Table 1.
Primer sequences used for semi-quantitative real time PCR gene expression study of corticotropin-releasing factor type 1 receptor, corticotropin-releasing factor type 2 receptor and prodynorphin.
Figure 1.
Conditioned Place Preference (CPP) test for morphine in adult zebrafish.
(A) Baseline preference of fish between dotted vs. white compartment as shown by percentage of initial time spend in compartment (n = 23). (B) Preference behavior responses in the CPP tanks for control group as shown by percentage of time spent in conditioning compartment before (white bar) and after (grey bar) conditioning. (n = 15). (C) Place preference behavior of adult zebrafish treated different morphine concentration (Control, n = 15; 0.5 mg/L, n = 18; 1.5 mg/L, n = 16; 3 mg/L, n = 15). Mean values were subjected to one way ANOVA and where applicable, ranked by Tukey test (P<0.05); mean percentage values with same letter are not significantly different.
Figure 2.
Effects of morphine treatment, morphine withdrawal and mitragynine on swimming behavior in adult zebrafish using the novel tank diving test.
(A). Effect of two weeks chronic morphine treatment (1.5 mg/L), 24 h morphine withdrawal and morphine withdrawal in presence of 2 mg/L mitragynine on swimming behavior of zebrafish. (Control, n = 17; morphine treatments, n = 25; morphine withdrawal, n = 22; morphine withdrawal+mitragynine, n = 22). All data were presented in mean ± SEM. Mean values were subjected to one way ANOVA and where applicable, ranked by Tukey test (P<0.05); mean values with same letter are not significantly different. (B) Effect of 20 min acute mitragynine treatment (1 mg/L and 2 mg/L) on swimming behavior of zebrafish. (Control, n = 21; 1 mg/L mitragynine, n = 20; 2 mg/L mitragynine, n = 21). All data were presented in mean ± SEM. Mean values were subjected to one way ANOVA and where applicable, ranked by Tukey test (P<0.05); mean values with same letter are not significantly different.
Figure 3.
Effect of two weeks chronic morphine treatment (1.5 mg/L), 24-h morphine withdrawal and morphine withdrawal in presence of mitragynine (1 mg/L or 2 mg/L) on whole-body cortisol (µg/g fish).
Data are presented as mean ± SEM. (Control, n = 16; morphine treatments, n = 14; morphine withdrawal, n = 21; 1.0 mg/L Mitragynine, n = 23; 2 mg/L Mitragynine, n = 20). Mean values were subjected to one way ANOVA and where applicable, ranked by Tukey test (P<0.05); values with same letter are not significantly different.
Figure 4.
Effect of two weeks chronic morphine treatment (1.5 mg/L), 24-h morphine withdrawal and morphine withdrawal in presence of 2 mg/L mitragynine on the mRNA expression of CRF-R1, CRF-R2 and PDYN in zebrafish.
Data are presented as mean ± SEM of three independent experiments. Mean values were subjected to one way ANOVA and where applicable, ranked by Tukey test (P<0.05); mean percentage values with same letter are not significantly different.