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使用斑马鱼胚胎快速评估化合物毒性

Rapid Evaluation of Toxicity of Chemical Compounds Using Zebrafish Embryos.

作者信息

Aspatwar Ashok, Hammaren Milka Marjut, Parikka Mataleena, Parkkila Seppo

机构信息

Faculty of Medicine and Health Technology, Tampere University;

Faculty of Medicine and Health Technology, Tampere University.

出版信息

J Vis Exp. 2019 Aug 25(150). doi: 10.3791/59315.

Abstract

The zebrafish is a widely used vertebrate model organism for the disease and phenotype-based drug discovery. The zebrafish generates many offspring, has transparent embryos and rapid external development. Zebrafish embryos can, therefore, also be used for the rapid evaluation of toxicity of the drugs that are precious and available in small quantities. In the present article, a method for the efficient screening of the toxicity of chemical compounds using 1-5-day post fertilization embryos is described. The embryos are monitored by stereomicroscope to investigate the phenotypic defects caused by the exposure to different concentrations of compounds. Half-maximal lethal concentrations (LC50) of the compounds are also determined. The present study required 3-6 mg of an inhibitor compound, and the whole experiment takes about 8-10 h to be completed by an individual in a laboratory having basic facilities. The current protocol is suitable for testing any compound to identify intolerable toxic or off-target effects of the compound in the early phase of drug discovery and to detect subtle toxic effects that may be missed in the cell culture or other animal models. The method reduces procedural delays and costs of drug development.

摘要

斑马鱼是一种广泛用于基于疾病和表型的药物发现的脊椎动物模式生物。斑马鱼繁殖后代众多,胚胎透明且外部发育迅速。因此,斑马鱼胚胎也可用于对珍贵且数量稀少的药物进行毒性快速评估。在本文中,描述了一种使用受精后1至5天的胚胎高效筛选化合物毒性的方法。通过体视显微镜监测胚胎,以研究暴露于不同浓度化合物所引起的表型缺陷。还确定了化合物的半数致死浓度(LC50)。本研究需要3至6毫克的抑制剂化合物,在具备基本设施的实验室中,一个人完成整个实验大约需要8至10小时。当前方案适用于测试任何化合物,以在药物发现的早期阶段识别该化合物无法耐受的毒性或脱靶效应,并检测在细胞培养或其他动物模型中可能遗漏的细微毒性效应。该方法减少了药物开发过程中的延误和成本。

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