miR ‐34 regulates reproduction by inhibiting the expression of MIH, CHH, EcR, and FAMeT genes in mud crab Scylla paramamosain

In this study, we used bioinformatic tools to identify the target genes of miR‐34 in eyestalk. Six reproduction‐related genes wi th an intact 3′‐untranslated region (UTR), including molt‐inhibiting hormone (MIH), crustacean hyperglycemic hormone (CHH), vitellogenesis‐inhibiting hormone, red pigment concentrating hormone, ecdysone receptor (EcR), and farnesoic acid methyltransferase (FAMeT) were identified. When the 3′ ‐UTR plasmid vectors of the six genes were cotransfected with miR‐34 mimics into 293FT cells, respectively, the luciferase activities of four genes (MIH, CHH, EcR, and FAMeT) were significantly decreased compared with that in the control group; on the contrary, when the six plasmid vectors were cotransfected with the miR‐34 inhibitor respectively, the luciferase activities of four genes (MIH, CHH, EcR, and FAMeT) were significantly higher than that in the control group. When agomiR‐34 and antagomiR‐34 were injected into the eyestalk respectively in vivo, the expression levels of the MIH, CHH, EcR, and FAMeT genes were detected by a quantitative real‐time polymerase chain reaction. The results showed that agomiR‐34 suppressed the expression of the four genes, whereas antagomiR‐34 enhanced their expression. These experimental results confirmed our hypothesis that miR‐34 m ay indirectly regulate reproduction via binding to the 3′‐UTRs of MIH, CHH, EcR, and FAMeT genes and suppressing their expression.
Source: Molecular Reproduction and Development - Category: Reproduction Medicine Authors: Tags: RESEARCH ARTICLE Source Type: research