Effects of 3,4-dichloroaniline on expression of ahr2 and cyp1a1 in zebrafish adults and embryos

Arylhydrocarbon receptor (Ahr) and cytochrome P4501a1 (Cyp1a1) are members of the Ahr/Cyp1a1 pathway that oxygenates various toxic chemicals including aryl hydrocarbons. To elucidate Ahr/Cyp1a1 pathway responses in teleost fish tissues, we examined the effects of 3,4-dichloroaniline (3,4-DCA), a reference toxic compound known to activate the Ahr/Cyp1a1 pathway, on the expression of arh and cyp1a1 in zebrafish tissues and embryos by means of in situ hybridization (ISH). Our ISH analysis showed that cyp1a1 expression was markedly activated by 3,4-DCA in the gill and intestinal epithelia, skin epidermis, and liver parenchymal cells of adult zebrafish. Before differentiation of the gill, intestine, and liver, skin was the site of cyp1a1 activation in embryos. Unlike the cyp1a1 response, 3,4-DCA-mediated ahr activation was not marked in either adults or embryos, indicating a possibility that stable ahr transcripts persist in the cytoplasm of these cells to induce cyp1a1. Young oocytes (previtellogenic to early vitellogic stage) express ahr; however activation of cyp1a1 by 3,4-DCA was negligible in these oocytes, suggesting that ahr expression in oocytes is not directly linked to cyp1a1 activation. Based on our finding that skin epidermis up-regulates cyp1a1 in response to 3,4-DCA, we demonstrated that fin explants, which can be harvested without sacrificing fish, can be used as a standard for assaying cyp1a1 activation in addition to embryos that are now used.