Sex-biased microRNAs(miRNAs)influence gonadal development in fish by directly targeting genes associated with estrogen production pathways.WW domain-binding protein 2(WBP2)functions as a crucial transcriptional co-act...Sex-biased microRNAs(miRNAs)influence gonadal development in fish by directly targeting genes associated with estrogen production pathways.WW domain-binding protein 2(WBP2)functions as a crucial transcriptional co-activator of the estrogen and progesterone receptors(PGR).This study investigates the direct modulation of a sex-biased miR-133b on wbp2 and its regulatory role in gonadal development in fish,the greater amberjack(Seriola dumerili).Using dual-luciferase reporter assays,we demonstrate that wbp2 is a direct target of miR-133b,with miR-133b-3p binding to the 3'untranslated region(3'UTR)of wbp2.In vitro,miR-133b mimic significantly downregulate wbp2 expression,while the miR-133b inhibitor increase wbp2 levels.Consistently,in vivo,wbp2 expression is upregulated following antagomir-133b treatment and downregulated following agomir-133b.RNA fluorescence in situ hybridization(RNA-FISH)results reveal miR-133b and wbp2 co-localization in ovarian interstitial cells.Notably,phylogenetic analysis indicates that miR-133b-3p and wbp2 are highly conserved among bony fish species.Additionally,dual-luciferase assays in other bony fish species including Oreochromis niloticus and Danio rerio also confirm the targeting effect of miR-133b-3p on wbp2,suggesting that this regulatory mechanism is conserved across bony fish.This research provides a theoretical foundation for further exploration of non-coding RNA-mediated regulation in gonadal development in teleost.展开更多
基金The Fund of Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang)under contract No.ZJW-2023-01the Guangdong Basic and Applied Basic Research Foundation under contract Nos 2023A1515010576 and 2024A1515012859the Marine Youth Talent Project of Zhanjiang under contract No.2023E0006.
文摘Sex-biased microRNAs(miRNAs)influence gonadal development in fish by directly targeting genes associated with estrogen production pathways.WW domain-binding protein 2(WBP2)functions as a crucial transcriptional co-activator of the estrogen and progesterone receptors(PGR).This study investigates the direct modulation of a sex-biased miR-133b on wbp2 and its regulatory role in gonadal development in fish,the greater amberjack(Seriola dumerili).Using dual-luciferase reporter assays,we demonstrate that wbp2 is a direct target of miR-133b,with miR-133b-3p binding to the 3'untranslated region(3'UTR)of wbp2.In vitro,miR-133b mimic significantly downregulate wbp2 expression,while the miR-133b inhibitor increase wbp2 levels.Consistently,in vivo,wbp2 expression is upregulated following antagomir-133b treatment and downregulated following agomir-133b.RNA fluorescence in situ hybridization(RNA-FISH)results reveal miR-133b and wbp2 co-localization in ovarian interstitial cells.Notably,phylogenetic analysis indicates that miR-133b-3p and wbp2 are highly conserved among bony fish species.Additionally,dual-luciferase assays in other bony fish species including Oreochromis niloticus and Danio rerio also confirm the targeting effect of miR-133b-3p on wbp2,suggesting that this regulatory mechanism is conserved across bony fish.This research provides a theoretical foundation for further exploration of non-coding RNA-mediated regulation in gonadal development in teleost.
