Neurotoxicity induced by difenoconazole in zebrafish larvae via activating oxidative stress and the protective role of resveratrol

Difenoconazole (DIF) is a typical triazole fungicide detected in the aquatic ecosystem and organisms. However, the neurotoxic effects of DIF remain largely unknown. This study aimed to investigate the neurotoxicity of DIF in zebrafish and the underlying neuroprotective properties of resveratrol (RES, an antioxidant polyphenol). Zebrafish embryos/larvae were treated with 0.6 and 1.2 mg/L DIF from 4 to 96 h post fertilization (hpf) and neurodevelopment was systematically assessed. DIF induced developmental toxicity and aberrant neurobehaviors, including decreased movement time, swimming distance and clockwise rotation times. DIF suppressed the neurogenesis of the central nervous system (CNS) in HuC:egfp transgenic zebrafish and the length of motor neuron axon in hb9:egfp transgenic zebrafish. DIF inhibited cholinesterase activities and downregulated neurodevelopment related genes. DIF also increased oxidative stress via excessive production of reactive oxygen species and decreased activities of antioxidant enzymes, subsequently triggering neuronal apoptosis in the brain. RES partially reinstated DIF-induced neurotoxicity and developmental toxicity by inhibiting excessive oxidative stress and apoptosis, suggesting the involvement of oxidative stress in DIF-induced neurotoxicity. Overall, this study identified the potential mechanisms underlying DIF-induced neurotoxicity and suggested RES as a promising therapeutic strategy.