Estrogen-Responsive Transient Expression Assay Using a Brain Aromatase-Based Reporter Gene in Zebrafish (Danio rerio)

摘要

Whereasendogenousestrogensplayanimportantroleinthedevelopment,maintenance,andfunctionoffemaleandmalereproductiveorgans,xenoestrogenspresentintheenvironmentdisruptnormalendocrinefunctioninhumansandwildlife.Variousinvivoandinvitroassayshavebeendevelopedtoscreenthesexenoestrogens.However,traditionalinvivoassaysarelaboriousandunsuitableforlarge-scalescreening,andinvitroassaysdonotnecessarilyreplicateinvivofunctioning.Toovercometheselimitations,wedevelopedatransientexpressionassayinzebrafish,intowhichabrainaromatase(cyp19a1b)-basedestrogen-responsivereportergenewasintroduced.Inresponseto17#59137;-estradiol(10supxmlns="http://pub2web.metastore.ingenta.coms/"#8722;6/supM)andheptachlor(10supxmlns="http://pub2web.metastore.ingenta.coms/"#8722;6/supM),zebrafishembryoscarryingthereporterconstructexpressedenhancedgreenfluorescentproteinintheolfactorybulb,telencephalon,preopticarea,andmediobasalhypothalamus.Thissystemwillservetomodeltheinvivoconversionandbreakdownofestrogeniccompoundsandthusprovidearapidpreliminaryscreeningmethodtoestimatetheirestrogenicity.