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KMID : 1023620130370010051
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Reproductive and Developmental Biology
2013 Volume.37 No. 1 p.51 ~ p.55
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Effect of Antioxidant Treatment during Parthenogenetic Activation Procedure on the Reactive Oxygen Species Levels and Development of the Porcine Parthenogenetic Embryos
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Bae Hyo-Kyung
Kim Soo-Hyun
Lee Sung-Young
Hwang In-Sun
Park Choon-Keun
Yang Boo-Keun
Cheong Hee-Tae
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Abstract
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The present study was conducted to examine the effect of antioxidant treatment during parthenogenetic activation procedure on the reactive oxygen species (ROS) levels and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by a combination of electric stimulus and 2 mM 6-dimethylaminopurine (6-DAMP) before in vitro culture. During the activation period, oocytes were treated with 50μM β-mercaptoethanol (β-ME), 100μM L-ascorbic acid (Vit. C) or 100μM L-glutathione (GSH). To examine the ROS level, porcine parthenogenetic embryos were stained in 10μM dichlorohydrofluorescein diacetate (H₂DCFDA) dye 20 h after culture, examined under a fluorescence microscope, and the fluorescence intensity (pixels) were analyzed in each embryo. The parthenogenetic embryos were cultured for 6 days to evaluate the in vitro development. The apoptosis was measured by TUNEL assay. The H₂O₂ levels of parthenogenetic embryos were significantly lower in antioxidant treatment groups (26.9±1.6~29.1±1.3 pixels/embryo, p<0.05) compared to control (33.2±1.7 pixels/embryo). The development rate to the blastocyst stage was increased in antioxidant treatment groups (32.0~32.5%) compared to control (26.9%, p<0.05), although, there was no difference in apoptosis among groups. The result suggests that antioxidant treatment during parthenogenetic activation procedure can inhibit the ROS generation and enhance the in vitro development of porcine parthenogenetic embryos.
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KEYWORD
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Antioxidant, ROS level, In vitro development, Parthenogenetic activation, Pig
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