A novel treadmill-based method for assessing learning and memory in mice

doi:10.12307/2026.717

Abstract

Abstract: BACKGROUND: Currently, behavioral tests for assessing learning and memory are important technical methods for the drug development and exploration of the pathogenesis of Alzheimer's disease. However, animal behavior is easily influenced by various factors, including external conditions, the function of internal organs and cells, and psychological states. Therefore, developing appropriate experimental methods that can effectively reflect animal cognitive behavior is an important prerequisite for exploring the pathogenic mechanism and prevention and treatment of Alzheimer’s disease.
OBJECTIVE: To verify that the treadmill test is a new method using two different mouse models and to explore the advantages and disadvantages of the treadmill test in evaluating the learning and memory ability of mice in comparison with the Morris water maze.
METHODS: The genotypes of APP/PS1 transgenic mice (Alzheimer’s disease group, n=8) and littermate wild-type mice (control group, n=8) and the pathological features of β-amyloid in the brain were identified using polymerase chain reaction and thioflavin S staining. Subsequently, 11-month-old APP/PS1 transgenic mice (Alzheimer’s disease group, n=8) and their littermates of wild-type mice (control group, n=8) were subjected to a treadmill test. At the same time, 8-month-old 3×Tg-AD mice (Alzheimer’s disease group, n=8) and wild-type control mice (control group, n=8) were subjected to treadmill and water maze tests sequentially.
RESULTS AND CONCLUSION: (1) The polymerase chain reaction results showed that the expression of APP and PSEN1 genes in APP/PS1 transgenic mice was higher than that in the control group (P < 0.001), and thioflavin S staining showed that there were more β-amyloid plaques in the hippocampus of APP/PS1 transgenic mice than in the control group (P < 0.001). The results of the treadmill test showed that the total number and duration of electric shocks in APP/PS1 transgenic mice were higher than those in the control group (P < 0.05). (2) The results of the treadmill test showed that the total number and duration of electric shocks in 3×Tg AD mice were higher than those in the control group (P < 0.05). The results of the water maze test showed that the escape latency of 3×Tg AD mice was longer than that of the control group from the 4th day of the localization navigation phase (P < 0.05). To conclude, the treadmill test can effectively reflect the learning and memory function of mice, and may become an experimental method for measuring mouse cognitive behaviors.

Key words: treadmill, learning and memory, APP/PS1 transgenic mice, 3×Tg-AD transgenic mice, behavior, Alzheimer’s disease

CLC Number:

Pei Xiaxia, Li Tian, Zhang Yanli, Gao Yanping, Su Qiang. A novel treadmill-based method for assessing learning and memory in mice[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(18): 4694-4701.

Figures/Tables 4

2.1 实验动物数量分析 APP/PS1转基因小鼠(n=8)及同窝对照野生型小鼠(n=8)、3×Tg-AD小鼠(n=8)及同窝对照野生型小鼠(n=8)均进入结果分析。 2.2 检测APP/PS1转基因及同窝对照小鼠的基因型及β-淀粉样蛋白病理改变通过聚合酶链式反应和硫磺素S染色确定APP/PS1转基因小鼠的基因型及脑病理特征是否发生。聚合酶链式反应结果显示，与对照组相比，APP/PS1转基因组小鼠的APP和PSEN1基因表达升高(P < 0.001)，见图1。硫磺素S可以结合阿尔茨海默病脑内β-淀粉样蛋白沉积物，是观察β-淀粉样蛋白斑块沉积的重要手段。硫磺素S染色结果显示，与对照组小鼠相比，APP/PS1转基因组小鼠脑内产生大量的β-淀粉样蛋白斑块沉积(P < 0.01)，见图2。 2.3 利用跑台实验检测APP/PS1转基因及同窝对照小鼠的学习记忆能力跑台实验测试中，APP/PS1转基因小鼠及野生对照小鼠总电击次数及总电击时间随跑步时间的变化曲线，如图3所示。在2 d的实验中，APP/PS1转基因组小鼠总电击次数及总电击时间均高于对照组，在第1天评估学习能力的实验中，两组小鼠总电击时间及总电击次数分别从跑步的第30分钟及第15分钟开始出现显著差异(P < 0.05)；在第2天评估记忆能力的实验中，两组小鼠总电击次数及总电击时间分别在第25分钟及第40分钟出现差异(P < 0.05)。 2.4 利用跑台实验检测3×Tg-AD小鼠及同窝对照小鼠的学习记忆功能 3×Tg-AD小鼠及同窝对照小鼠总电击次数及总电击时间随跑步时间的变化曲线，如图4所示。在2 d实验中，3×Tg-AD组小鼠总电击次数及总电击时间均高于对照组，第1天，3×Tg-AD 组小鼠总电击时间从第5分钟开始就高于对照组(P < 0.05)，总电击次数从第10分钟开始就高于对照组(P < 0.05)；第2天，3×Tg-AD组小鼠第1-20分钟总电击时间高于对照组(P < 0.05)，总电击次数没有出现有统计学意义的差异，这可能与使用的3×Tg-AD月龄小有关，但3×Tg-AD小鼠总电击次数仍出现了升高趋势，与上述APP/PS1转基因小鼠结果保持一致，表明6月龄3×Tg-AD小鼠相较于正常野生对照小鼠在跑台实验中出现了学习和记忆能力下降。 2.5 利用Morris水迷宫实验检测3×Tg-AD及同窝对照小鼠的学习记忆功能 3×Tg-AD及同窝对照小鼠的Morris水迷宫实验测试结果，见图5。从定位航行阶段的第4天开始，与野生对照小鼠相比，3×Tg-AD组小鼠逃避潜伏期长于对照组(P < 0.05)，见图5A，提示3×Tg-AD小鼠学习能力的确发生显著下降；在空间探索阶段，两组小鼠在目标象限停留时间和穿台次数比较差异无显著性意义(P > 0.05)，见图5B，C，但与对照组相比，3×Tg-AD组小鼠在目标象限停留时间和穿台次数有降低趋势，显示3×Tg-AD小鼠记忆能力确有受损，这与图4跑台为基础评估小鼠学习记忆能力的行为学实验所得出的结论相一致。图5D展示的是空间探索阶段两组小鼠的代表性轨迹图。"

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