Synphilin-1转基因小鼠模型的制备及帕金森病相关蛋白的表达  

Preparation of Synphilin-1 transgenic mouse model and study on the expression of Parkinson's disease-related protein

作　　者：金洪国[1] 唐维国[1] 金美玉[1] 何松彬[1] 周敏[1] 寇雪莲[1] 

机构地区：[1]温州医科大学附属舟山医院神经内科,浙江舟山316000

出　　处：《温州医科大学学报》2015年第9期660-664,670,共6页Journal of Wenzhou Medical University

基　　金：浙江省科技厅钱江人才计划项目(2010R10078)

摘　　要：目的:探讨Synphilin-1与帕金森病发病机制的相关性。方法:利用朊蛋白(Prp)启动子载体将人类Synphilin-1基因结合注入到小鼠受精卵中制作转基因(Tg)小鼠。通过小鼠尾活组织DNA的PCR检测确定小鼠基因型。然后进行PD相关蛋白免疫印迹分析,脑组织切片免疫组织化学分析,双标记免疫荧光染色及Tg小鼠表型与运动能力分析。结果:Synphilin-1广泛表达于包括黑质的Tg小鼠大脑的神经元中,但没有观察到多巴胺(DA)神经元的大量流失,Synphilin-1蛋白在Tg小鼠的大脑中被多泛素化,部分呈现不溶性。利用改良SHIRPA法观察Tg组小鼠与非Tg组小鼠在行为和形态学上的改变,差异无统计学意义。通过加速转圈测试和足迹分析观察2组小鼠的运动能力,Tg组小鼠与非Tg组小鼠相比运动能力减少。结论:Synphilin-1可能参与运动功能的调节,但不影响脑组织内PD相关蛋白的表达,对DA神经元亦无显著影响。Objective：To explore the pathogenesis correlafion of Synphilin-1 with Parkinson＇s disease.Methods：Generated transgenic mice expressing human Synphilin-1 under the prion protein prom-oter.The genotype of offspring mice was determined by a PCR assay of DNA obtained from tail biopsies.The human Synphilin-1 gene was injected into the fertilized ovum using prion promoter vector to generate the transgenic mice,then PD-related protein immunoblot analysis,the brain tissue slice analysis with immunohistochemistry,double labeling imtnunofluorescence staining and the phenotypic and athletic analysis of transgenic mice were performed.Results：Synphilin-1 was widely expressed in neurons in the brain,including the substantianigra,but massive loss of dopamine neurons was not observed.In the transgenic mouse brain,Synphilin-1 protein was polyubiquitinated and partially insoluble.Although modified-SHIRPA revealed no significant difference in behavior and morphology,the reduced rotarod performance and step length were observed in transgenic mice as compared with non-transgenic littermates.Conclusion：Synphilin-1 may be involved in motor function,and its accumulation in the central nervous system can cause motor impairments.Synphilin-1 may be involved in regulation of motor function,but had no significant effect of the expression of Parkinson＇s disease related proteins nor dopamine neurons in the brain tissue.

关 键 词：SYNPHILIN-1 帕金森病 转基因小鼠 抗酪氨酸羟化酶 Α-共核蛋白 

分 类 号：R742.5[医药卫生—神经病学与精神病学]