The Preliminary Function Research Of MiR-370during Mouse Developmental Stage

MicroRNA is one of the important members of the22nt short single-strandnon-coding RNA family, which is a class of highly conserved in evolution endogenoussmall RNA molecules, widely exists in plants and animals. miR-370researched in thesubject locates on Dlk1-Dio3imprinted gene cluster, Gtl2downstream on mouse distalregion of chromosome12, whose mature body contains20nucleotides. As miR-370specifically expresses during embryonic development, researching deeply on itsabnormal expression is of great significance for mouse embryonic development,individual growth, disease-control activities.To research miR-370function in mouse development, in this study we build geneknock-in mouse model, using real-time quantitative PCR, HE staining and bisulfitesequencing methods to detect the miR-370expression, the morphology changes, DNAmethylation levels, observe sperm motility and estrous cycle. We hoped to analyze theimpact of various mouse tissues, especially the gonads and the impact on growth andsexual maturation of mouse embryonic development after overexpression of miR-370.Morphological observation showed that miR-370transgenic mice had less embryos,more unusual deaths and unbalanced sex ratio, and miR-370transgenic male micegained more weight than wide type mice. RT-PCR showed that miR-370expressionlevels have risen in transgenic mice tissues than in wild-type mice’s. Transgenic mouselung in the embryonic stage appear large number of congestion. Renal corticaldevelopment is well, cortex and medulla junction is not clear. The three boundaries ofplacental are not clear, sinus expansion and the presence of congestion. Transgenic micelung interstitial has severe bleeding with fusion alveolar fracture and local consolidation.Kidney blood vessels dilate with glomerular bleeding. Transgenic mouse testis emergethe phenomenon of seminiferous tubular atrophy and sperm cells decrease with reducedsperm motility, increased abnormal rate and the plasma membrane integrity ratedeclining. Transgenic mouse ovary granulosa cells loosely arranged with reducinglayers, accompanied by estrous cycle disorders.Based on the above results that overexpression of miR-370has a huge impact onmouse embryo, placenta and adult tissues development. These results lay the foundationfor further elaboration functional miR-370in mouse development.