A Cathepsin B Inhibitor, E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos
نویسندگان
چکیده
Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.
منابع مشابه
I-12: Nuclear Reprogramming in Bovin Somatic Cell Nuclear Transfer
Somatic cell nuclear transfer (SCNT or cloning) returns a differentiated cell to a totipotent status; a process termed nuclear reprogramming. Reproductive cloning has potential applications in both agriculture and biomedicine, but is limited by low efficiency. To understand the deficiencies of nuclear reprogramming, our research has focused on both candidate genes and global gene expression pat...
متن کاملP-115: Melatonin Increases Developmental Rate of In Vitro Mouse Somatic Cell Nuclear
Background: The beneficial effect of supplementing culture medium with melatonin has been reported during in vitro embryo development of species such as mouse, bovine and porcine. However, the effect of melatonin on the mouse somatic cell nuclear transfer remained unknown. Materials and Methods: In this study, we assessed the effects of various concentrations of melatonin (10-6 to 10-12 M) on t...
متن کاملO-7: Improved In Vitro Development of Cloned Bovine Embryos Using S-Adenosylhomocysteine,A Non-Toxic Epigenetic
Background: Development of cloned bovine embryos. Materials and Methods: Oocytes collection,oocyte denudation, oocyte enucleation, nuclear transfer, electrofusion, reconstructed embryo activation, culture of reconstructed and IVF embryo,and treatment with SAH post fusion interval Treatment of reconstructed embryos with TSA for 12 hours after activation, preparation of somatic donor cells, donor...
متن کاملIn vitro Development of Interspecies Somatic Cell Nuclear Transfer Embryos Derived from Murine Embryonic Fibroblasts and Bovine Oocytes
Interspecies somatic cell nuclear transfer (iSCNT) is a useful method to preserve endangered species and to study the reprogramming event of a nuclear donor cell by the oocyte. Although several studies of iSCNT using murine cells and bovine oocytes have been reported, the development of murine-bovine iSCNT embryos beyond the 8-cell stage has not been successful. In this paper, we examined the d...
متن کاملInhibition of cathepsin B activity reduces apoptosis by preventing cytochrome c release from mitochondria in porcine parthenotes
Cathepsin B, a lysosomal cysteine protease of the papain family, has recently been implicated in the quality and developmental competence of bovine preimplantation embryos. In this study, to determine whether inhibition of cathepsin B activity can improve porcine oocyte maturation and early embryo developmental competence, we supplemented in vitro maturation or embryo culture media with E-64, a...
متن کامل