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.

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