Publication Details |
| Category | Text Publication |
| Reference Category | Journals |
| DOI | 10.1289/EHP16568 |
| Title (Primary) | Multi-behavioral phenotyping in early-life-stage zebrafish for identifying disruptors of non-associative learning |
| Author | Leuthold, D.; Herold, N.K.; Nerlich, J.; Bartmann, K.; Scharkin, I.; Hallermann, S.J.; Schweiger, N.; Fritsche, E.; Tal, T.
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| Source Titel | Environmental Health Perspectives |
| Year | 2025 |
| Department | ETOX |
| Language | englisch |
| Topic | T9 Healthy Planet |
| Supplements | https://ehp.niehs.nih.gov/doi/suppl/10.1289/EHP16568/suppl_file/ehp16568.smcontents.508.pdf https://ehp.niehs.nih.gov/doi/suppl/10.1289/EHP16568/suppl_file/ehp16568.s001.acco.pdf https://ehp.niehs.nih.gov/doi/suppl/10.1289/EHP16568/suppl_file/ehp16568.s002.codeanddata.acco.zip |
| Abstract | Background: The vertebrate nervous system is vulnerable to chemical toxicity and the widespread release of chemicals into the environment outstrips the capacity to assess their safety. The zebrafish (Danio rerio) is a powerful vertebrate model that can bridge the gap between in vitro and mammalian-based in vivo studies. However, the behavior-rich repertoire of larval zebrafish, a 3R-compliant model amenable to higher-throughput chemical screens, has yet to be fully deployed to identify and characterize chemical compounds that cause neurotoxicity. Objective: We sought to establish a multi-behavioral phenotyping approach in larval zebrafish to identify and mechanistically elucidate neuroactive chemicals, with particular focus on chemical compounds that affect non-associative habituation learning. Methods: We devised a battery of automated behavior assays in larval zebrafish. The battery captures stereotypical visual and acoustic behaviors including habituation, a form of non-associative learning. To elucidate mechanisms underlying exposure-induced behavioral alterations in zebrafish, in silico target predictions, pharmacological interventions, patch-clamp recordings in cultured mouse cortical neurons, and human multi-neurotransmitter (hMNR) assay in 3D BrainSpheres were used. Results: Known pharmacological modulators of habituation in zebrafish evoked distinct behavioral patterns. By screening chemicals positive for ex vivo N-methyl-D-aspartate receptor (NMDAR) modulation, we identified chlorophene, a biocide that caused sedation, paradoxical excitation, and reduced habituation in zebrafish. Using in silico target predictions and pharmacological interventions, we discovered that chlorophene acts via gamma-aminobutyric acid A receptors (GABAARs), a previously unknown target site. Orthogonal validation in cultured mouse cortical neurons and human stem cell-derived BrainSpheres confirmed chlorophene’s interaction with GABAARs. Chlorophene’s behavioral profile resembled that of flupirtine, a Kv7 potassium channel (M-current) activator, suggesting that habituation deficits stem from M-current rather than GABAAR modulation. Conclusions: These studies combined a series of behavior assays in a phenotypically rich, rapid, and inexpensive non-mammalian vertebrate test system to screen chemicals for neurotoxicity. Together with in silico target predictions and mouse- and human-based models, our findings establish multi-behavioral phenotyping in zebrafish as a powerful toolkit for neurotoxicity testing and mechanism identification, with relevance for humans. |
| Persistent UFZ Identifier | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=29721 |
| Leuthold, D., Herold, N.K., Nerlich, J., Bartmann, K., Scharkin, I., Hallermann, S.J., Schweiger, N., Fritsche, E., Tal, T. (2025): Multi-behavioral phenotyping in early-life-stage zebrafish for identifying disruptors of non-associative learning Environ. Health Perspect. 10.1289/EHP16568 |
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