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Phenanthrene induces reproductive and developmental toxicity in fish, but whether it can disrupt the thyroid hormone balance and inhibit growth had not been determined to date. In this study, zebrafish embryos were exposed to phenanthrene (0, 0.1, 1, 10 and 100 μg/L) for 7 days. The results of this experiment demonstrated that phenanthrene induced thyroid disruption and growth inhibition in zebrafish larvae. Phenanthrene significantly decreased the concentration of l-thyroxine (T4) but increased that of 3,5,3'-l-triiodothyronine (T3). The expression of genes related to the hypothalamic-pituitary-thyroid (HPT) axis was altered in zebrafish larvae exposed to phenanthrene. Moreover, phenanthrene exposure significantly increased the malformation rate and significantly reduced the survival rate and the body length of zebrafish larvae. Furthermore, phenanthrene significantly decreased the concentrations of growth hormone (GH) and insulin-like growth factor-1 (IGF-1). Changes observed in gene expression patterns further support the hypothesis that these effects may be related to alterations along the GH/IGF-1 axis. In conclusion, our study indicated that exposure to phenanthrene at concentrations as low as 0.1 μg/L resulted in thyroid disruption and growth inhibition in zebrafish larvae. Therefore, the estimation of phenanthrene levels in the aquatic environment needs to be revisited., (Copyright © 2021. Published by Elsevier B.V.)

1,1-Trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethane (o,p'-DDT) and 1,1-dichloro-2,2-bis (p-chlorophenyl)-ethylene (p,p'-DDE) cause thyroid disruption, but the underlying mechanisms of these disturbances in fish remain unclear. To explore the potential mechanisms of thyroid dysfunction caused by o,p'-DDT and p,p'-DDE, thyroid hormone and gene expression levels in the hypothalamic-pituitary-thyroid (HPT) axis were measured, and the developmental toxicity were recorded in zebrafish larvae. Zebrafish embryos/larvae were exposed to o,p'-DDT (0, 0.28, 2.8, and 28 nM; or 0, 0.1, 1, and 10 μg/L) and p,p'-DDE (0, 1.57, 15.7, and 157 nM; or 0, 0.5, 5, and 50 μg/L) for 7 days. The genes related to thyroid hormone synthesis (crh, tshβ, tg, nis and tpo) and thyroid development (nkx2.1 and pax8) were up-regulated in both the o,p'-DDT and p,p'-DDE exposure groups. Zebrafish embryos/larvae exposed to o,p'-DDT showed significantly increased total whole-body T4 and T3 levels, with the expression of ugt1ab and dio3 being significantly down-regulated. However, the p,p'-DDE exposure groups showed significantly lowered whole-body total T4 and T3 levels, which were associated with up-regulation and down-regulation expression of the expression of dio2 and ugt1ab, respectively. Interestingly, the ratio of T3 to T4 was significantly decreased in the o,p'-DDT (28 nM) and p,p'-DDE (157 nM) exposure groups, suggesting an impairment of thyroid function. In addition, reduced survival rates and body lengths and increased malformation rates were recorded after treatment with either o,p'-DDT or p,p'-DDE. In summary, our study indicates that the disruption of thyroid states was different in response to o,p'-DDT and p,p'-DDE exposure in zebrafish larvae., (Copyright © 2019. Published by Elsevier B.V.)

Vigilin, a highly conserved protein from yeast to mammals, is a multifunctional protein in eukaryotic organisms. One biological function of vigilin is to stabilize the expression level of vitellogenin (VTG). This study aimed to develop vigilin as a new estrogen-inducible biomarker that correlates with the widely applied estrogen-inducible biomarker VTG and expand the ability to detect it in various species. Here, a recombinant monoclonal antibody with high specificity against the conserved C-terminal region of vigilin from zebrafish (Danio rerio) was successfully isolated from a phage display antibody library and found to recognize vigilin proteins from multiple vertebrate species. The effect of 17α-ethinylestradiol (EE2) on vigilin expression in the liver of zebrafish and juvenile crucian carp (Carassius auratus) was investigated. Although vigilin mRNA was expressed in all tissues analyzed from male zebrafish, vigilin protein was detected exclusively in the testis of male zebrafish, as well as the liver of female zebrafish and juvenile crucian carp at a lower level without exposure to EE2. Significant induction of vigilin mRNA by exposure to EE2 was observed in the liver and testis of male zebrafish, even at a low dose of 6.25 ng/L (21.09 pmol/L). In Hela cells, the expression of vigilin coincided with high protein synthesis activity but not dose-dependently by EE2 exposure. Therefore, the recombinant antibody may be used as a detection tool to screen for mammalian cell lines or organs with estrogen-inducible expression of vigilin., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Vitellogenin (VTG), the precursor of yolk proteins, is a sensitive biomarker of estrogenic contamination in aquatic environments. Traditionally, VTG was believed to be synthesized under the control of estrogen in the livers of mature females and then secreted into the blood, before being taken up by the ovaries and cleaved into lipovitellin and phosvitin, which provide nutrition for developing embryos. However, recent studies have reported that the liver is not the only tissue to express VTG and this has led to questions over the precise tissue distribution of VTG in zebrafish. Moreover, studies in zebrafish on the expression of the VTG protein are rare. Using Western blotting and reverse-transcriptase polymerase chain reaction, this present study reports that the VTG protein and VTG1 mRNA were detected not only in the liver, but also in various extrahepatic tissues, including the heart, spleen, kidney, skin, muscle, gill, eye and brain tissues, of female and 17α-ethinylestradiol (EE2)-treated male zebrafish. Due to the high expression levels of VTG and the ease of taking samples, skin and eye tissues were chosen to evaluate the effects of varying doses and exposure times of EE2 on male zebrafish. The VTG gene and protein were induced by EE2 exposure in liver, skin and eye tissues of male zebrafish in dose- and time-dependent patterns. Therefore, we suggest that zebrafish skin and eye tissues may be alternatives to plasma and liver tissues for VTG biomarker analysis., (Copyright © 2014 Elsevier B.V. All rights reserved.)