PUBLICATION

A unified approach to investigating 4 dpf zebrafish larval behaviour through a standardised light/dark assay

Authors
Hillman, C., Kearn, J., Parker, M.O.
ID
ZDB-PUB-240714-6
Date
2024
Source
Progress in neuro-psychopharmacology & biological psychiatry   134: 111084 (Journal)
Registered Authors
Parker, Matt
Keywords
3Rs of animal research, Light/dark transition test, Locomotor activity, Standardised testing, Zebrafish
MeSH Terms
  • Photoperiod
  • Central Nervous System Stimulants/pharmacology
  • Motor Activity/drug effects
  • Motor Activity/physiology
  • Animals, Genetically Modified
  • Locomotion*/drug effects
  • Locomotion*/physiology
  • Behavior, Animal*/drug effects
  • Behavior, Animal*/physiology
  • Zebrafish*/physiology
  • Reproducibility of Results
  • Animals
  • Ethanol/pharmacology
  • Larva*
  • Light
PubMed
39002928 Full text @ Prog. Neuropsychopharmacol. Biol. Psychiatry
Abstract
Zebrafish have emerged as a dynamic research model in the domains of neuropsychopharmacology, biological psychiatry and behaviour. Working with larvae ≤4 days post-fertilisation (dpf) offers an avenue for high-throughput investigation whilst aligning with the 3Rs principles of animal research. The light/dark assay, which is the most used behavioural assay for larval neuropharmacology research, lacks experimental reliability and standardisation. This study aimed to formulate a robust, reproducible and standardised light/dark behavioural assay using 4 dpf zebrafish larvae. Considerable between-batch and inter-individual variability was found, which we rectified with a normalisation approach to ensure a reliable foundation for analysis. We then identified that 5-min light/dark transition periods are optimal for locomotor activity. We also found that a 30-min acclimation in the light was found to produce significantly increased dark phase larval locomotion. Next, we confirmed the pharmacological predictivity of the standardised assay using ethanol which, as predicted, caused hyperlocomotion at low concentrations and hypolocomotion at high concentrations. Finally, the assay was validated by assessing the behavioural phenotype of hyperactive transgenic (adgrl3.1-/-) larvae, which was rescued with psychostimulant medications. Our standardised assay not only provides a clear experimental and analytical framework to work with 4 dpf larvae, but also facilitates between-laboratory collaboration using our normalisation approach.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
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Mapping