Abstract
Abstract
UV-irradiation of oocytes during enucleation and serum starvation of donor cells during cell cycle synchronization may compromise the development competence of cloned embryos through excessive generation of reactive oxygen species (ROS). Here, we show that 3-hyroxyflavone (a flavonoid having hydroxyl group at 3 carbon position) inhibits UV- and serum starvation-induced ROS production in oocytes and donor cells, respectively, and thereby improves the in vitro development of cloned porcine embryos (p<0.05). In a parthenogenetic model, UV-irradiation for 5 sec or more was found to reduce the in vitro development and quality of the embryo, which could be rescued by their culture in the presence of 3-hydroxyflavone. The rescuing effect of 3-hydroxyflavone was associated with significant reduction in ROS level (14.4±1.0 vs. 47.1±6.7), increase in ERK signaling molecules by 2.1-fold, and decrease in Caspase3 expression by 3.2-fold. Culture of donor cells (18.5±1.4 vs. 13.0±1.7%) or cloned embryos (20.6±1.1 vs. 12.2±1.1%) in the presence of 3-hydroxflavone also increased (p<0.05) the rates of blastocyst formation in cloned embryos produced by the nuclear transfer of serum-starved donor cells into recipient cytoplasts exposed to UV-irradiation during the enucleation step. Importantly, both parthenotes and cloned embryos cultured in the presence of 3-hydroxyflavone had significantly increased ability to expand, and contained a higher number of cells than those of the control group (p<0.05). These results suggest that 3-hydroxyflavone may be useful for improving the in vitro developmental potential of cloned embryos through inhibition of ROS production induced by the UV-irradiation of oocyte and/or the serum starvation of donor cells.
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