Abstract
Estrogenic contaminants released into water bodies are potentially affecting the reproduction of aquatic organisms. Exposure to 17α-ethinylestradiol (EE2), a synthetic estrogen agonist commonly found in sewage effluents, has been shown to cause gonadal changes in male gonochoristic fish ranging from gonadal intersex to complete sex reversal. Although these gonadal changes have been well studied in Japanese medaka Oryzias latipes, the molecular mechanisms behind them are poorly understood. Our objective was to study the signaling pathways elicited by exposure to different concentrations of EE2 in this species. Embryos and larvae were sexed by the presence of leucophores and dmy expression (only in males). Male medaka were exposed to two EE2 concentrations (30 and 300 ng/L) during their gonadal differentiation period (7–22 dpf). The transcriptome of larvae was analyzed using RNA sequencing followed by pathway analysis. Genes involved in sex differentiation and gonadal development (e.g., cldn19, ctbp1, hsd17b4) showed a female-like expression pattern in EE2-exposed males with some genes changing in expression in an EE2 concentration-dependent manner. However, not all genes known to be involved in sex differentiation and gonadal development (e.g., wnt4b) were altered by EE2. Several of the prominently affected signaling pathways involved genes associated with steroidogenesis, steroid receptor signaling and steroid metabolism, such as cyp2b3, cyp3b40, cyp1a, hsd17b4. We also report on novel genes and pathways affected that might play a role in gonadal changes, including several genes associated with FXR/RXR and LXR/RXR activation networks. This study is the first to examine the transcriptomic changes in male fish resulting from exposure to EE2 during the gonadal differentiation period, providing new insights on the signaling pathways involved in the development of gonadal changes in gonochoristic fish.
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