title: Modulation of steroidogenesis and estrogen signalling in the estuarine killifish (Fundulus heteroclitus) exposed to ethinylestradiol.
abstract: Previous studies have shown that mummichog (Fundulus heteroclitus; a lunar, asynchronous-spawning killifish of the western Atlantic) exposed to 17alpha-ethynylestradiol (EE2) exhibit decreased plasma reproductive steroid levels, decreased gonadal steroid production, increased plasma vitellogenin, decreased fecundity and impaired fertilization. The objective of this study was to determine the potential mechanisms by which EE2 depresses gonadal steroidogenesis and influences estrogen signalling in the mummichog. Adult recrudesced fish were exposed to the potent synthetic estrogen, ethinylestradiol (EE2; 0-270ng/L) for 14 days. Following exposure, gonadal tissue was removed and incubated for 24h with stimulators of steroidogenesis, including forskolin; 25-OH cholesterol; or pregnenolone. Testosterone production was decreased in basal, forskolin-stimulated and pregnenolone-stimulated EE2-exposed males, indicating effects on the steroidogenic pathway both at and downstream of cholesterol mobilization to P450 side-chain cleavage (P450scc) and/or P450scc conversion of cholesterol to pregnenolone. Hepatic transcript levels of estrogen receptor alpha (ERalpha) and vitellogenin were increased in EE2-treated males compared to control recrudescing males and females confirming an estrogenic response. Hepatic heat shock protein 90 (Hsp90), a chaperoning molecule involved in estrogen signalling, was not affected by EE2 exposure at either the transcript or protein level. However, higher levels of Hsp90 observed in the membrane fractions of female fish raise interesting questions regarding the influence of gender on Hsp90's role in estrogen signalling. These results demonstrate that EE2 can alter steroid production at specific sites within the steroidogenic pathway and can stimulate hepatic estrogen signalling, providing important information regarding the molecular mechanisms underlying the endocrine response of the mummichog to exogenous estrogen.
authors.affiliation: Department of Biology and Canadian Rivers Institute, University of Prince Edward Island, Charlottetown, PE, Canada, C1A 4P3. nshogan@upei.ca <nshogan@upei.ca>
authors.forename: Natacha S
authors.initials: NS
authors.lastname: Hogan
authors.forename: Suzanne
authors.initials: S
authors.lastname: Currie
authors.forename: Sacha
authors.initials: S
authors.lastname: LeBlanc
authors.forename: L Mark
authors.initials: LM
authors.lastname: Hewitt
authors.forename: Deborah L
authors.initials: DL
authors.lastname: MacLatchy
mesh_terms: D000818:Animals; D005576:Colforsin / Q000633:toxicity; D003577:Cytochrome P-450 Enzyme System / Q000097:blood / Q000378:metabolism; D052244:Endocrine Disruptors / Q000097:blood / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000097:blood / Q000378:metabolism; D004967:Estrogens / Q000097:blood / Q000378:metabolism*; D004997:Ethinyl Estradiol / Q000633:toxicity*; D005260:Female; D005306:Fertilization / Q000187:drug effects / Q000502:physiology; D023781:Fundulidae / Q000235:genetics / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000097:blood / Q000378:metabolism; D006888:Hydroxycholesterols / Q000633:toxicity; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D011284:Pregnenolone / Q000633:toxicity; D015398:Signal Transduction / Q000187:drug effects; D013256:Steroids / Q000096:biosynthesis*; D013739:Testosterone / Q000097:blood / Q000378:metabolism; D013997:Time Factors; D014819:Vitellogenins / Q000097:blood / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D052244:Endocrine Disruptors; D047628:Estrogen Receptor alpha; D004967:Estrogens; D006360:Heat-Shock Proteins; D006888:Hydroxycholesterols; D013256:Steroids; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D005576:Colforsin; D013739:Testosterone; D004997:Ethinyl Estradiol; D011284:Pregnenolone; D003577:Cytochrome P-450 Enzyme System