Abstract: Objective This study aims to provide reference and theoretical basis for selecting rTg4510 transgenic mice for the study of phosphorylated Tau protein metabolism, AD related pathogenic mechanisms, and drug screening. Method The phosphorylated tau protein, amyloid beta( Aβ) plaque and microglia in the brain of rTg4510 transgenic mice and AD patients were detected by immunofluorescence staining with antibodies ( S396, 4G8, IBA1 ) , software ( CaseViewer, IPWIN Application, and GraphPad Prism 8) were used to analyze the results, the expression, distribution and type of phosphorylated tau protein aggregation in the brain of rTg4510 transgenic mice and AD patients, as well as the characteristics of microglia lesions and Aβ plaques were compared. Result Similar to the characteristics that the expression of phosphorylated Tau protein ( S396 ) in cerebral cortex and hippocampus of AD patients was significantly higher than that of healthy control group ( p<0. 001) , the expression of phosphorylated Tau protein ( S396 ) in cerebral cortex and hippocampus of rTg4510 transgenic mice was significantly higher than that of wild-type control mice ( p< 0. 001) . Similar to the characteristics that the expression of phosphorylated Tau protein ( S396) in cerebral cortex of AD patients was significantly higher than that in hippocampus ( P < 0. 001) , the expression of phosphorylated Tau protein ( s396) in the cerebral cortex of rTg4510 transgenic mice has a similar trend compared with the expression of phosphorylated Tau protein ( s396) in the hippocampus. Similar to AD patients, type 1 and type 2 aggregation of hyperphosphorylated tau protein ( S396 ) were found in the brain of rTg4510 transgenic mice. Similar to AD patients, rTg4510 transgenic mice brain microglia ( Iba1) are sensitive to the recognition of hyperphosphorylated Tau protein ( S396) . Different from the abnormal expression of Aβ plaques (4G8) associated with hyperphosphorylated Tau protein ( S396) in the brain of AD patients, the abnormal expression of Aβ plaques ( 4G8 ) was not associated with hyperphosphorylated Tau protein ( S396) in the brain of rTg4510 transgenic mice. Conclusion rTg4510 transgenic mice will be a useful animal model for studying the pathogenic mechanism of AD related phosphorylated Tau protein and drug development.

Key words: Alzheimer’ s disease, Tau protein, microglia, Aβ plaque, rTg4510 transgenic mice

摘要： 目的 本研究拟为选择 rTg4510 转基因小鼠用于磷酸化 Tau 蛋白代谢、AD 相关致病机制及药物筛选等研究提供参考和理论依据。 方法 利用免疫荧光染色通过 S396、4G8、IBA1 等抗体检测了 rTg4510 转基因小鼠与 AD 患者脑内磷酸化 Tau 蛋白、β 淀粉样蛋白( Aβ) 斑块和小胶质细胞,利用 CaseViewer、IPWIN Application 和 GraphPadPrism 8 软件对三者进行了分析,比较了 rTg4510 转基因小鼠与 AD 患者脑内磷酸化 Tau 蛋白聚集的表达、分布、类型以及小胶质细胞病变、Aβ 斑块等特征的异同。 结果 与 AD 患者大脑皮质及海马中磷酸化 Tau 蛋白( S396) 的表达显著高于健康对照组的特征相似(P<0. 001) ,rTg4510 转基因小鼠大脑皮质及海马中磷酸化 Tau 蛋白( S396)的表达显著高于野生对照小鼠(P<0. 001) 。 与 AD 患者大脑皮质中磷酸化 Tau 蛋白( S396)表达显著高于相同面积内海马区磷酸化 Tau 蛋白( S396) 表达的特征相近( P < 0. 001) ,rTg4510 转基因小鼠大脑皮质中磷酸化 Tau 蛋白( S396)表达相对于相同面积内海马区磷酸化 Tau 蛋白( S396) 的表达也有类似趋势。 与 AD 患者相似,rTg4510 转基因小鼠脑内过度磷酸化 Tau 蛋白( S396)聚集均可见 type1 类型和 type2 类型。 与 AD 患者相似,rTg4510 转基因小鼠脑内小胶质 细 胞 ( IBA1) 对 过 度 磷 酸 化 Tau 蛋 白 ( S396) 识 别 敏 感。 与 AD 患 者 脑 内 过 度 磷 酸 化 Tau 蛋 白( S396)伴随出现 Aβ 斑块(4G8)异常表达不同的是,rTg4510 转基因小鼠脑内过度磷酸化 Tau 蛋白( S396)未伴随出现 Aβ 斑块(4G8)异常表达。 结论 rTg4510 转基因小鼠将是用于研究 AD 相关磷酸化 Tau 蛋白致病机制和药物研发等方面可用的动物模型。

关键词: 阿尔茨海默病, Tau 蛋白, 小胶质细胞, Aβ 斑块, rTg4510 转基因小鼠