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目的利用斑马鱼这一新型模式动物建立行为模型,考察成瘾性药物甲基苯丙胺对于斑马鱼游泳行为的影响及谷氨酸转运体表达量的变化。方法利用斑马鱼行为视频跟踪分析系统自动记录动物行为参数,运动路程、穿越次数及在顶部停留时间作为测试指标,采用甲基苯丙胺5 mg·L-1,20 mg·L-1和50 mg·L-1,分别考察急性暴露及作用24 h后对于斑马鱼行为影响,之后利用实量定量PCR考察质膜型谷氨酸转运体(GLT1)和囊泡型谷氨酸转运体(VGLUT)表达量变化。结果急性暴露条件下,与正常组相比,斑马鱼垂直方向的穿越次数及在上半区停留时间呈非常显著的下降(P<0.01),且这种降低与甲基苯丙胺浓度有剂量依赖关系。在甲基苯丙胺急性处理后24 h,斑马鱼垂直方向的穿越次数及在上半区停留时间仍呈显著的下降(P<0.01)但不具有剂量依赖关系。急性暴露后斑马鱼脑GLT1显著下调(P<0.05),VGLUT3显著上调(P<0.01)。24 h后GLT1的表达量有所上调,VGLUT3上调趋势更加显著(P<0.01),但均无剂量关系。结论建立了甲基苯丙胺致斑马鱼神经损伤模型,甲基苯丙胺对于斑马鱼行为产生特征性影响且具有剂量关系。
OBJECTIVE: To establish a behavioral model using a novel model of zebrafish to investigate the effect of addictive methamphetamine on zebrafish swimming behavior and glutamate transporter expression. Methods The zebrafish behavioral video tracking system was used to automatically record the parameters of animal behavior, the distance traveled, the number of traversing times and the dwell time at the top as the test indexes. Methamphetamine 5 mg · L-1, 20 mg · L-1 and 50 mg · L-1, respectively. The effects on the behavior of zebrafish after acute exposure and for 24 h were investigated. Then the expression of GLT1 and VGLUT were examined by quantitative real-time PCR The amount of change. Results Under the conditions of acute exposure, the number of vertical crossing of the zebrafish and the retention time in the upper half of the zebrafish significantly decreased compared with the normal group (P <0.01), and this reduction was dose-dependent with the concentration of methamphetamine . At 24 h after acute methamphetamine treatment, the number of crossings in the vertical direction and the dwell time in the upper half of the zebrafish were still significantly decreased (P <0.01) but not in a dose-dependent manner. After acute exposure, GLT1 in brain of zebrafish significantly decreased (P <0.05) and VGLUT3 increased significantly (P <0.01). After 24 h, the expression of GLT1 was up-regulated, while the up-regulation of VGLUT3 was more significant (P <0.01), but no dose-related relationship. Conclusion The model of neurotoxicity caused by methamphetamines was established. Methamphetamine had a characteristic and dose-dependent effect on the behavior of zebrafish.