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Effects of Larval Age and Previous Testing on Larval Zebrafish Behavior
Citation:
Barbo, N., D. Hunter, B. Knapp, AND S. Padilla. Effects of Larval Age and Previous Testing on Larval Zebrafish Behavior. North Carolina Society of Toxicology (NCSOT) Annual Meeting September 2023, Research Triangle Park, NC, September 14, 2023. https://doi.org/10.23645/epacomptox.24156246
Impact/Purpose:
Presentation to the North Carolina Society of Toxicology (NCSOT) Annual Meeting September 2023
Description:
Zebrafish (Danio rerio) are often used to investigate the effects of chemical exposure during development. Brain development in the zebrafish larval stage is rapid, so it is common for larvae to be assessed at various developmental stages, such as 4, 5, and 6 days post fertilization (dpf). Chemical screening and assay development can benefit from repeated behavioral assays on the same individuals over consecutive days because such experiments allow for a better understanding of exposure over time. With multiple timepoints being assessed using the same fish, it is important to understand: (1) how larval age affects locomotor behavior and (2) how previous behavior testing impacts later behavior profiles. To accomplish this, we concurrently reared two 96-well plates of zebrafish embryos/larvae at 26°C (one individual per well, experiment repeated 3x) and conducted a light-dark transition test on larvae at both 4 and 5 dpf, or only 5 dpf. When comparing behavior at 4 to 5 dpf (n=180), 4 dpf fish moved significantly less, especially in the light phase (-90%). When assessing whether prior testing affected behavior, there was a significant but small (9%) decrease in locomotion in the dark phase among individuals who experienced previous testing (n=279-281). This decreased movement in previously tested fish, although statistically significant, is within normal behavior variability and is unlikely to impact experimental outcomes when using a smaller number of individuals per treatment group. Our results demonstrate that larval age can affect zebrafish locomotor behavior and should be considered when comparing light and dark responses across different studies. We also found that, at 5 dpf, there is no practical difference observed in locomotion between larvae who had undergone previous behavior testing and those who had not. This abstract does not reflect the official policy of the US EPA.
URLs/Downloads:
DOI: Effects of Larval Age and Previous Testing on Larval Zebrafish Behavior
NCSOT2023_POSTER_NBARBO.PDF (PDF, NA pp, 559.687 KB, about PDF)