Inactivated Sendai-virus-mediated fusion improves early development of cloned bovine embryos by avoiding endoplasmic-reticulum-stress-associated apoptosis
- 1 January 2011
- journal article
- research article
- Published by CSIRO Publishing in Reproduction, Fertility and Development
- Vol. 23 (6), 826-836
- https://doi.org/10.1071/rd10194
Abstract
Somatic cell nuclear transfer (SCNT) is a powerful tool, not only for producing cloned animals, but also in revealing various early developmental events. However, relatively little is known regarding the biological events and underlying mechanism(s) directly associated with early development of SCNT embryos. Here, we show that production of high-quality bovine SCNT blastocysts is dependent on the method used for fusion and the associated reduction in endoplasmic reticulum (ER) stress. During fusion between the donor cell and the enucleated oocyte, electrofusion triggers spontaneous oocyte activation, accompanied by an increase in intracellular Ca2+ and improper nuclear remodelling. These events can be greatly reduced by the use of Sendai virus (SV)-mediated fusion. Moreover, SV-SCNT improves the blastulation rate and blastocyst quality, defined by the number and ratio of inner cell mass and trophectoderm cells in each blastocyst, in comparison with electrofusion-mediated SCNT (E-SCNT). Interestingly, expression of ER-stress-associated genes and blastomere apoptosis were significantly increased in E-SCNT embryos. These increases could be reversed by inhibition of ER stress or by using the SV-mediated fusion method. Collectively, these results indicate that SV-mediated fusion improves the developmental competence and quality of SCNT blastocysts, by reducing ER-stress-associated apoptosis.Keywords
This publication has 39 references indexed in Scilit:
- Nature, 2009
- Nucleologenesis and embryonic genome activation are defective in interspecies cloned embryos between bovine ooplasm and rhesus monkey somatic cellsBMC Developmental Biology, 2009
- The Unfolded Protein Response Contributes to Preimplantation Mouse Embryo Death in the DDK Syndrome1Biology of Reproduction, 2009
- Role of endoplasmic reticulum stress in acrolein-induced endothelial activationToxicology and Applied Pharmacology, 2009
- Inheritable Histone H4 Acetylation of Somatic Chromatins in Cloned EmbryosJournal of Biological Chemistry, 2006
- Endoplasmic reticulum stress: cell life and death decisionsJCI Insight, 2005
- Fertilization and InsP3-induced Ca2+ release stimulate a persistent increase in the rate of degradation of cyclin B1 specifically in mature mouse oocytesDevelopmental Biology, 2004
- Comparison of Microinjection (Piezo-Electric) and Cell Fusion for Nuclear Transfer Success with Different Cell Types in CattleCloning and Stem Cells, 2002
- Nuclear and microtubule reorganization in nuclear‐transferred bovine embryosMolecular Reproduction and Development, 2002
- Electrofusion‐induced intracellular Ca2+ flux and its effect on murine oocyte activationMolecular Reproduction and Development, 1992