title: Effects of Fusarium mycotoxins on steroid production by porcine granulosa cells.
abstract: Fusarium mycotoxins, such as trichothecenes and zearalenone, are common grain and foodstuffs contaminants. Some of these like deoxynivalenol (DON) can negatively impact pregnancy success in swine, but evidence for direct ovarian effects of DON, zearalenone, and its major metabolite, alpha-zearalenol (ZEA) is meager. To evaluate the effects of two mycotoxins, DON and ZEA on porcine granulosa cell(s) (GC) proliferation, steroidogenesis and gene expression, pig GC from small follicles (1-5mm) were cultured for 2 days in 5% fetal bovine serum and 5% porcine serum-containing medium followed by 2 days in serum-free medium containing control (no mycotoxins) or mycotoxins (at various doses/combinations). Both DON and ZEA had biphasic effects on IGF-I-induced estradiol production, increasing estradiol production at smaller doses and inhibiting at larger doses. ZEA at 3,000 ng/mL (9.37 microM) increased IGF-I-induced progesterone production and at 30 ng/mL (0.0937 microM) and 300 ng/mL (0.937 microM) were without effect, but these doses of ZEA increased FSH-induced progesterone production. ZEA at 3,000 ng/mL inhibited FSH plus IGF-I-induced CYP19A1 and CYP11A1 mRNA abundance. DON inhibited progesterone production at 100 ng/mL (0.337 microM) and 1,000 ng/mL (3.37 microM) but at 10 ng/mL (0.0337 microM) was without effect. DON at 1,000 ng/mL (but not at 10 ng/mL) completely inhibited FSH plus IGF-I-induced CYP19A1 and CYP11A1 mRNA abundance. The concomitant treatment of ZEA had little effect on the dose response to DON. DON increased IGF-I-induced cell numbers at 10 and 100 ng/mL and inhibited cell numbers at 1,000 ng/mL, whereas ZEA had no effect on GC numbers. Only a combined treatment of DON and ZEA increased serum-induced cell proliferation. In conclusion, mycotoxins have direct dose-dependent effects on GC proliferation, steroidogenesis and gene expression. These direct ovarian effects could be one mechanism whereby contaminating Fusarium mycotoxins in feedstuffs could impact reproductive performance in swine.
authors.affiliation: Department of Veterinary Clinical Science, University of Milan, via Celoria, Milan, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Ranzenigo
authors.forename: Francesca
authors.initials: F
authors.lastname: Caloni
authors.forename: Fausto
authors.initials: F
authors.lastname: Cremonesi
authors.forename: Pauline Y
authors.initials: PY
authors.lastname: Aad
authors.forename: Leon J
authors.initials: LJ
authors.lastname: Spicer
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics / Q000378:metabolism; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone; D005670:Fusarium / Q000378:metabolism*; D005786:Gene Expression Regulation; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis*; D013552:Swine; D014255:Trichothecenes / Q000494:pharmacology*; D015029:Zeranol / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D012333:RNA, Messenger; D013256:Steroids; D014255:Trichothecenes; C029659:zearalenol; D007334:Insulin-Like Growth Factor I; D015029:Zeranol; D005640:Follicle Stimulating Hormone; D019363:Cytochrome P-450 CYP1A1; D002786:Cholesterol Side-Chain Cleavage Enzyme; C007262:deoxynivalenol