Reproductive toxicity and molecular mechanisms of benzo[a]pyrene exposure on ovary, testis, and brood pouch of sex-role-reversed seahorses (Hippocampus erectus)

Abstract

As a flagship taxa for marine ecosystems, seahorses possess a unique reproductive strategy of “male pregnancy”. They are severely threatened by marine petroleum-based pollution but the molecular mechanism involved remains unclear. We evaluated the toxic effects and mechanisms of sub-acute exposure to benzo[a]pyrene (BaP), a representative polycyclic aromatic hydrocarbon (PAH), at three environmental related dosages (0.5, 5, and 50 μg/L) on the reproductive organs of sex-role-reversed lined seahorses (Hippocampus erectus). BaP induced ovary, testis and brood pouch tissue damage in a concentration-dependent manner. Transcriptomic results suggested that reproductive organs of the seahorse could undergo biotransformation and detoxification of BaP via the P450 enzyme system; the differential expression of key genes related to these pathways determined the differences in the accumulation of toxic substances in the reproductive organs. Moreover, toxic substances directly induced differential tissue damage in situ by activating tissue-specific signaling pathways: BaP-induced ovarian apoptosis and failure by upregulation of the pro-apoptotic genes, vadc1, traf2b, tnfsf10, and pycard (P < 0.05); inhibition of testicular function through disruption of genes associated with “ECM-receptor interaction”, “Tight junction”, and “Spermatogenesis” pathways; and interference with brood pouch immune responses, significantly suppressing the expression of ripk1, il-1b, casp3a, apaf1, calr, and canx (P < 0.05), thereby impairing “Apoptosis”, “Phagosome”, and “Necroptosis” processes, ultimately compromising brood pouch maintenance. Toxic substances exacerbate damage to the reproductive organs in seahorses by disrupting Ca²⁺ homeostasis. At environmentally-relevant concentrations of BaP, the reproductive efficiency of seahorses may be severely affected, increasing the risk of a decline in the abundance and diversity of wild populations.