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Journal of Peking University (Health Sciences) ›› 2023, Vol. 55 ›› Issue (2): 315-323. doi: 10.19723/j.issn.1671-167X.2023.02.016

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Establishment and behavioral evaluation of a mouse model of long-term free-choice alcohol drinking

Ting-ting YUAN1,Shen LI2,Yan WU2,Hai-tao WU1,2,*()   

  1. 1. Graduate Collaborative Training Base of Academy of Military Sciences, Hengyang Medical school, University of South China, Hengyang 421001, Hunan, China
    2. Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China
  • Received:2022-02-21 Online:2023-04-18 Published:2023-04-12
  • Contact: Hai-tao WU E-mail:wuht@bmi.ac.cn

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Abstract:

Objective: To establish a model of long-term free drinking mouse by feeding mice with alcohol to simulate the state of human voluntary long-term drinking, and on this basis, to further discuss the evaluation criteria of long-term free drinking mice model in sports, anxiety and cognitive behavior. Methods: Forty six-week-old SPF C57BL/6 male mouse were randomly divided into two groups: Long-term free drinking group (n=20) and normal control group (n=20). The two groups were given solid feed normally. The long-term free drinking group was free to take 10% alcohol and water every day, while the normal drinking group only took water every day. The mice were fed for 7 months, and were evaluated by a series of behavioral methods, including Rota-rod test, balance beam test, open filed test, the elevated plus maze, two-box social behavior, new object recognition, Y maze and water maze. Results: With the increase of drinking days, the mice showed significant alcohol addiction in the alcohol preference test. With the increase of alcohol intake, the mice in the long-term free choice drinking group had slightly shiny fur and reduced diet. Compared with the control group, the weight gain began to slow down from the third month, and the weight decreased significantly by the sixth and seventh months (P=0.006, P < 0.001). The mice showed reduced balance locomotion ability (P=0.003, P=0.001) in the rotary bar and balance beam test. In the open field and elevated cross test, the mice had obvious anxiety-like behavior (P < 0.001). The mice showed decreased social ability in the two boxes of social behavior (P < 0.016). In the experiment of new object recognition and Y maze, the exploration of new object decreased (P=0.018, P=0.040). In the water maze, cognitive functions, such as learning and spatial memory were reduced (P < 0.001). Conclusion: The successful establishment of the long-term free drinking mouse model is more convenient for us to carry out further research on the neural mechanism of alcohol addiction, and lays an experimental foundation for exploring the neural mechanism of alcohol addiction and related new targets.

Key words: Long-term, Free drinking, Mice model, Animal behavior

CLC Number: 

  • R749.62

Figure 1

Establishment of a long-term free drinking mice model and analysis of alcohol preference A, long-term free drinking group, n=20; B, normal control group, n=20; A, B, experimental procedure of modeling; C, most mice had an alcohol bias higher than 50% (n=18); D, effect of alcohol on the weight (#P < 0.01, ☆P < 0.001)."

Figure 2

Evaluation of exercise ability in rotarod test and balance beam in long-term free drinking mice A, C, schematic diagram of experimental device; B, D, long-term free drinking induced decreased motor balance ability of mice. Con, normal control group; Alc, long-term free drinking group. ns, the difference is not significant. *P < 0.05, # P < 0.01, ☆P < 0.001."

Figure 3

Evaluation of the anxiety-like behavior in long-term free drinking mice A, F, schematic diagram of experimental device; B, G, trajectory diagram (left) and heat map (right) of normal control group (top) and long-term free drinking group (bottom); C, D, E, long-term free drinking reduced the activity time, distance and total activity distance of mice in the central area of the open field; H, I, J, long-term free drinking reduced the open arm movement time, entry times and distance of the mice in the elevated cross. Con, normal control group; Alc, long-term free drinking group. ns, the difference is not signficant. ☆P < 0.001."

Figure 4

Social behavior evaluation of long-term free drinking mice A, schematic diagram of experimental device; B, heat maps of the normal control group (top) and the long-term free drinking group (bottom); C, both groups showed obvious sociability at sociability stage; D, there were significant social novelty in both groups, and the mice in the long-term free drinking group spent less time exploring stranger 2. Con, normal control group; Alc, long-term free drinking group. ns, the difference is not significant. ☆P < 0.001."

Figure 5

Evaluation of exploration of new objects in long-term free drinking mice A, E, schematic diagram of experimental device; B, F, heat maps of the normal control group (top) and the long-term free drinking group (bottom); C, there was no significant difference in the time consumed in the same object stage between the two groups (P=0.999, P=0.971); D, the normal control group showed significant exploration of new objects (P=0.003), while the long-term drinking group showed no significant difference in exploration of new objects (P=0.596), and there was also a difference in exploration of new objects between the two groups of mice (P=0.018); G, the time of the two groups of mice to explore the new arm (P=0.039); H, times of entering the new arm of mice in the two groups (P=0.580). Con, normal control group; Alc, long-term free drinking group. ns, the difference is not significant. *P < 0.05, ☆P < 0.001."

Figure 6

Behavioral evaluation of learning and memory ability in long-term free drinking mice A, schematic diagram of experimental device; B, C, trajectory diagram of the normal control group (top) and the long-term free drinking group (bottom); D, the time (latency) between entering the water and finding the platform of mice in the two groups; E, the distance between the two groups of mice entering the water and finding the platform; F, the speed of the two groups of mice from entering water to finding the platform; G, the number of times that mice in the two groups crossed the original platform area after removing the platform in the pool; H, the percentage of the time that the two groups of mice stayed in the target quadrant to the total time in this test; I, the percentage of the distance of the two groups of mice shuttling in the target quadrant to the total distance in the whole experiment. Con, normal control group; Alc, long-term free drinking group. ☆P < 0.001."

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