The sensitivity of the zebrafish embryo coiling assay for the detection of neurotoxicity by compounds with diverse modes-of-action

Abstract In the aim to determine neurotoxicity, new methods are being validated, including tests and test batteries comprising in vitro and in vivo approaches. Alternative test models such as the zebrafish (Danio rerio) embryo have received increasing attention, with minor modifications of the fish embryo toxicity test (FET; OECD TG 236) as a tool to assess behavioral endpoints related to neurotoxicity during early developmental stages. The spontaneous tail movement assay, also known as coiling assay, assesses the development of random movement into complex behavioral patterns and has proven sensitive to acetylcholine esterase inhibitors at sublethal concentrations. The present study explored the sensitivity of the assay to neurotoxicants with other modes-of-action. Here, five compounds with diverse modes-of-action were tested at sublethal concentrations: acrylamide, carbaryl, hexachlorophene, ibuprofen, and rotenone. While carbaryl, hexachlorophene, and rotenone consistently induced severe behavioral alterations by ~30 hours post fertilization (hpf), acrylamide and ibuprofen expressed time- and/or concentration-dependent effects. At 37 - 38 hpf, additional observations revealed behavioral changes during dark phases with a strict concentration-dependency. The study documented the applicability of the coiling assay to mode-of-action-dependent behavioral alterations at sublethal concentrations, underlining its potential as a component of a neurotoxicity test battery.