National Wildlife Research Centre, Environment and Climate Change Canada, Ottawa, Ontario, Canada.
Department of Natural Resource Sciences, McGill University, Montreal, Quebec, Canada.
Environ Toxicol Chem. 2023 Aug;42(8):1763-1771. doi: 10.1002/etc.5676. Epub 2023 Jun 7.
New approach methods (NAMs) are increasingly important to help accelerate the pace of ecological risk assessment and offer more ethical, affordable, and efficient alternatives to traditional toxicity tests. In the present study, we describe the development, technical characterization, and initial testing of a toxicogenomics tool, EcoToxChip (384-well quantitative polymerase chain reaction [qPCR] array), to support chemical management and environmental monitoring for three laboratory model species-fathead minnow (Pimephales promelas), African clawed frog (Xenopus laevis), and Japanese quail (Coturnix japonica). Chip design, including gene selection, was informed by a diverse end-user group and quality control metrics (e.g., primer assay, reverse transcription, and PCR efficiency) performed well based on a priori established criteria. Correlation with RNA sequencing (seq) data provided additional confidence in this novel toxicogenomics tool. Although the present study represents an initial testing of only 24 EcoToxChips for each of the model species, the results provide increased confidence in the robustness/reproducibility of EcoToxChips for evaluating perturbations in gene expression associated with chemical exposure and thus, this NAM, combined with early-life stage toxicity testing, could augment current efforts for chemical prioritization and environmental management. Environ Toxicol Chem 2023;42:1763-1771. © 2023 SETAC.
新方法(NAMs)对于加快生态风险评估的步伐非常重要,它们为传统毒性测试提供了更具道德、更经济、更有效的替代方法。本研究描述了一种毒理基因组学工具——EcoToxChip(384 孔定量聚合酶链反应 [qPCR] 阵列)的开发、技术特征和初步测试,以支持三种实验室模式物种——黑头呆鱼(Pimephales promelas)、非洲爪蟾(Xenopus laevis)和日本鹌鹑(Coturnix japonica)的化学管理和环境监测。芯片设计,包括基因选择,是由一个多样化的最终用户群体提供信息,并根据事先确定的标准,通过质量控制指标(例如,引物检测、逆转录和 PCR 效率)进行了很好的评估。与 RNA 测序(seq)数据的相关性为这种新的毒理基因组学工具提供了额外的可信度。尽管本研究仅代表对每个模型物种的 24 个 EcoToxChips 进行了初步测试,但结果提高了对 EcoToxChips 评估与化学暴露相关的基因表达扰动的稳健性/可重复性的信心,因此,这种新方法与早期生活阶段毒性测试相结合,可以增强当前用于化学优先排序和环境管理的努力。环境毒理化学 2023;42:1763-1771。 © 2023 SETAC。