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Screen for Developmental Toxicity in Zebrafish- Study 1

Celia Quevedo, Mamta Behl, Kristen Ryan, Richard S. Paules, Aintzane Alday, Arantza Muriana, and Ainhoa Alzualde. 2019. Detection and Prioritization of Developmentally Neurotoxic and/or Neurotoxic Compounds Using Zebrafish. TOXICOLOGICAL SCIENCES, 168(1), 2019, 225–240. doi: 10.1093/toxsci/kfy291

DOI: https://doi.org/10.22427/NTP-DATA-002-00077-0001-0000-7

Materials and Methods

Study Description

A full description of the materials and methods can be found in Quevedo et al., 2019. All experiments were conducted at Biobide (Donostia-San Sebastián, Spain). Eighty-seven chemicals tested were provided by the National Toxicology Program (Research Triangle Park, NC, USA) and are cited in Quevdo et al., 2019. An additional chemical, 2-((1-(4-Phenoxyphenoxy)propan-2-yl)oxy)pyridine: MPEP (CASRN: 95737-68-1) was evaluated in parallel and thus there are results for a total of 88 chemicals. In brief, the adult zebrafish line Tg(Cmlc2: copGFP) was used to evaluate embryo toxicity following chemical exposure. Eighty-eight compounds were evaluated in dose response with ~8 concentrations of each chemical. Fertilized embryos, at 4 hours post fertilization, were placed in 24-well plates with the corresponding chemical concentration or 0.5% DMSO as a negative control. The plates were covered and wrapped with aluminum foil to avoid degradation of light-sensitive compounds. Fifteen embryos were analyzed per condition after 2 and 4 days of incubation (i.e. 2 and 4 days post fertilization (dpf)) at 28.5°C. Detailed analysis of embryo morphology (including malformations in the head, heart and tail, deformed body shape, and the presence of edemas) and lethality was performed at 2 and 4 dpf. Embryo morphology was visualized under a stereo microscope (Olympus SXZ10) by experienced technicians. Morphologies found on study are recorded as present (1) or absent (0).