Comparative Analyses of Zebrafish Anxiety-Like Behavior Using Conflict-Based Novelty Tests

Elana V. Kysil, Darya A. Meshalkina, Erin E. Frick, David J. Echevarria, Denis B. Rosemberg, Caio Maximino, Monica Gomes Lima, Murilo S. Abreu, Ana C. Giacomini, Leonardo J.G. Barcellos, Cai Song, Allan V. Kalueff*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

155 Citations (Scopus)


Modeling of stress and anxiety in adult zebrafish (Danio rerio) is increasingly utilized in neuroscience research and central nervous system (CNS) drug discovery. Representing the most commonly used zebrafish anxiety models, the novel tank test (NTT) focuses on zebrafish diving in response to potentially threatening stimuli, whereas the light-dark test (LDT) is based on fish scototaxis (innate preference for dark vs. bright areas). Here, we systematically evaluate the utility of these two tests, combining meta-analyses of published literature with comparative in vivo behavioral and whole-body endocrine (cortisol) testing. Overall, the NTT and LDT behaviors demonstrate a generally good cross-test correlation in vivo, whereas meta-analyses of published literature show that both tests have similar sensitivity to zebrafish anxiety-like states. Finally, NTT evokes higher levels of cortisol, likely representing a more stressful procedure than LDT. Collectively, our study reappraises NTT and LDT for studying anxiety-like states in zebrafish, and emphasizes their developing utility for neurobehavioral research. These findings can help optimize drug screening procedures by choosing more appropriate models for testing anxiolytic or anxiogenic drugs.

Original languageEnglish
Pages (from-to)197-208
Number of pages12
Issue number3
Publication statusPublished - Jun 2017
Externally publishedYes


  • anxiety-like behavior
  • behavioral phenotyping
  • the light-dark test
  • the novel tank test
  • Zebrafish


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