn核定位现象在帕金森病中的作用提供一定的研究基础.
Objective To establish a human α-synuclein nuclear localization signal transgenic mouse model and investigate the effects of α-synuclein nuclear localization on the behavior of mice.Methods Human α-synuclein nuclear localization signal and EGFP lentiviral vectors were constructed.Transgenic mice were created with the microinjection method.Using PCR and Western Blot method to identify the genotypes and protein expression of the transgenic founder mice and their offsprings.The immunofluorescence was used to examine the localization of human α-synuclein in the mouse brain tissue.The behavioral changes of the transgenic mice were evaluated by the open field test,rotarod test,and O maze test.Results The h SNCA-NLS gene was successfully inserted into the mouse genome,the human α-syn was successfully expressed,and the human α-syn has localized with the nuclear.Further studies found that human α-synuclein nuclear localization signal transgenic mice had significant motor dysfunction,astrocyte proliferation and inflammatory response at 2 months of age and exhibited significant anxiety-like symptoms and reduced expression of the γ-aminobutyric acid(GABA)gene at 9 months of age,which persisted until 12 months of age.Conclusions A human α-synuclein nuclear localization signal transgenic mouse model has been successfully established.The mice exhibit significant motor dysfunction and anxiety-like symptoms.The successful establishment of this model provides a foundation for studying the role of α-syn nuclear localization in Parkinson's disease.

human α-synucleinmousenuclear localizationtransgenicmotor dysfunction

韦梦晨、范胜涛、吴海婷、张艺薇、王子鸥、黄璋琼

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北京协和医学院 中国医学科学院医学生物学研究所,昆明 650118

人源α-突触核蛋白 小鼠 核定位 转基因 运动功能障碍

云南省技术创新人才培养对象项目云南省技术创新人才培养对象项目中国医科院医学与健康科技创新工程项目国家自然科学基金联合基金科技创新2030-重大专项

202105AD160018202005AD1600062021-I2M-1-043U22022142023ZD0406306

2024

10.3969/j.issn.1005-4847.2024.03.004

中国实验动物学报
中国实验动物学会，中国医学科学院医学实验动物研究所

中国实验动物学报

CSTPCD北大核心
影响因子：0.767
ISSN：1005-4847
年,卷(期)：2024.32(3)