Metabolomics reveals the reproductive abnormality in female zebrafish exposed to environmentally relevant levels of climbazole

Highlights: • Exposure to real environmental levels of CBZ inhibited the fecundity of zebrafish. • Metabolomics and biochemical analysis were used to elucidate the action mechanism. • Altered steroidogenesis can play a vital role in CBZ-induced reproductive toxicity. • CBZ induced oxidative stress and apoptosis in zebrafish ovaries. • Mass spectrometry imaging was introduced to confirm the metabolomics results. Climbazole (CBZ) ubiquitously detected in the aquatic environment may disrupt fish reproductive function. Thus far, the previous study has focused on its transcriptional impact of steroidogenesis-related genes on zebrafish, but the underlying toxic mechanism still needs further investigation at the metabolic level. In this study, adult zebrafish were chronically exposed to CBZ at concentrations of 0.1 (corresponding to the real concentration in surface water), 10, and 1000 μg/L and evaluated for reproductive function by egg production, with subsequent ovarian tissue samples taken for histology, metabolomics, and other biochemical analysis. After 28 days' exposure, fecundity was significantly decreased in all exposure groups, with the inhibition of oocytes in varying developmental stages to a certain degree. The decrease in retinoic acid and sex hormones, down-regulated genes important in steroidogenesis, and increase in oxidized/reduced glutathione ratio and occurrence of apoptotic cells were observed in zebrafish ovaries following exposure to CBZ even at environmentally realistic concentrations, suggesting that alternations in steroidogenesis and oxidative stress can play significant roles in CBZ-triggered reproductive toxicity. Besides, mass spectrometry imaging analysis validated the results from metabolomics analysis. Our findings provide novel perspectives for unveiling the mechanism of reproductive dysfunction by CBZ and highlight its risk to fish reproduction.