title: Relationship between brain and ovary aromatase activity and isoform-specific aromatase mRNA expression in the fathead minnow (Pimephales promelas).
abstract: There is growing evidence that some chemicals present in the environment have the capacity to inhibit, or potentially induce, aromatase activity. This study compared aromatase activities and isoform-specific mRNA expression in brain and ovary tissue from non-exposed fathead minnows representing three different ages and stages of reproductive activity, and from fathead minnows exposed to the aromatase inhibitor fadrozole for 7d. The goal was to determine whether measures of a single aromatase endpoint in either brain or ovary tissue would be sufficient to understand and predict system-wide effects of endocrine disrupting chemicals on aromatase activity and transcript levels. Aromatase activity in the ovary, but not brain, varied significantly with age/reproductive category, with adults held in non-reproductive conditions showing significantly lower activity than juveniles and reproductively-active adults. Significant correlations between isoform-specific transcript levels and aromatase activity were observed for ovary tissue, but those relationships were not robust for all age/reproductive categories, nor were they sustained in fadrozole-treated fish. In vitro, fadrozole inhibited the aromatase activity of brain and ovary post-mitochondrial supernatants with similar potency (IC50s = 8.82 +/- 1.58 and 6.93 +/- 0.80 microM for brain and ovary, respectively), despite large differences in the magnitude of activity. In vivo, fadrozole altered aromatase activity and isoform-specific transcript levels in both brain and ovary tissue, but concentration-response relationships were different for each tissue. Aromatase activity and P450aromB mRNA expression in brain showed a dose-dependent decrease at concentrations greater than 5.55 microg/L. In contrast, ovary activity showed an inverted U-shaped concentration-response consistent with the interplay between increased P450aromA transcript levels in ovary and competitive inhibition of the aromatase enzyme. As a whole, results of this study did not reveal any robust correlations between brain and ovary aromatase activity and/or isoform-specific mRNA expression. However, they were consistent with the current body of evidence related to teleost aromatase regulation, suggesting that increased understanding of the biology of aromatase may facilitate system-wide understanding of effects on aromatase based on relatively few measured endpoints.
authors.affiliation: US Environmental Protection Agency, Mid-Continent Ecology Division, Duluth, MN, USA. villeneuve.dan@epa.gov
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Villeneuve
authors.forename: Iris
authors.initials: I
authors.lastname: Knoebl
authors.forename: Michael D
authors.initials: MD
authors.lastname: Kahl
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Jensen
authors.forename: Dean E
authors.initials: DE
authors.lastname: Hammermeister
authors.forename: Katie J
authors.initials: KJ
authors.lastname: Greene
authors.forename: Lindsey S
authors.initials: LS
authors.lastname: Blake
authors.forename: Gerald T
authors.initials: GT
authors.lastname: Ankley
mesh_terms: D000367:Age Factors; D000818:Animals; D001141:Aromatase / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D047072:Aromatase Inhibitors / Q000633:toxicity; D001921:Brain / Q000187:drug effects / Q000201:enzymology*; D003530:Cyprinidae / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D017316:Fadrozole / Q000633:toxicity; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D008297:Male; D010053:Ovary / Q000187:drug effects / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D020033:Protein Isoforms / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D047072:Aromatase Inhibitors; D017931:DNA Primers; D020033:Protein Isoforms; D012333:RNA, Messenger; D001141:Aromatase; D017316:Fadrozole