Refinement of the zebrafish embryo developmental toxicity assay.
Refinement of the zebrafish embryo developmental toxicity assay.
复制标题
斑马鱼胚胎发育毒性测定法的细化。
DOI：
10.1016/j.mex.2020.101087
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发表时间：
2020
期刊：
影响因子：
1.9
通讯作者：
Van Cruchten S
中科院分区：
文献类型：
--
作者：
Hoyberghs J;Bars C;Pype C;Foubert K;Ayuso Hernando M;Van Ginneken C;Ball J;Van Cruchten S
我们的方案是使用人肝微粒体定制的，具有用于测试化合物的代谢活化系统。该系统确保斑马鱼胚胎暴露于与人类风险和安全评估相关的代谢物。由于人肝微粒体对斑马鱼胚胎具有毒性，我们开发了一种具有超离心和随后稀释步骤的预孵育系统。此外，我们开发了一种骨骼染色方案，可添加到ZEDTA模块化系统中。我们的活体茜素红染色方法以一致的方式检测了5天龄斑马鱼幼鱼的几种骨结构。一些制药和化学公司正在使用斑马鱼胚胎作为动物试验的替代品，用于早期检测发育毒物。不幸的是，这种斑马鱼胚胎试验的方案在实验室之间存在差异，导致相同化合物的数据不一致。与体内哺乳动物发育毒性研究相比，该试验也有一些局限性，如生物转化能力低和形态学终点少。因此，需要标准化和进一步优化发育毒性试验的测定。我们开发了一种斑马鱼胚胎发育毒性试验（ZEDTA），可以用代谢活化系统和/或骨骼染色来扩展，以增加其灵敏度。因此，ZEDTA可以用作模块化系统，这取决于感兴趣的化合物。
Our protocol is customized with a metabolic activation system for test compounds, using human liver microsomes. This system ensures exposure of zebrafish embryos to metabolites that are relevant for human risk and safety assessment. As human liver microsomes are toxic for the zebrafish embryo, we developed a preincubation system with an ultracentrifugation and subsequent dilution step. Additionally, we developed a skeletal staining protocol that can be added to the ZEDTA modular system. Our live Alizarin Red staining method detects several bone structures in 5-day old zebrafish larvae in a consistent manner. Several pharmaceutical and chemical companies are using the zebrafish embryo as an alternative to animal testing for early detection of developmental toxicants. Unfortunately, the protocol of this zebrafish embryo assay varies between labs, resulting in discordant data for identical compounds. The assay also has some limitations, such as low biotransformation capacity and fewer morphological endpoints in comparison with the in vivo mammalian developmental toxicity studies. Consequently, there is a need to standardize and further optimize the assay for developmental toxicity testing. We developed a Zebrafish Embryo Developmental Toxicity Assay (ZEDTA) that can be extended with a metabolic activation system and/or skeletal staining to increase its sensitivity. As such, the ZEDTA can be used as a modular system depending on the compound of interest.
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影响因子：
3.3
作者：
Gustafson, A. -L.;Stedman, D. B.;Augustine-Rauch, K. A.
通讯作者：
Augustine-Rauch, K. A.
影响因子：
5.6
作者：
Giusti, Arianna;Xuan-Bac Nguyen;de Witte, Peter
通讯作者：
de Witte, Peter
影响因子：
3.3
作者：
Mattsson, Anna;Ulleras, Erik;Oskarsson, Agneta
通讯作者：
Oskarsson, Agneta
影响因子：
--
作者：
Bensimon-Brito, A.;Cardeira, J.;Witten, P. E.
通讯作者：
Witten, P. E.
影响因子：
3.8
作者：
Ball, Jonathan S.;Stedman, Donald B.;Augustine-Rauch, Karen A.
通讯作者：
Augustine-Rauch, Karen A.