Effect Of Different Does Of Hormones On Development Of Mouse Implantation Embryo And Frozen-Thawed Embryo

Mouse is the one of the important model animal for the research or application fields in embryo research, genetic research and human disease. To expand research fields and provide a reference for animal breeding, a high-efficiency technology platform of mouse embryo has recently become a main objective for our lab. With different dosage of hormone (5IU PMSG+5IU HCG and10IU PMSG+10IU HCG) for superovulation of Kunming mice by intraperitoneal injection, this study armed to investigate the effects of the different treatment on subsequent development of mouse implantation embryo and frozen-thawed embryos and changes of proteins in ovary and serum. Accumulation of these data can lay the foundation to establish an efficient operating platform for mouse embryo.Firstly, the optimum culture medium was determined. After the mice embryo have been cultured in KSOM, M16and CZB medium for4days, The blastocyst rate in KSOM medium was significant higher than in M16and CZB medium (75.94%vs46.07%,63.93%, P<0.05). So KSOM medium was used in subsequent experiments. Secondly, the effect of different concentration of taurine in KSOM medium for the development of mouse preimplantation embryos (OmM,2.5mM,5mM,10mM and50mM) has been investigated. The results showed that the blastocyst rates showed no significant differences among the five groups (75.76%、77.45%、73、56%、73.06%、71.78%, respectively.)(P>0.05).Finally, KunMing mice were treated at different dose of hormone to superrovulation (5IU PMSG+5IU HCG and10IU PMSG+10IU HCG), and the blastocyst rates were significant difference between the two groups (76.45%and67.08%P<0.05).After KunMing mice were treated at different dose of hormone to superrovulation (5IU PMSG+5IU HCG and10IU PMSG+10IU HCG), Tissue specific expression protein in serum and ovarian sample grinded in liquid nitrogen has been detected with flight mass spectrometry (MALDI-TOF/TOF). In serum samples, specific expression protein gi19116236was detected in control group, and specific expression protein gi19116236and gil48669394were detected in treatment group with5IU PMSG+5IU HCG, but no specific expression protein was detected in treatment group with10IU PMSG+10IU HCG. There was no specific protein in the ovarian sample detection.After KunMing mice were treated at different dose of hormone to superrovulation (5IU PMSG+5IU HCG and10IU PMSG+10IU HCG), the embryos from the two treatment groups were frozen and then thawed. The result showed that embryonic recovery rate and blastocyst rate in the treatment group with10IU PMSG+IOIU HCG were significantly higher than in the treatment group with5IU PMSG+5IU HCG.(86.03%,73.63%VS82.93%,67.95%,P<0.05)Inconclusion, the data from this research showed that the KSOM medium was suitable for kunming mice preimplantation embryo culture.5IU PMSG+5IU HCG combined hormones were suitable for mice preimplantation embryo culture.10IU PMSG+10IU HCG combination were more suitable for the embryo cryopreservation than5IU PMSG+5IU HCG combination. Specific expression proteins showed difference in different treatments with different hormones combination,5IU PMSG+5IU HCG combination hormones promote the release of gi148669394protein in serum of mice,10IU PMSG+10IU HCG combination hormones inhibit the release of gi148669394and gi19116236proteins in serum of mice, but the structure and function of gi19116236and gi148669394remained unclear.