title: Presence of cleaved caspase 3 in swine embryos of different developmental capacities produced by parthenogenetic activation.
abstract: 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.
authors: Departamento de Reprodução Animal, Universidade de São Paulo, SP, Brazil.
authors: Ana Rita S
authors: AR
authors: Coutinho
authors: Mayra E O
authors: ME
authors: Assumpção
authors: Vilceu
authors: V
authors: Bordignon
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001755:Blastocyst; D053148:Caspase 3 / Q000378:metabolism*; D061945:Caspase Inhibitors; D002467:Cell Nucleus; D016009:Chi-Square Distribution; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D005260:Female; D006651:Histocytochemistry; D009842:Oligopeptides / Q000494:pharmacology; D010312:Parthenogenesis / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D013552:Swine / Q000196:embryology* / Q000378:metabolism*
chemical_list: D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; D053148:Caspase 3