APPswe／PS1dE9／TAU三转基因阿尔兹海默病大鼠模型的建立

目的大鼠的大脑比小鼠更大，是研究神经系统的重要模型。建立APPswe／PS1dE9／TAU三转基因大鼠，发展能更全面表现人类阿尔兹海默病表型的动物模型。方法构建人PrP—hAPP695K595N／M596L、PrP-hPS1dE9和PDGF-TAU转基因表达载体，显微注射法制备转基因大鼠。PCR法鉴定转基因首建鼠及其子代基因型。Western blot检测转基因大鼠脑组织中人APP、PS1和TAU蛋白的表达。Morris水迷宫检测6月龄三转基因大鼠学习记忆能力改变。APP、PHF—TAU免疫组织化学染色观察三转基因大鼠脑组织APP及TAU的表达。结果得到1个同时高表达人APP、PS1和TAU三个基因的转基因大鼠品系。转基因大鼠6月龄已经出现显著的行为学改变：学习记忆能力下降，病理学改变表现为过度磷酸化TAU增多和神经元胞浆内AB表达异常增加。结论成功建立了APPswe／PS1dE9／TAU三转AD大鼠，可做为新一代工具动物模型用于基础医学和AD转化医学研究。

Establishment of APPswe/PS1dE9/TAU triple transgenic rat model of alzheimer disease

Objective To develop a model that could roundly show the phenotypes of human alzheimer disease （AD）, the triple-transgenie rat model harboring APP（ Swe）, PS1 dE9, and TAU transgenes was established in view of the advantage of rat as an important animal model on the research of nerve system. Methods APPswe/PS1dE9/TAU triple transgenic rat AD rats were generated on a SD background by co-injecting rat pronuclei with two human genes driven by the mouse prion promoter： ＇ Swedish＇ mutant human APP （APPsw） and exon 9 mutant human presenilin-1 （PSldEg） and human microtubule-associated protein tau gene under the control of PDGF promoter. Transgene integration was confirmed by genotyping and expression levels were evaluated by western blot （WB） of brain homogenates. The pathological changes were detected by human Abeta, TAU and Phospho-PHF-TAU immunohistoehemistry staining （IHC）. The behavioral and cognitive changes were evaluated by Morris water maze. Results One transgenie rat lines with high human APP （Swe）, PS1dE9, and TAU transgenic expression was selected from three transgenic founders. Compared with the wild type rat, the transgenic rat showed significant learning and memory impairments in the Morris water maze at 6 months of age. The triple transgenic rat manifested hyperphosphorylated tau and obvious aggregation of amyloid-β （Aβ） in the brain cortex and hippocampus. Conclusion APPswe/PS1dE9/TAU triple transgenic rat AD model was established. The triple transgenic AD rat fills a critical need for a next-generation animal model to enable basic and translational AD research.