Zebrafish Embryonic Exposures and Imaging

Procedures were performed in accordance with the Icahn School of Medicine at Mount Sinai Institutional Animal Care and Use Committee (IACUC). Adult wild-type (WT; AB, Tab14 and TAB5) and Tg(fabp10a:nls-mCherry^mss4Tg) (Mudbhary et al., 2014 (link)) fish were maintained on a 14:10 light:dark cycle at 28°C. Fertilized embryos from natural spawning of group matings were collected and cultured in embryo water (0.6 g/l Crystal Sea Marinemix; Marine Enterprises International, Baltimore, MD) containing Methylene Blue at 28°C. Embryos were treated with sodium (meta)arsenite (Sigma, S7400) beginning at 4 h post fertilization (hpf). sodium (meta)arsenite and/or ethanol were diluted from stock solutions in 10 ml of embryo water. After addition of exposure medium, 35-mm dishes were sealed with Parafilm and returned to the incubator. Medium was not replaced during the exposure period unless otherwise noted. For co-exposures, sodium (meta)arsenite was removed and replaced with embryo water containing sodium (meta)arsenite and tunicamycin at 72 hpf or sodium (meta)arsenite and ethanol at 96 hpf at the indicated concentrations. Images of anesthetized larvae were taken by mounting in 3% methyl cellulose using a Nikon SMZ1500 stereomicroscope.