Effect On Activity Of Ovarian Granulosa Cells By MiR-10b And BDNF In Dairy Goats

MicroRNAs (miRNAs), as small noncoding RNAs, were recently found to regulate geneexpression at post-transcriptional levels. It has been noticed that miRNAs play an importantrole on the development of ovary, fallopian tube, uterus and other tissues or organs related toreproductive performance. Studying the function of miRNAs and target genes in estrousovarian tissue is one of hot spots in current animal reproduction research.This experiment hadselected differential expressed miR-10b from the constructed miRNA banks of estrous ovariantissue from polytocous(3-5) and monotocous dairy goats, using bioinformatics and real timePCR technologies to study the function of miR-10b in ovarian granular cells(GCs) activity, andidentify the target genes. The main results are as follows:1. Differential expressed miR-10b had been selected from the constructed miRNA banks ofestrous ovarian tissue from polytocous(3-5) and monotocous dairy goats by bioinformaticanalysis. We also used real-time PCR technology to detect the relative expression of miR-10bfrom the miRNA banks, the result showed that the relative expression of miR-10b wassignificantly upregulated in uniparous dairy goats compared with multiparous dairy goat.2. Prediction and identification of miR-10b target genesBDNF had been predicted as the target gene of miR-10b by bioinformatic analysis. Then,we idendified that BDNF is one of the target genes of miR-10b by luciferase analysis andreal-time PCR technology.3. In the current study, we cloned the full coding region of BDNF gene with753nucleotides and250amino acids. The amino acid sequence of the caprine BDNF proteinshared100%and99%homology compared to predicted caprine and bovine, respectively.4. Adding BDNF factor in cell medium promoted the activity of ovarian GCs. High-levelmiR-10b suppressed the activity of ovarian GCs. Adding BDNF factor and overexpressedmiR-10b at the same time, activity of ovarian GCs had no significant changes.Results showed that miR-10b can suppress GCs activity by targeting BDNF whichprovided experimental basis for study the caprine follicular development.