Comparisons Of Expression Level Of IFRG15 Among Pre-implantation Embryo, Male And Female Mouse Tissues At Different Growth Stages

Mouse IFRG15 is interferon-a responsive genes, coding a protein contain 131 amino acid to inhibit the reproduction of virus after modified. Research has shown that IFRG15 is differentiated expression in mouse 2-cell cloned embryos and rabbit pre-implantation cloned embryos, then we can inferred that IFRG15 differentiated expression may be one of the reasons cause the low Somatic cells cloned embryonic development rate, but no reports about the expression rules of mouse pre-implantation embryo and in different issues after birth. This experiment use Real-time PCR to study the expression rules of mouse IFRG15 expression in pre-implantation embryos in vivo,in vitro fertilized and in some tissues of every period after birth in mRNA level. Preliminary explored the relationship between its expression quantity and his-tone acetylation in pre-implantation embryo.First we comparison laboratory existing two embryos in vitro culture system. Then, establish best in vitro fertilization system by control some conditions (the cap-citation time of sperm, age of Oocytes, cumulus cells with or not). Treat pronuclear zygotes with TSA to obtain histone acetylation abnormal embryos. Analysis the expression rule of IFRG15 among the pre-implantation embryos in vivo,in vitro fertilized and TSA treated by real-time PCR. Analysis 7 tissues expression rule of IFRG15 during mouse growth cycle by step-one RT-PCR.After the study we can inform that under current conditions in the laboratory CZB medium is suitable for in vitro mouse embryos culture. Improved the key step of in vitro fertilization on the basis of predecessors, put 1.5h capacitated sperm and COCs after injected hCG 13.5h is best for quality of embryos. The result of real-time PCR demonstrates that expression quantity of IFRG15 is highest at 4-cell stage. At the same time we found the expression quantity in the group treat with TSA is higher than the normal group. So we can demonstrate that high histone acetylation is up regulation for expression of IFRG15. Tracking detection for 7 different tissues after mouse birth, we find the expression of IFRG15 exist selective expression differentiated in lung and reproductive organs.The study first time analysis mouse IFRG15 expression rule of pre-implantation embryos in vivo,in vitro and the embryos treat with TSA. It provides a reference for improve the condition of culture in vitro, for improving the development quantity of the embryos culture in vitro and revealing the role in mouse IFRG15 in pre-implantation embryos. At the same time, it has detected the mouse IFRG15 expression level of the 7 issues of different life periods of male and female. We got the gene expression patterns in the later process of development for the first time; also lay a foundation for the future study of this gene's function.