Pyriproxyfen (PPF), a broad-spectrum insecticide known to cause reproductive and endocrine disruption in invertebrates, while the data is scarce in aquatic vertebrates. The goal of this study is to investigate the impact of PPF on reproductive endocrine system of male and female zebrafish along hypothalamus-pituitary-gonadal (HPG) axis. In brain, PPF caused significant alteration in the transcripts of erα, lhβ, and cyp19b genes in male and fshβ, lhβ, and cyp19b genes in female zebrafish. The downstream genes of steroidogenic pathway like, star, 3βhsd, 17βhsd, and cyp19a expression were significantly altered in gonad of both sexes. Subsequent changes in circulatory steroid hormone levels lead to imbalance in hormone homeostasis as revealed from estradiol/testosterone (E2/T) ratio. Further, the vitellogenin transcript level was enhanced in hepatic tissues and their blood plasma content was increased in male (16.21%) and declined in female (21.69%). PPF also induced histopathological changes in gonads such as, reduction of mature spermatocytes in male and vitellogenic oocytes in female zebrafish. The altered E2/T ratio and gonadal histopathology were supported by the altered transcript levels of HPG axis genes. Overall, these findings provide new insights of PPF in zebrafish reproductive system and highlights for further investigations on its potential risks in aquatic environment. Display omitted •The endocrine disruptive potential of pyriproxyfen (PPF) was studied in zebrafish.•PPF altered the gonadotropin and steroidogenic transcript levels in both sexes.•Blood plasma concentration of testosterone and estradiol were significantly differed.•Vitellogenin expression was enhanced in liver and varied in blood plasma level.•PPF exposure caused histopathological damages in testes and ovary of zebrafish.