Regulation of vtg and VtgR in mud crab Scylla paramamosain by miR-34.

Abstract

Vitellogenin (Vtg) is the precursor of major yolk protein and plays a crucial role in the maturation of oocytes and the production of eggs in oviparous animals. Vitellogenin receptor (VtgR) mediates the transport of Vtg explicitly to oocytes in the membrane. In a previous study, we found that miR-34 can regulate the expression of some eyestalk genes and affect reproduction in mud crab Scylla paramamosain, one of the most important economic crabs on the coasts of southern China. In this study, firstly, we found that miR-34 can target at 3'-UTR of Vtg and VtgR genes by using bioinformatic tools and predicted miR-34 might depress the expression of Vtg and VtgR. Secondly, the relative luciferase activity of HEK293T cells co-transfected with miRNA mimic and pmir-RB-REPORTTM-Vtg/VtgR-3'UTR was significantly lower than those of cells co-transfected with mimic NC and pmir-RB-REPORTTM-Vtg/VtgR-3'UTR. Finally, in vivo experiments showed that agomiR-34 could repress the expression of Vtg and VtgR genes, while Antigomir-34 could promote the expression of these two genes. These results confirm our hypothesis and previous published results that miR-34 may indirectly regulate ovarian development by binding to the 3'-UTR of Vtg and VtgR genes and inhibiting their expression.