title: Effects of low concentrations of the antiprogestin mifepristone (RU486) in adults and embryos of zebrafish (Danio rerio): 2. Gene expression analysis and in vitro activity.
abstract: Here, we analyzed the transcriptional effects of the antiprogestin mifepristone (MIF, RU486) and progesterone (P4) in zebrafish as well as their in vitro activities in yeast-based reporter gene assays. This study is associated with the reproduction study in adult zebrafish and embryos exposed for 21 days to 5, 39, 77 ng/L MIF, and 25 ng/L P4 (Blüthgen et al., 2013a). The in vitro activities of MIF and P4 were investigated using a series of recombinant yeast-based assays (YES, YAS, YPS) and compared to transcriptional alterations obtained in fish tissues and embryos from the exposure study. MIF elicited antiestrogenic, androgenic and progestogenic activities in recombinant yeast, similar to P4, and no antiprogestogenic activity in vitro. The transcriptional alterations of steroid hormone receptors were similar in adult males and females, and more pronounced in embryos. MIF tended to transcriptionally down-regulate the androgen (ar), progesterone (pgr) and glucocorticoid (gr) receptors in adult fish and embryos. Transcripts of the estrogen receptor (esr1) and vitellogenin (vtg1) were not significantly altered. A trend for down-regulation was observed for transcripts of genes belonging to steroidogenic enzymes including 17β-hydroxysteroid dehydrogenase type 3 (hsd17b3), 3 β-hydroxysteroid dehydrogenase (hsd3b), P450 aromatase A (cyp19a) and 11β-hydroxylase (cyp11b). P4 resulted in similar transcriptional alterations as MIF. The data indicate that gene expression changes (here and later gene expression is taken as synonym to gene transcription) and in vitro activities match only in part including the lack of antiprogestogenic activity of MIF. Additionally, effects on reproduction and gonad histology described in the associated report (Blüthgen et al., 2013a) can only partly be explained by gene expression data presented here.
authors.affiliation: University of Applied Sciences Northwestern Switzerland, School of Life Sciences, Institute for Ecopreneurship, Gründenstrasse 40, 4132 Muttenz, Switzerland; University of Basel, Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, Klingelbergstrasse 50, 4056 Basel, Switzerland.
authors.forename: Nancy
authors.initials: N
authors.lastname: Blüthgen
authors.forename: John P
authors.initials: JP
authors.lastname: Sumpter
authors.forename: Alex
authors.initials: A
authors.lastname: Odermatt
authors.forename: Karl
authors.initials: K
authors.lastname: Fent
mesh_terms: D000818:Animals; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D008297:Male; D015735:Mifepristone / Q000633:toxicity*; D014819:Vitellogenins / Q000235:genetics / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000196:embryology / Q000502:physiology*
chemical_list: D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D015735:Mifepristone
keywords: Antiprogestin; Gene expression; In vitro hormonal activity; Mifepristone; Progesterone; RU486; Recombinant yeast system