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. 2019 Feb 20;9:2405. doi: 10.1038/s41598-019-39555-x

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© The Author(s) 2019

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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Involvement of convallatoxin in morphine antinociceptive tolerance and dependence. (A) Experiment flowchart for testing of drug effects on morphine tolerance and dependence. (B,C) Acute (B) and chronic (C) antinociceptive effects of each treatment in mice measured using tail-flick test. (B; upper panel) Treatment F_3,16 = 1938, min F_3,48 = 3828, interaction F_9,48 = 1089; (C; upper panel) Treatment F_3,16 = 80.31, min F_3,48 = 41.5, interaction F_9,48 = 18.41; all p < 0.001 (2-way ANOVA). Quantitative results from time-response curves are presented as AUC. (B; lower panel) F_3,16 = 1068; (C; lower panel) F_3,16 = 62.45; all p < 0.001 (1-way ANOVA). (D) Chronic convallatoxin diminished naloxone-precipitated withdrawal jumping. F_3,16 = 44.58, p < 0.001 (1-way ANOVA). (E) The acute antinociceptive effects of each treatment in morphine-tolerant mice. Treatment F_4,20 = 30.24, min F_3,60 = 81.9, interaction F_12,60 = 18.07; all p < 0.001 (2-way ANOVA; upper panel). Quantitative results from time-response curves, presented as AUC. F_4,20 = 26.54, p < 0.001 (1-way ANOVA; lower panel). (F) Experiment flowchart determining the acute drug effects on naloxone-precipitated withdrawal jumping in morphine-tolerant mice (upper panel). Acute convallatoxin diminished morphine withdrawal. F_3,16 = 38.56, p < 0.001 (1-way ANOVA; lower panel). (G) Representative immunofluorescence images of MOR (red) and WGA (green) distribution in the mouse DRG after chronic drug treatment. DAPI (blue) was a nuclear marker. Scale bar, 20 μm. Data in B (upper panel), C (upper panel), E (upper panel), **p < 0.01, ***p < 0.001 versus vehicle group. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 versus morphine-alone group (Bonferroni’s post hoc tests). Data in B (lower paner), C (lower panel), (D,E) (lower paner), (F), *p < 0.05, ***p < 0.001 versus vehicle group. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 versus morphine-alone group (Newman-Keuls post hoc tests). The values indicate the mean ± s.e.m. MPE, maximum possible effect.