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Updated: Oct 1, 2026
Automated High-throughput Behavioral Analyses in Zebrafish Larvae
Published on: July 4, 2013
Brewing Anxiety: Caffeine-Induced Thigmotaxis in Zebrafish Larvae
Rishi Rai1, Thais Del Rosario Hernandez1, Robbert Creton1
1Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island, USA.
Background:
Thigmotaxis, or edge preference, is widely used as a measure of anxiety in vertebrate model systems. However, variability in this behavior limits its utility as a scalable measure for anxiolytic drug discovery. In the current study, we examine caffeine-induced thigmotaxis, using 5-day-old zebrafish larvae as a model system.
Method:
We modified our imaging system and AI-assisted analysis for measurements of behavior in six-well plates. The imaging system was used to measure thigmotaxis and several other behaviors, including overall activity, scoot and burst swimming, and responses to acoustic and visual stimuli.
Result:
Caffeine (100 mg/L) caused the zebrafish larvae to exhibit increased edge preference, decreased activity, increased scooting, and decreased bursting behavior. Coadministration with diazepam (0.25 mg/L), a central nervous system depressant, significantly reduced caffeine-induced thigmotaxis and increased locomotor activity.
Conclusion:
These findings support edge preference as a reliable measure of anxiety and reconcile existing conflicting results regarding diazepam's anxiolytic efficacy in zebrafish larvae, suggesting that anxiolytic effects are most pronounced against a heightened anxiety baseline. These findings also establish a robust platform for high-throughput identification of anxiolytic compounds with potential relevance for treating anxiety and related stress in humans.
