The Study Expression Of Antioxidant Enzyme In MET Period Of Mouse Embryos In Vivo And In Vitro

Studies have shown that early embryo in mice mother type to zygote type transition easily happened 2-cell block phenomenon,and the various factors in the external environment can cause elevated ROS content in embryo,antioxidant enzymes is the main enzymes remove intracellular ROS.As the research progress and the MET period includes 1-cell(27h),2 cell(42 h),2-cell(51 h)and 4 cell(66 h)),but for 2-cell,which period is the most prone to block are still unclear,and the change of the relationship between the main antioxidant enzymes of cell block phenomenon and ROS is unclear.Therefore,this experiment determined the ROS content of each period through the DCFH-DA,the PCR and the Western blot,to observe the relationship between antioxidant enzymes and ROS.The result is as follows:1.From of DCFH-DA,the results showed that the level of ROS in the MET of embryo in the later 2-cell was highest,significantly higher than that of the stage of 1-cell,middle 2-cell and 4-cell.And the vivo group was significantly higher than that of in vitro group.2.The gene mRNA expression of different antioxidant enzymes in 1-cell to 4-cell period in the early embryonic development are different,the expression of GPX1 gene in later 2-cell in mouse embryo was the highest,significantly than that the stage of 1-cell,middle 2-cell and 4-cell.The expression of and SOD1 and SOD2 and SOD3 CAT and GPX2 gene in 4-cell stage in mouse embryo was the highest,significantly than that the stage of 1-cell and 2-cell.3.The expression level of CAT and GPX1 and SOD1 gene in the MET of embryo in vitro group were significantly higher than that of in vivo group,the mRNA expression level of SOD2 and SOD3 gene in the MET of embryo were increase trend,and the mRNA expression level of SOD2 in 1-cell and middle 2-cell were no significant difference in vitro group and in vivo group,the mRNA expression level of SOD3 in later 2-cell and 4-cell were no significant difference in vitro group and in vivo group,the expression level of GPX2 gene in the period of MET had no significant difference in vivo and in vitro.4.With protein immunoblot results show that the protein expression level of CAT and SOD1 gene in 4-cell significantly higher than other groups,the protein expression level of GPX1 gene in latter 2-cell significantly higher than other groups.The protein expression level of SOD1 and GPX1 in vitro group was significantly higher than that of in vivo group,but the protein expression level of CAT in later 2-cell was no significantly difference in vivo and in vitro,description the transcription and translation cannot be synchronized in the zygotic genome.Conclusion:During the MET period of the mice,later 2-cell is a major period of cell blocking.In mice,the CAT and the SOD1 gene are the main factors that remove the internal ROS and overcome cell blocking.