Citation:

Hill, B., K. Britton, D. Hunter, J. Olin, M. Lowery, J. Hedge, B. Knapp, K. Jarema, Z. Rowson, AND S. Padilla. Inconsistencies in Variable Reporting and Methods in Larval Zebrafish Behavioral Assays. NEUROTOXICOLOGY AND TERATOLOGY. Elsevier Science Ltd, New York, NY, 96:107163, (2023). https://doi.org/10.1016/j.ntt.2023.107163

Impact/Purpose:

New consistent approaches in developmental neurotoxicity screening are needed, but without a standardized protocol for behavioral assays, comparison of results among laboratories is challenging. To determine how congruent protocols are across laboratories, we conducted a literature review of developmental neurotoxicity studies focusing on zebrafish larval behavior assays to catalog experimental design consistencies. We found that while there were some aspects of the experimental design that were consistent among laboratories, none were the same across any of the publications, exposing considerable inconsistencies among methods. There is a strong need for the development of (1) a standardized testing protocol for larval zebrafish locomotor assays and (2) a standardized protocol for reporting experimental variables in the literature. We have also developed an extensive guideline checklist for reporting the conduct of larval zebrafish developmental assays.

Description:

New approaches in developmental neurotoxicity (DNT) screening are needed due to the tens of thousands of chemicals requiring hazard assessments. Zebrafish (Danio rerio) are an alternative vertebrate model for DNT testing, but without a standardized protocol for larval behavioral assays, comparison of results among laboratories is challenging. To evaluate the congruence of protocols across laboratories, we conducted a literature review of DNT studies focusing on larval zebrafish behavior assays and cataloged experimental design consistencies. Our review focused on 51 unique method variables in publications where chemical exposure occurred in early development and subsequent larval locomotor evaluation focused on assays that included a light/dark photoperiod transition. We initially identified 94 publications, but only 31 exclusively met our inclusion criteria which focused on parameters that are important to an assay employed by our laboratory. No publication reported 100% of the targeted variables; only 51 to 86% of those variables were reported in the reviewed publications, with some aspects of the experimental design consistent among laboratories. However, no protocol was exactly the same for any two publications. Many of these variables had more than one parameter/design reported, highlighting the inconsistencies among methods. Overall, there is not only a strong need for the development of a standardized testing protocol for larval zebrafish locomotor assays, but there is also a need for a standardized protocol for reporting experimental variables in the literature. Here we include an extensive guideline checklist for conducting larval zebrafish developmental behavior assays.

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