Institute of Toxicology, Merck KGaA, 64293 Darmstadt, Germany.
Toxicology. 2011 Mar 15;281(1-3):25-36. doi: 10.1016/j.tox.2011.01.004. Epub 2011 Jan 13.
Zebrafish embryos have been shown to be a useful model for the detection of direct acting teratogens. This communication presents a protocol for a 3-day in vitro zebrafish embryo teratogenicity assay and describes results obtained for 10 proteratogens: 2-acetylaminofluorene, benzo[a]pyrene, aflatoxin B(1), carbamazepine, phenytoin, trimethadione, cyclophosphamide, ifosfamide, tegafur and thio-TEPA. The selection of the test substances accounts for differences in structure, origin, metabolism and water solubility. Apart from 2-acetylaminofluorene, which mainly produces lethal effects, all proteratogens tested were teratogenic in zebrafish embryos exposed for 3 days. The test substances and/or the substance class produced characteristic patterns of fingerprint endpoints. Several substances produced effects that could be identified already at 1 dpf (days post fertilization), whereas the effects of others could only be identified unambiguously after hatching at ≥ 3 dpf. The LC₅₀ and EC₅₀ values were used to calculate the teratogenicity index (TI) for the different substances, and the EC₂₀ values were related to human plasma concentrations. Results lead to the conclusion that zebrafish embryos are able to activate proteratogenic substances without addition of an exogenous metabolic activation system. Moreover, the teratogenic effects were observed at concentrations relevant to human exposure data. Along with other findings, our results indicate that zebrafish embryos are a useful alternative method for traditional teratogenicity testing with mammalian species.
斑马鱼胚胎已被证明是检测直接作用致畸物的有用模型。本通讯介绍了一种为期 3 天的体外斑马鱼胚胎致畸性测定法，并描述了 10 种原致畸物的结果：2-乙酰氨基芴、苯并[a]芘、黄曲霉毒素 B(1)、卡马西平、苯妥英、三甲双酮、环磷酰胺、异环磷酰胺、替加氟和硫代四嗪。测试物质的选择考虑了结构、来源、代谢和水溶性的差异。除了主要产生致死作用的 2-乙酰氨基芴外，所有测试的原致畸物在暴露 3 天的斑马鱼胚胎中均具有致畸作用。测试物质和/或物质类别产生了特征性的指纹终点模式。几种物质产生的效应在 1 dpf（受精后天数）时即可识别，而其他物质的效应只能在孵出后≥3 dpf 时才能明确识别。LC₅₀和 EC₅₀ 值用于计算不同物质的致畸指数 (TI)，EC₂₀ 值与人类血浆浓度相关。结果得出结论，斑马鱼胚胎能够激活原致畸物质，而无需添加外源性代谢激活系统。此外，在与人类暴露数据相关的浓度下观察到了致畸作用。结合其他发现，我们的结果表明，斑马鱼胚胎是传统致畸性测试的哺乳动物替代方法。
