Lipidomics reveals multiple stressor effects (temperature × mitochondrial toxicant) in the zebrafish embryo toxicity test.

Aquatic organisms are exposed to multiple stressors in the environment, including contaminants and rising temperatures due to climate change. The objective of this study was to characterize the effect of increased temperature on chemical-induced toxicity and lipid profiles during embryonic development and hatch in fish. This is important because temperature and many environmental chemicals modulate cellular metabolism and lipids, both of which play integral roles for normal embryonic development. As such, we employed the zebrafish embryo toxicity test for multiple stressor exposures, using the mitochondrial toxicant 2,4-Dinitrophenol (DNP; 6-30 μM) in conjunction with different temperature treatments (28 °C and 33 °C). We found a positive relationship between temperature and lethality at lower DNP concentrations, suggesting temperature stress can increase toxicant sensitivity. Next, we used LC-MS/MS for lipidomics following exposure to sublethal stressor combinations. It was determined that temperature stress at 33 °C augmented DNP-induced effects on the lipidome, including the upregulation of bioactive lipids involved in apoptosis (e.g., ceramides). These data reveal potential implications for climate change and sensitivity to environmental pollution and demonstrate the utility of lipidomics to characterize metabolic pathways underlying toxicity. Data such as these are expected to advance adverse outcome pathways by establishing multiple stressor networks that include intermediate lipid responses.