Brewing Anxiety: Caffeine‐Induced Thigmotaxis in Zebrafish Larvae
- 1Department of Molecular Biology, Cell Biology and Biochemistry Brown University Providence Rhode Island USA
Abstract
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.
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BibTeX
@article{rai2026brewing,
title = {{Brewing Anxiety: Caffeine‐Induced Thigmotaxis in Zebrafish Larvae}},
author = {Rishi Rai and Thais Del Rosario Hernandez and Robbert Creton},
journal = {Brain and Behavior},
year = {2026},
month = oct,
doi = {10.1002/brb3.71769},
url = {https://doi.org/10.1002/brb3.71769},
note = {CC BY},
}
RIS
TY - JOUR
TI - Brewing Anxiety: Caffeine‐Induced Thigmotaxis in Zebrafish Larvae
AU - Rishi Rai
AU - Thais Del Rosario Hernandez
AU - Robbert Creton
PY - 2026
DA - 2026/10/25
JO - Brain and Behavior
DO - 10.1002/brb3.71769
UR - https://doi.org/10.1002/brb3.71769
AB - 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.
SN - 2162-3279
ER -