Long-term effects of individual and combined exposure to microplastics and copper in zebrafish hypothalamic-pituitary-gonadal axis - A multi-biomarker evaluation.

Microplastics (MPs) pollution and metal contamination are two prominent environmental stressors with multifaceted implications for aquatic life and ecosystem health. However, the underlying toxicological mechanisms of MPs and metals co-exposure on fish reproduction processes are largely unknown. In this study, zebrafish (Danio rerio) were exposed to MPs (2 mg/L), copper (25 μg/L, Cu25), and their mixture (Cu25 + MPs), for 30 days. The oxidative stress response, along with the expression profile of the hypothalamic-pituitary-gonadal (HPG) axis-related genes in the brain and gonad of zebrafish, were evaluated. The findings demonstrated that exposure to MPs and Cu affects the antioxidant system of zebrafish brain and gonads, inhibiting GPx in individuals exposed to MPs, Cu25, and their mixture. The gene expression analysis revealed dysregulation of the HPG axis-related genes. Specifically, the androgen receptor (ar), estrogen receptor 1 (esr1), follicle-stimulating hormone (fsh), and gonadotropin-releasing hormone 2 (gnrh2) were upregulated in the brain, whereas the genes esr2a, ar, cytochrome P450 family 11 subfamily A member 1 (cyp11a) and cyp19a were upregulated in the gonads. Both the biochemical and gene expression results showed that the brain and gonads were differently affected by MPs and Cu with the effects varying with fish gender. Furthermore, the mixture exposure affected the brain the most, and the individual pollutants affected the gonads the most. Overall, this study highlights that MPs, alone or combined with Cu, adversely affect the HPG axis of zebrafish, posing a potential threat to the reproduction of fish populations.