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

Rebecca von Hellfeld* (Corresponding Author), Christoph Andreas Gade, Lisa Baumann, Marcel Leist, Thomas Braunbeck

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


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.
Original languageEnglish
Pages (from-to)75281–75299
Number of pages19
JournalEnvironmental Science and Pollution Research
Early online date22 May 2023
Publication statusPublished - 1 Jun 2023

Bibliographical note

Open Access via the Springer Agreement
Funding: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 681002 (EU-ToxRisk).

Data Availability Statement

Availability of data and materials
Original datasets of the current study and analyses generated are available in the BioStudies repository (


  • locomotor assay
  • spontaneous tail movement
  • Danio rerio
  • behavior profiling
  • developmental toxicity testing
  • alternative test method


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