Department of Forensic Medicine, Tokyo Medical University, Shinjyuku-ku, Tokyo, Japan.
PLoS One. 2020 Oct 8;15(10):e0236606. doi: 10.1371/journal.pone.0236606. eCollection 2020.
In this study, we investigated locomotor activity and responses to repeated light and dark stimuli to assess cannabinoid-induced abnormal behavior in zebrafish larvae (Danio rerio), as an alternative to standard rodent models. To induce the desired responses, we used cannabidiol and WIN55,212-2, two major cannabinoid components. A repeated light and dark test was used to assess how drug exposure influences locomotory responses. Larvae were examined after moderate cannabidiol and WIN55,212-2 exposure and at 24 h after transfer to untreated water. We found that cannabidiol did not produce a dose-dependent inhibitory effect on locomotor activity, with both 0.5 and 10 μg/mL concentrations reducing movement velocity and the total distance moved. However, 10 μg/mL cannabidiol was observed to attenuate the responses of larvae exposed to darkness. No differences were detected between the control and cannabidiol-treated groups after 24 h in fresh water. Fish treated with WIN55,212-2 at 0.5 and 1 μg/mL showed virtually no activity, even in darkness, whereas a concentration of 10 μg/mL induced mortality. A 24-h period in fresh water had the effect of reversing most of the drug-induced immobilization, even in the WIN55,212-2-treated groups. Larvae were also evaluated for their responses to cannabidiol subsequent to an initial exposure to WIN55,212-2, and it was accordingly found that treatment with cannabidiol could attenuate WIN55,212-2-induced abnormal immobilization, whereas equivalent doses of cannabidiol and WIN55,212-2 produced a mixed response. In conclusion, the behavioral effects of the two cannabinoids cannabidiol and WIN55,212-2 appear to be ratio dependent. Furthermore, the repeated light and dark test could serve as a suitable method for assaying drug-induced behavior.
在这项研究中,我们研究了斑马鱼幼虫(Danio rerio)的运动活性和对重复光照和黑暗刺激的反应,以评估大麻素诱导的异常行为,作为标准啮齿动物模型的替代方法。为了诱导所需的反应,我们使用了大麻二酚和 WIN55,212-2,这两种主要的大麻素成分。重复的光照和黑暗测试用于评估药物暴露如何影响运动反应。在中等剂量的大麻二酚和 WIN55,212-2 暴露后以及转移到未处理的水中 24 小时后,对幼虫进行了检查。我们发现,大麻二酚对运动活性没有产生剂量依赖性的抑制作用,0.5 和 10 μg/mL 浓度都降低了运动速度和移动的总距离。然而,10 μg/mL 大麻二酚被观察到减弱了暴露于黑暗的幼虫的反应。在新鲜水中 24 小时后,对照组和大麻二酚处理组之间没有发现差异。在 0.5 和 1 μg/mL 浓度下,WIN55,212-2 处理的鱼类几乎没有活动,即使在黑暗中也是如此,而 10 μg/mL 浓度则诱导了死亡。在新鲜水中 24 小时的处理效果是逆转了大部分药物诱导的固定,即使在 WIN55,212-2 处理组中也是如此。还评估了幼虫对大麻二酚的反应,随后对 WIN55,212-2 进行了初始暴露,结果发现,大麻二酚的处理可以减轻 WIN55,212-2 诱导的异常固定,而等效剂量的大麻二酚和 WIN55,212-2 则产生了混合反应。总之,两种大麻素大麻二酚和 WIN55,212-2 的行为效应似乎是比例依赖性的。此外,重复的光照和黑暗测试可以作为一种合适的方法来检测药物诱导的行为。