Presence of Cleaved Caspase 3 in Swine Embryos of Different Developmental Capacities Produced by Parthenogenetic Activation

This study assessed the presence of cleaved caspase 3 (CC3) during the in vitro development of swine embryos produced by parthenogenetic activation (PA). Embryos with high and low capacity to develop into blastocysts and the exposure to a caspase inhibitor (z-DEVD-fmk) were used to investigate the effect of CC3 on embryo development. The blastocyst rate (64.3% vs. 16.4%) and the average number of nuclei per blastocyst (39.7 vs. 19.8) were significantly higher (P< 0.05) in early( before 24 hr) compared to late-(between 24 and 48 hr) cleaving embryos after PA. CC3 was mainly detected in the cytoplasm of Day-2 and -4 embryos, but was primarily localized in the nucleus of Day-5 and -6 embryos. The fluorescence signal for CC3 relative to negative controls was significantly higher (P< 0.05) in early-(2.42-fold) compared to late-cleaving (1.39-fold) embryos at day 2 of culture. Treatment with z-DEVD-fmk during the first 24 or 48 hr of the culture period resulted in more embryos developing into blastocysts compared to the control group (55.8% and 55.1% vs. 37%, respectively; P< 0.05). This study confirmed the presence of CC3 in PA embryos from the two-cell to the blastocyst stage, and revealed that CC3 cellular-localization changed during embryo development. CC3 was shown to be more abundant in early-cleaving and more developmentally competent embryos compared to late-cleaving and less developmentally competent embryos. The inhibition of caspase activity at the beginning, but not at the end, of the culture period affected development of PA embryos. Mol. Reprod. Dev. 78: 673-683, 2011. (C) 2011 Wiley-Liss, Inc.