Research Of Apoptosis Of Mouse Tetraploid Embryos And The Effect Of GSH Andβ-Mercaptoethanol To Apoptosis

staining. All of these are significant for the research of the early development of mouse tetraploid embryos and the acquirement of more quality tetraploid embryos. The results show:1. In this study, I prepared tetraploid embryos by electroporation fusion, blastocyst rate of diploid embryos, tetraploid embryos, add GSH or β-mercaptoethanol cultured the4N embryonic blastocysts were92.3%,80.6%,88.7%,85.3%. The rate of tetraploid embryo to blastocyst was significantly increased after adding glutathione or P-mercaptoethanol.2. TUNEL staining showed that apoptosis of tetraploid embryos mainly occured in the blastocyst stage, and the apoptosis rate was13.2%, significantly higher than the normal diploid embryos(4.1%). The apoptosis rate of tetraploid blastocyst was significantly reduced after adding glutathione or β-mercaptoethanol(8.5%and9.4%).3. PCNA immunofluorescence staining showed that the proliferative capacity of tetraploid embryos was lowest in the blastocyst stage, the PCNA positive rate of tetraploid blastocysts was73.8%, significantly lower than the diploid embryos (94.2%). After adding glutathione or P-mercaptoethanol to M16, PCNA positive rate of tetraploid blastocysts were85.5%and83.7%,were significantly improved..4. RT-PCR showed that, in the tetraploid embryo development process, the mRNA expression of the pro-apoptotic gene Bax in the blastocyst stage was significantly higher than that of the diploid embryos. The expresstion of Bax in the blastocyst stage was reduced after adding glutathione or β-mercaptoethanol. The expression of the antiapoptotic gene Bcl-2in tetraploid embryos had no significant changes in different developmental stages. The expression of Bcl-2in the morula and blastocyst stage was significantly increased after adding glutathione or β-mercaptoethanol. There is no significant changes in PCNA gene expression levels in diploid embryos during the tetraploid embryos.In tetraploid embryos, PCNA gene expression levels in two-cell stage to the morula stage shows no significant changes,in the blastocyst stage, the PCNA expression was reduced., PCNA expression was significantly increased in the blastocyst stage in tetraploid embryos that cultured in medium supplemented with glutathione or β-mercaptoethanol.Through the results above, we can see that tetraploid embryos apoptosis is ralated to bax, bcl-2gene, and the expression of PCNA reduced impact is also one of the reasons for its apoptosis. By improving culture conditions in medium supplemented with GSH or β-mercaptoethanol, can significantly improve the tetraploid embryos, reducing the occurrence of apoptosis.