The impaired swim bladder via ROS-mediated inhibition of the Wnt / Hedgehog pathway in zebrafish embryos exposed to eight toxic chemicals and binary chemical mixtures

Abstract

To comprehensively explore the potential toxicity of aquatic organisms exposed to chlorinated or brominated flame retardants (BFRs) and metals mixtures, it is necessary to find a common pathway to relate local toxic targeted sites or organs. A key challenge in environmental risk assessment (ERA) is how to clarify the same or different sites or organs of toxic action in a species after exposure to individual chemicals or chemical mixtures. In this study, zebrafish embryo was used to evaluate the sub-lethal toxicity (swim bladder damage) of tris(2,3-dibromo propyl) isocyanurate (TBC), chlorinated paraffins (CPs), hexabromocyclododecane (HBCD), Cu, Cd, Pb, Ag, and Zn through optical microscopy methods, and corresponding sub-lethal molecular levels (inflammation-related enzymes [deiodinase (DIO) enzymes] and transcriptional levels of key genes) in fish through quantitative real-time PCR (qRT-PCR). The tested chemicals all caused failed inflation of the swim bladder, as indicated by activity inhibition of type 2 iodothyronine deiodinase enzyme. Following embryonic exposure to respective TBC + Cu, HBCD + TBC, and Cd + Pb mixtures, as the concentration of the respective Cu, TBC, and Pb increased, the deformity of the swim bladder increased, as also indicated by activity inhibition of type 2 iodothyronine deiodinase enzyme. Additionally, eight chemicals down-regulated Wnt (wnt3, wnt9b, fzd3b, wnt1, fzd5, and fdz1) signaling pathways, which were neurotoxic responses to individual chemical treatments and Hedgehog (ihh, shh, ptc1 and ptc2) signaling pathways. Moreover, excessive ROS induced by eight chemicals effectively induced defects in the swim bladder and Wnt/Hedgehog signaling, which also be proved in respective TBC + Cu, HBCD + TBC, and Cd + Pb mixture treatments. Our results first revealed that eight chemicals caused swim bladder developmental defects via ROS-mediated inhibition of the Wnt and Hedgehog pathways, which revealed the common targeted sites or organs (swim bladders) for further studying the toxic mechanisms underlying the chemical mixtures.