MiR-130a-3p And Target Gene PMEPA1 Regulate Steroidogenesis In Goat Ovarian Granulosa Cells

This study was based on the differentially expressed miRNA in the ovary libraries of the goat during the luteal phase and follicular phase.The ovarian granulosa cells?GCs? were treated with follicle-stimulating hormone to screen out the differentially expressed miRNA.It was planned to verify the target relationship between miRNA and the target genes at the cellular level to explore the effects of miRNA and target gene on the secretion of GCs steroid hormones.This provided basic data for exploring the genetic regulation of goat reproductive performance.In this experiment,goat ovaries were collected and ovarian GCs were isolated.The effects of miR-130a-3p and its target gene PMEPA1 on the secretion of estradiol?E₂? and progesterone from goat ovarian GCs were studied using fluorescence in situ hybridization,q PCR,WB,construction of lentiviral vectors,ELISA and dual-luciferase reporter assays.?1?The expression level of miR-130a-3p was significantly increased after 5IU of FSH was added to the primary goat GCs cultured in vitro,using the q PCR.miR-130a-3p had an obvious expression on GCs in the goat ovary,by the FISH.?2?The expression level of miR-130a-3p in GCs was increased significantly after the transfection of miR-130a-3p mimics in cells.On the contrary,that decreased significantly,after the infection inhibitor.E₂ and progesterone in the GCs supernatant were increased significantly after the transfection of miR-130a-3p mimics in the GCs,by ELISA.After the transfection of inhibitors in GCs,the results were reversed.?3?PMEPA1 was the target gene of miR-130a-3p,through the miRNA target gene prediction website,and the dual-luciferase reporter gene experiment.The expression level of PMEPA1 in GCs was suppressed when the cells were treated with miR-130a-3p mimics.?4?Goat's PMEPA1 interference lentivirus was successfully constructed.The expression of PMEPA1 in GCs was reduced.And the levels of E₂ and progesterone in the cell supernatant were also reduced when GC was infected with lentivirus.In summary,this study initially showed that miR-130a-3p has a targeted regulatory relationship with the goat PMEPA1.miR-130a-3p may regulate the secretion of estradiol and progesterone in goat granulosa cells by targeting PMEPA1.In addition miR-130a-3p and its target gene PMEPA1 may be involved in the development of goat follicles.