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. 2024 Sep 12;21(5):168.
doi: 10.3892/br.2024.1856. eCollection 2024 Nov.

Behavioral video coding analysis of chronic morphine administration in rats

Jie Yin  1 , Tiecheng Zhang  2 , Dan Li  3 , Fan Xu  2 , Huan Li  2 , Xinyu Pan  2 , Fang Liu  2 , Yongqi Zhao  1 , Xiechuan Weng  1
Affiliations

Behavioral video coding analysis of chronic morphine administration in rats

Jie Yin et al. Biomed Rep. .

Abstract

The present study assessed the behavior of morphine-addicted rats using behavioral video coding technology, to evaluate effective methods for identifying morphine addiction. Rats were divided into a control group (n=15) and a morphine addiction group (n=15). The morphine addiction model was established with a 14-day increasing dose scheme, confirmed using a conditional place preference (CPP) experiment. After successful modeling, the rats' behavior was recorded for 12 h, then coded and analyzed using Observer XT behavior analysis software. Compared with the control group, morphine-addicted rats showed increased heat pain tolerance time (P=0.039) and spent more time in the white box during the CPP experiment (P<0.001). Video coding analysis revealed significant behavioral changes in morphine-addicted rats compared to controls. In addition to being lighter, morphine-addicted rats showed decreased water intake, reduced licking of forelimbs and hind limbs, and altered sleeping posture (sleeping curled up) during the day (all P<0.05). In conclusion, chronic morphine administration in rats leads to distinctive behavioral changes, including decreased licking frequency, reduced water intake and altered sleep posture. Video coding analysis, as a safe and non-invasive method, may provide a convenient and efficient approach for studying morphine addiction in rats.

Keywords: addiction; conditional place preference experiment; morphine; rat; video coding.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Body weight changes during morphine addiction modeling in rats. Body weight changes over the 14-day modeling period for rats in the morphine and control groups. On day 14, the average body weight of control rats was 233.6±8.0 g (maximum, 287.7 g), while that of rats in the morphine group was 206.2±3.4 g (maximum, 222.0 g). There was a statistically significant difference in the rate of body weight gain for rats in the morphine group compared with the control group (n=15, day 14, z=2.717, P=0.0057). *P<0.05.
Figure 2
Figure 2
Changes in thermal nociceptive tolerance time during morphine addiction modeling in rats. Thermal nociceptive tolerance time over 14 days for control rats and rats in the morphine group. The gap in thermal nociceptive tolerance time before and after morphine injection was more pronounced in the morphine group compared with the control group before and after saline injection (n=15, U=74.4, P=0.039). *P<0.05.
Figure 3
Figure 3
Comparison of white box time in CPP experiment before and after morphine addiction modeling. Comparative analysis of the time spent in the white box during the CPP experiment between rats in the morphine group and the control group. The comparisons include differences in white box time before morphine addiction (day 4) and after morphine addiction (day 14), and the variation in white box time for rats in the morphine group before and after addiction. Prior to morphine addiction (day 4), no significant difference was observed in the white box time between rats in the morphine group and the control group (n=10, χ²=0.2, P=0.64). However, after morphine addiction (day 14), a significant difference in white box time was evident between the two groups (n=10, χ²=134.8, P<0.001). Additionally, a significant difference in white box time was identified in the morphine group between day 4 and day 14 (n=10, χ²=170.8, P<0.001). ***P<0.001. CPP, conditioned place preference; ns, not significant.
Figure 4
Figure 4
Behavioral analysis by video coding comparing rats in the morphine group and control group. (A) No significant difference was shown in feed intake between the morphine and control groups (n=8, U=65.5, P=0.98). (B) A significant difference was shown in water intake, with the morphine group drinking less than the control group (n=8, U=17, P=0.0014). (C) A significant difference was shown in foreleg licking, with the morphine group licking less than the control group (n=8, U=27, P=0.014). (D) A significant difference was shown in hind limb licking, with the morphine group licking more than the control group (n=8, U=32, P=0.035). (E) No significant difference was shown in hind leg lifting between the morphine and control groups (n=8, U=62, P=0.082>0.05). *P<0.05.
Figure 5
Figure 5
Behavioral comparison between morphine and control group rats over a 9-h period using video coding. (A) Comparison of ‘ball sleeping’ behavior, a significant difference was observed only during the 2:30-3:00 p.m. period (n=8, z=-2.412, P=0.0159). (B) Analysis of ‘marten sleeping behavior (n=8, P=0.8>0.05). (C) Frequency of scratching actions, a significant difference was found only during the 1:00-1:30 p.m. period (n=8, z=-1.964, P=0.0495). (D) Frequency of fur licking, a significant difference was observed only during the 3:30-4:00 p.m. period (n=8, z=-2.29, P=0.022). (E) Comparison of walking frequency, significant differences were noted during the 12:00-12:30 p.m. (n=8, z=-2.093, P=0.036) and 6:00-6:30 p.m. periods (n=8, z=2.039, P=0.0415). *P<0.05.
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Grants and funding

Funding: This study was supported by the National Natural Science Foundation of China (grant no. 82073833) and the National Key R&D Plan (grant nos. 2022YFC3600500 and 2022YFC3600502).

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