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Inconsistencies in variable reporting and methods in larval zebrafish behavioral assays.

Bridgett N Hill1, Katy N Britton2, Deborah L Hunter3

  • 1ORISE Research Participation Program hosted by EPA, Center for Computational Toxicology and Exposure, Biomolecular and Computational Toxicology Division, Rapid Assay Development Branch, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA.

Neurotoxicology and Teratology
|February 9, 2023
PubMed
Summary

Standardizing larval zebrafish behavior assays is crucial for reliable developmental neurotoxicity (DNT) screening. This study highlights inconsistencies in current DNT testing protocols and advocates for standardized methods and reporting.

Keywords:
Assay confoundersBehaviorDevelopmental neurotoxicityLarvalReporting recommendationsZebrafish

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Area of Science:

  • Toxicology
  • Neuroscience
  • Ecotoxicology

Background:

  • Developmental neurotoxicity (DNT) screening requires new approaches due to the vast number of chemicals needing assessment.
  • Zebrafish (Danio rerio) offer a vertebrate alternative for DNT testing, but a lack of standardized larval behavioral assays hinders inter-laboratory comparisons.

Purpose of the Study:

  • To evaluate the congruence of existing larval zebrafish behavioral assay protocols across different laboratories.
  • To identify inconsistencies in experimental designs and reporting within DNT studies using zebrafish.

Main Methods:

  • Conducted a literature review of 94 DNT studies focusing on larval zebrafish behavior assays with light/dark photoperiod transitions.
  • Cataloged 51 unique method variables related to experimental design and chemical exposure.
  • Included only 31 publications that met specific inclusion criteria for detailed analysis.

Main Results:

  • No single publication reported all targeted variables; reported variables ranged from 51% to 86%.
  • While some experimental design aspects were consistent, no two publications shared an identical protocol.
  • Significant inconsistencies were found in multiple reported method variables, indicating a lack of standardization.

Conclusions:

  • There is a critical need for standardized testing protocols for larval zebrafish locomotor assays in DNT screening.
  • Standardized reporting guidelines for experimental variables in DNT literature are also essential.
  • An extensive guideline checklist is provided to aid in conducting standardized larval zebrafish developmental behavior assays.