冈田酸对斑马鱼幼鱼神经行为功能的影响

Impacts of Okadaic Acid on Neurological Function and Behavior of Zebrafish Larvae

目的研究冈田酸(okadaic acid,OA)对斑马鱼幼鱼神经行为功能的影响,初步探索斑马鱼模型在神经退行性疾病研究中的应用前景。方法用0.05、0.10和0.20μmol/L的OA溶液处理受精后3 d(3 dpf)的斑马鱼幼鱼7 d,记录幼鱼死亡数,并利用行为学分析系统采集幼鱼30 min内的游动轨迹,测量每组幼鱼的游动距离和呆滞时间。采用实时荧光定量PCR法检测各处理组幼鱼体内蛋白磷酸酶2A(PP2A)和α7烟碱型乙酰胆碱受体(α7-nAChR)的表达情况。结果与对照组比较,0.05μmol/L和0.10μmol/L OA组处理7 d后的幼鱼存活率无显著差异(P值均>0.05),游动轨迹也无明显改变。与对照组比较,0.20μmol/L OA组幼鱼存活率明显降低,游动活力明显下降,游动距离明显缩短,呆滞时间明显延长(P值均<0.05);而且幼鱼体内PP2A基因表达水平明显下调(P<0.05),α7-nAChR基因表达水平则无明显改变(P>0.05)。结论0.20μmol/L的OA处理7 d导致斑马鱼幼鱼的神经行为功能异常。

Objective To investigate the impacts of okadaic acid(OA)on the neurological function and behavior of zebrafish larvae,and explore the possible application of zebrafish as an animal model in the study of neurodegenerative diseases.Methods Zebrafish larvae at 3 days post fertilization(3 dpf)stage was treated with 0.05μmol/L,0.10μmol/L,and 0.20μmol/L OA respectively for 7 days.The survival rate of zebrafish larvae was calculated,the swimming track within 30 min was acquired by behaviroral analysis system,and the swimming trajectory,swimming distance and inactive time were recorded.The expression levels of protein phosphatase 2 A(PP2A)and nicotinic acetylcholine receptor alpha 7(α7-nAChR)genes were tested by real-time fluorescent quantitative PCR.Results After 7 days of treatment,the survival rate and swimming trajectory of the zebrafish larvae in 0.05μmol/L and 0.10μmol/L OA treatment groups were not significantly different from those in the control group(P>0.05).Zebrafish larvae treated with 0.20μmol/L OA for 7 days showed inactive swimming,reduced swimming distance and prolonged sluggish time with significant difference as compared with the control group(P<0.05).The expression level of PP2A gene was down-regulated significantly in 0.20μmol/L OA treatment group(P<0.05),however the expression level ofα7-nAChR was not affected(P>0.05).Conclusion The treatment with 0.20μmol/L OA for 7 days can cause the abnormal neurological function and behavior of zebrafish larvae.