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Traditional vs. Novel: Using Alternative Analysis Methods to Identify Negative Control Chemicals in Larval Zebrafish Behavior Assays (TSN 2024)
Citation:
Knapp, B., D. Hunter, M. Lowery, J. Olin, K. Jarema, Z. Rowson, AND S. Padilla. Traditional vs. Novel: Using Alternative Analysis Methods to Identify Negative Control Chemicals in Larval Zebrafish Behavior Assays (TSN 2024). Presented at 9th Annual Triangle Society for Neuroscience (TSfN), Durham, NC, April 26, 2024. https://doi.org/10.23645/epacomptox.25872679
Impact/Purpose:
Poster presented to 9th Annual Triangle Society for Neuroscience
Description:
Identifying reliable positive and negative control compounds is integral to determining the sensitivity and specificity of chemical assays. Our laboratory screens chemicals for developmental neurotoxicity potential using assays that assess larval zebrafish locomotor behavior in response to visual stimuli. For such assays, there are established positive controls; however, well-characterized negative controls are lacking. Using a set of nine chemicals that appeared to have no evidence of developmental neurotoxicity in mammals (PMID: 35908584), we performed a range-finding study to assess lethality and teratogenic potential of each chemical (≤ 100µM). Next, using non-toxic concentrations, the locomotor activity profile was assessed at 6 days post fertilization using a light/dark transition assay. The data was analyzed using two methods: (1) the traditional method compared the average distance moved in the light period to the dark period using a repeated measures ANOVA and (2) a novel approach analyzed various aspects of the locomotor profile (e.g., average speed, habituation, maximum and minimum activity, aspects of the startle response, range of activity, and area under the curve [AUC]) and constructed a benchmark concentration for each if there was a significant concentration-related change. The traditional method showed that none of the nine chemicals tested (i.e., L-ascorbic acid, D-mannitol, saccharin, sodium benzoate, metformin hydrochloride, ibuprofen, glycerol, omeprazole, and selegiline hydrochloride) had an effect. However, the novel method uncovered a significant change in average speed in the light for selegiline hydrochloride. Therefore, the novel approach may be more sensitive to behavioral changes than the traditional method. Standardizing a set of negative control chemicals is essential for researchers to properly evaluate the behavioral results obtained from unknown chemicals and will assist with assessing new chemical screening methods. This abstract does not reflect the official policy of the US EPA.
URLs/Downloads:
DOI: Traditional vs. Novel: Using Alternative Analysis Methods to Identify Negative Control Chemicals in Larval Zebrafish Behavior Assays (TSN 2024)
4.24.2024 TRIANGLE SFNCHAPTER 2024 POSTER.PDF (PDF, NA pp, 1223.915 KB, about PDF)