A Novel Organophosphate Ester, Tris(2,4-di<i>tert</i>-butylphenyl) Phosphate, Induced Reproductive Toxicity in Male Zebrafish at Environmentally Relevant Concentrations

As a novel organophosphate ester (NOPE), tris(2,4-di<i>tert</i>-butylphenyl) phosphate (TDtBPP) has attracted significant attention due to its unexpectedly high detection in natural environments. However, the ecological toxic effects of environmentally relevant concentrations of TDtBPP in organisms remain entirely unknown. In this study, 1 month old zebrafish were exposed to 0, 50, 500, or 5000 ng/L TDtBPP for 150 days, and the reproductive toxicity in male fish was evaluated. Results demonstrated that TDtBPP exposure significantly inhibited the maturation of spermatozoa and thus decreased spermatogenesis. Furthermore, abnormal sperm morphology and decreased sperm motility were also observed. The decrease in sperm quantity and quality eventually resulted in the declining fecundity. Moreover, TDtBPP exposure downregulated the expression of <i>hsd3b1 in vivo</i> and in <i>vitro</i> and subsequently inhibited the synthesis of androgens in zebrafish testes and Leydig cells. This inhibition of androgen synthesis appeared to be responsible for the observed reproductive toxicity in male fish. Molecular docking and dual-luciferase reporter gene experiments elucidated that TDtBPP inhibited the promotion of vitamin D on <i>hsd3b1</i> transcription by the vitamin D receptor and thus downregulated the expression of <i>hsd3b1</i>. Our findings provide first time evidence that TDtBPP poses a risk to male fish reproduction at environmentally relevant levels.