Abstract
Dimethyl sulfoxide (DMSO) is a popular solvent for developmental toxicity testing of chemicals and pharmaceuticals in zebrafish embryos. In general, it is recommended to keep the final DMSO concentration as low as possible for zebrafish embryos, preferably not exceeding 100 μL/L (0.01%). However, higher concentrations of DMSO are often required to dissolve compounds in an aqueous medium. The aim of this study was to determine the highest concentration of DMSO that can be safely used in our standardized Zebrafish Embryo Developmental Toxicity Assay (ZEDTA). In the first part of this study, zebrafish embryos were exposed to different concentrations (0–2%) of DMSO. No increase in lethality or malformations was observed when using DMSO concentrations up to 1%. In a follow-up experiment, we assessed whether compounds that cause no developmental toxicity in the ZEDTA remain negative when dissolved in 1% DMSO, as false positive results due to physiological disturbances by DMSO should be avoided. To this end, zebrafish embryos were exposed to ascorbic acid and hydrochlorothiazide dissolved in 1% DMSO. Negative control groups were also included. No significant increase in malformations or lethality was observed in any of the groups. In conclusion, DMSO concentrations up to 1% can be safely used to dissolve compounds in the ZEDTA.
Highlights
Zebrafish embryos are gaining interest as an alternative to animal testing for developmental toxicity screening of candidate drugs and chemicals
Zebrafish embryo-based assays are already used for this purpose by different research groups (Brannen et al, 2010; Van den Bulck et al, 2011; Gustafson et al, 2012; Pruvot et al, 2012; Selderslaghs et al, 2012; Lee et al, 2013; Teixidó et al, 2013; Ball et al, 2014; Yamashita et al, 2014; Song et al, 2021) and in our group we refer to this assay as the ZEDTA, i.e. Zebrafish Embryo Developmental Toxicity Assay (Pype et al, 2015; Hoyberghs et al, 2020; Bars et al, 2021)
The 2% Dimethyl sulfoxide (DMSO) group has more than 10% malformed and/or dead larvae, which means that using 2% DMSO as a solvent control group makes the experiment invalid, and cannot be used
Summary
Zebrafish embryos are gaining interest as an alternative to animal testing for developmental toxicity screening of candidate drugs and chemicals. Zebrafish embryo-based assays are already used for this purpose by different research groups (Brannen et al, 2010; Van den Bulck et al, 2011; Gustafson et al, 2012; Pruvot et al, 2012; Selderslaghs et al, 2012; Lee et al, 2013; Teixidó et al, 2013; Ball et al, 2014; Yamashita et al, 2014; Song et al, 2021) and in our group we refer to this assay as the ZEDTA, i.e. Zebrafish Embryo Developmental Toxicity Assay (Pype et al, 2015; Hoyberghs et al, 2020; Bars et al, 2021). Many xenobiotics are rather hydrophobic (Weigt et al, 2011; Ball et al, 2014), and organic solvents are needed to solubilize the compounds of interest for exposure experiments in zebrafish embryos (Hutchinson et al, 2006)
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