实验动物与比较医学 ›› 2020, Vol. 40 ›› Issue (3): 190-.DOI: 10.3969/j.issn.1674-5817.2020.03.003

• 论著 • 上一篇    下一篇

冈田酸对斑马鱼幼鱼神经行为功能的影响

王   雪1,2, 刘可春1,2, 杨学亮1, 马玉奎3, 张   云1,2    

  1. 1. 齐鲁工业大学(山东省科学院)生物研究所, 济南 250103; 
    2. 山东省人类疾病斑马鱼模型与药物筛选工程技术研究中心, 济南 250103;
    3. 山东省药学科学院, 山东省化学药物重点实验室, 济南 250101
  • 收稿日期:2019-11-11 出版日期:2020-06-25 发布日期:2020-12-16
  • 作者简介:王 雪(1978—), 女, 硕士, 副研究员, 研究方向: 药物活性筛选与评价。E-mail: wangxue8809@163.com
  • 基金资助:
    国家重点研发计划项目(2018YFC1707300)

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

WANG Xue1,2, LIU Kechun1,2, YANG Xueliang1, MA Yukui3, ZHANG Yun1,2   

  1. 1. Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250103, China;  2. Engineering Research Center of Zebrafish Models for Human Diseases and Drug Screening of Shandong Province, Jinan 250103, China;  3. Shandong Provincial Key Laboratory of Chemical Drugs, Shandong Academy of Pharmaceutical Sciences, Jinan 250101, China
  • Received:2019-11-11 Online:2020-06-25 Published:2020-12-16

摘要: 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0.20 µmol/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,0.05 µmol/L 和0.10 µmol/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,0.20 µmol/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    0.20 µmol/L的OA处理7 d导致斑马鱼幼鱼的神经行为功能异常。


关键词: 斑马鱼, 冈田酸, 神经退行性疾病, 碱性磷酸酶2A, α7烟碱型乙酰胆碱受体

Abstract:  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 0.05 µmol/L, 0.10 µmol/L, and 0.20 µmol/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 2A (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 0.05 µmol/L and 0.10 µmol/L OA treatment groups were not significantly different from those in the control group (P>0.05). Zebrafish larvae treated with 0.20 µmol/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 0.20 µmol/L OA treatment group (P<0.05), however the expression level of α7-nAChR was not affected (P>0.05).  Conclusion   The treatment with 0.20 µmol/L OA for 7 days can cause the abnormal neurological function and behavior of zebrafish larvae. 

Key words: Zebrafish, Okadaic acid, Neurodegenerative disease, Protein phosphatase 2A; Nicotinic acetylcholine receptor alpha 7