APP/PS1+SCN2B-/-转基因小鼠模型的建立及鉴定  

Establishment and Identification of APP/PS1+SCN2B-/- Transgenic Mouse Model

作　　者：檀雅欣 李珊 王廷华 习杨彦彬 TAN Ya-xin;LI Shan;WANG Ting-hua;XI YANG Yan-bin(Institute of Neuroscience,Basic Medical College,Kunming Medical University,Kunming Yunnan 650500,China)

机构地区：[1]昆明医科大学基础医学院神经科学研究所

出　　处：《昆明医科大学学报》2019年第12期7-12,共6页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(81960210,81560238);昆明医科大学认知功能障碍相关脑疾病研究科技创新团队基金资助项目(CXTD201807);云南省应用基础研究计划基金资助项目(2016FB123)

摘　　要：目的建立APP/PS1+SCN2B-/-转基因小鼠,为深入研究SCN2B基因在AD发生发展中的作用提供实验工具。方法将APP/PS1转基因小鼠与SCN2B-/-转基因小鼠合笼,子代同胞兄妹交配,子代的后代中选取APP/PS1高表达和SCN2B阴性的小鼠再同胞兄妹交配,并培育。得到的目的基因的小鼠与WT小鼠对照,进行表型鉴定。运用PCR鉴定子代小鼠尾部组织基因组DNA,用RT-PCR验证Aβ和SCN2B在大脑中的蛋白的表达,通过免疫荧光观察SCN2B在转基因小鼠神经系统中的表达,最后用水迷宫在行为学上对转基因鼠进行鉴定。结果经PCR鉴定成功构建APP/PS1+SCN2B-/-转基因鼠杂交群体,并扩群培育,经免疫染色和RT-PCR分析转基因小鼠的Aβ和SCN2B在大脑中的蛋白的表达,发现Aβ在转基因鼠的海马和皮质中高表达,SCN2B在海马中几乎检测不到,免疫染色也未发现SCN2B在神经系统中的表达。水迷宫结果显示转基因鼠与同月龄野生型(WT)小鼠相比在空间记忆和认知差异有统计学意义(P<0.05)。结论成功构建APP/PS1+SCN2B-/-转基因小鼠模型,从基因型、组织学表现到行为学表现均具备了2种品系转基因鼠的特质,为进一步研究SCN2B基因在AD发生发展中的作用提供了动物研究模型。Objective To establish an APP/PS1+SCN2B-/-transgenic mouse model, so as to provide the experimental tool for studying the role of SCN2B in the pathogenesis of AD. Methods APP/PS1 AD model mice were crossed with SCN2 B-/-transgenic mice. Offspring, SCN2 B negative with high APP/PS1 expression, were selected,of females mated to full siblings. The genotype was determined by PCR. The expression of SCN2B and Aβin tissues was detected by immumohistochemical staining and RT-PCR. Finally, the transgenic mice’behavioral surveillance were identified by water maze. Results APP/PS1+SCN2B-/-transgenic mice were detected by PCR. The expression level of Aβ significantly increased in the prefrontal cortex and hippocampus and SCN2B was few in hippocampus of APP/PS1+SCN2B-/-Tgmice. As for immumohistochemical staining, there was no expression of SCN2B in the nervous system.Spatial learning and memory were decreased in APP/PS1/-SCN2B-/-Tg mice compared with wild-type( WT) mice. Conclusions APP/PS1+SCN2B-/-transgenic mouse model was successfully established. It can be used as animal model for further research of the role of SCN2B in the pathogenesis of AD.

关 键 词：电压门控性钠通道二型beta亚单位 APP/PS1 转基因小鼠 阿尔茨海默病 

分 类 号：R73-3[医药卫生—肿瘤]