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. 2023 Sep 18;8:355. doi: 10.1038/s41392-023-01578-2

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Rox promotes Mor-CPP extinction and inhibits Mor-CPP reinstatement. a The paradigm of Mor-CPP formation, extinction and reinstatement. As described in Methods section (Pattern 1), the CPP established mouse was trained to extinct, Rox was administrated instate of morphine after postconditioning phase for 3 cycles and a single Rox injection was conducted 6 h before reinstatement test. b Time spent in drug-paired box in Mor-CPP formation and extinction periods. Rox (10 mg/kg) promoted extinction of Mor-CPP as indicated by faster reduction in CPP scores (Veh, Mor and Rox, n = 10; group, F_(2,135) = 24.26, P < 0.0001; period, F_(4,135) = 4.217, P = 0.0030; group × period, F_(8,135) = 3.894, P = 0.0004). c Saline injection did not induce reinstatement of Mor-CPP (Veh, Mor and Rox, n = 10; F_(2,27) = 0.1959, P = 0.8232). d Mor (5 mg/kg) triggered Mor-CPP reinstatement, whereas it was inhibited by Rox (10 mg/kg) treatment (Veh, Mor and Rox, n = 10; F_(2,27) = 7.725, P = 0.0022). e The paradigm of Mor-CPP formation, extinction and reinstatement. As described in Methods section (Pattern 2), the mouse, CPP well established, was trained to extinct by repeat preference tests for 5 consecutive days, Rox was administrated instate of morphine after postconditioning phase for 3 cycles and a single Rox injection was conducted 6 h before reinstatement test. f Mor group was divided into Mor1 and Mor2 groups. Time spent in Mor-paired box in CPP formation and extinction periods (Veh, n = 12; Mor1, n = 14; Mor2, n = 12; group, F_(2,245) = 1.862, P = 0.1576; period, F_(6,245) = 2.431, P = 0.0266; group × period, F_(12,245) = 1.173, P = 0.3030). g Saline injection did not induce reinstatement of Mor-CPP (Veh, n = 8; Mor1, n = 9; Mor2, n = 8; group, F_(2,22) = 0.4210, P = 0.6616). h Mor (5 mg/kg) successfully triggered Mor-CPP reinstatement, whereas it was inhibited by Rox (10 mg/kg) treatment (Veh, Mor and Rox, n = 8; F_(2,21) = 7.110, P = 0.0044). Values are presented as means ± SEM. Statistical analyses for b, f and and for c, d, g and h were performed using two-way ANOVA and one-way ANOVA followed by Bonferroni-corrected tests, respectively. ^*P < 0.05, ^**P < 0.01; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 Mor or Mor1 vs Veh; ^{^^}P < 0.01, ^{^^^}P < 0.001 Rox or Mor2 vs. Veh; ^$P < 0.05 Mor vs Rox^; NS: not significant. Veh: vehicle; Sal: saline