Department of Environmental Science, Stockholm University, Sweden.
Mol Omics. 2020 Jun 1;16(3):185-194. doi: 10.1039/d0mo00007h. Epub 2020 Mar 19.
Toxicometabolomic studies involving zebrafish embryos have become increasingly popular for linking apical endpoints to biochemical perturbations as part of adverse outcome pathway determination. These experiments involve pooling embryos to generate sufficient biomass for metabolomic measurement, which adds both time and cost. To address this limitation, we developed a high-throughput toxicometabolomic assay involving single zebrafish embryos. Incubation, microscopy, embryo extraction, and instrumental metabolomic analysis were all performed in the same 96-well plate, following acquisition of conventional toxicological endpoints. The total time for the assay (including testing of 6 doses/n = 12 embryos per dose plus positive and negative controls, assessing conventional endpoints, instrumental analysis, data processing and multivariate statistics) is <14 days. Metabolomic perturbations at low dose were linked statistically to those observed at high dose and in the presence of an adverse effect, thereby contextualizing omic data amongst apical endpoints. Overall, this assay enables collection of high resolution metabolomic data in a high throughput manner, suitable for mode of action hypothesis generation in the context of pharmaceutical or toxicological screening.
毒代代谢组学研究涉及斑马鱼胚胎，已越来越多地用于将顶端终点与生化干扰联系起来，作为不良结局途径确定的一部分。这些实验涉及到汇集胚胎以产生足够的生物量进行代谢组学测量，这增加了时间和成本。为了解决这个限制，我们开发了一种涉及单个斑马鱼胚胎的高通量毒代代谢组学测定法。孵育、显微镜检查、胚胎提取和仪器代谢组学分析都在同一个 96 孔板中进行，同时获得常规毒理学终点。该测定法的总时间（包括测试 6 个剂量/n = 每个剂量 12 个胚胎加上阳性和阴性对照，评估常规终点，仪器分析，数据处理和多元统计）<14 天。低剂量的代谢组学干扰与高剂量和存在不良效应时观察到的干扰在统计学上相关联，从而将组学数据置于顶端终点之间。总的来说，该测定法能够以高通量的方式收集高分辨率代谢组学数据，适用于药物或毒理学筛选背景下作用模式假设的生成。
