Differential Expression And Function Of MiR-126in Bovine Corpus Luteum Development

Corpus luteum (CL) is well known for its role in animal breeding, including directeffects during estrous cycle alternation, embryo implantation, and pregnancymaintenance. CL mainly composed by the granulosa cells and thecal cells,it is atemporary secretory tissue that can be found in female mammals after ovulation.Therefore, to study the formation and development of the corpus luteum, improveanimal production value.A class of small, non-coding ribonucleic acids, termed microRNA (miRNA) hasrecently emerged as a new key player in the cellular control of gene expression. Byeither blocking translation or inducing target mRNA degradation.In recent years manyarticles reported, miR-126through a variety of ways to affect the integrity of theblood vessels and vascular Including many signaling pathways which closely relatedto angiogenesis, now really miR-126can be adjusted angiogenesis, the formation，development and degradation of the corpus luteum is a constantly accompanied by theprocess of angiogenesis and apoptosis.Therefore, miR-126and the luteal function,there may be a series of relations.In order to understand the link between miR-126and CL of the formation anddevelopment, using quantitative Real-Time RT-PCR method, detected in the lutealdifferent developmental periods of miR-126the relative expression level. Situhybridization experiments with digoxigenin-labeled miR-126probe in frozen sectionsof different developmental stages of the luteal experiments to validate the thereal-time RT-PCR results, and determine the law of the miR-126expression location.miRNA target genes predicted biological software (TargetScan, mfold, RNA22)applicated to determine the candidate target genes of miR-126. Luciferase reporterexperiments in NIH3T3cells, to ultimately determine the target genes of miR-126. Real-time RT-PCR experimental results show that, miR-126were expressed atdifferent developmental stages of CL, and in the latter part of the expression level ofthe highest early was the lowest, and there was a significant difference (p <0.05). Insitu hybridization experiments also proved that the miR-126expressed at differentdevelopmental stages of the CL, verify that the Real-time RT-PCR results.Apart fromthis it also showed that high miR-126expression in which the cytoplasm of lutealcells especially large luteal cell in the cytoplasm, on the contrary, within the nucleusalmost no expression.Dual luciferase gene report detection and fluorescence quantitative RT-PCRstudies have shown that EMP1and TLN2expressed in CL, and two target genes ofmiR-126target sites are not in the3’-UTR. miR-126in CL can regulate the expressionof these two genes.In summary, the results of this study show the law of miR-126expression andexpression position in different developmental stages of CL and, it’s also confirmed invitro, EMP1and TLN2is the two target genes of miR-126. The results of this studywill provide a theoretical basis for in-depth understanding of the molecular regulationof miR-126in the development and degeneration of the CL. The specific functionmiR-126in Reproduction, regulating mechanism as well as its animal fecundity, therole of animal fecundity still need further research to improve.