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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서지정보
ㆍ발행기관 : 한국동물번식학회
ㆍ수록지정보 : Reproductive & developmental biology / 37권 / 1호
ㆍ저자명 : Hyo-Kyung Bae, Soo-Hyun Kim, Sung-Young Lee, In-Sun Hwang, Choon-Keun Park, Boo-Keun Yang, Hee-Tae Cheong
목차
ABSTRACT INTRODUCTION MATERIALS AND METHODS In Vitro Maturation of Oocytes Parthenogenesis and In Vitro Culture Antioxidant Treatment Measurement of Intercellular ROS Content TUNEL Assay Statistical Analysis RESULTS H2O2 Levels of Parthenogenetic Embryos In Vitro Development of Parthenogenetic Embryos Apoptosis in Blastocysts DISCUSSION REFERENCES
영어 초록
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 50 μM β-mercaptoethanol (β-ME), 100 μM L-ascorbic acid (Vit. C) or 100 μM L-glutathione (GSH). To examine the ROS level, porcine parthenogenetic embryos were stained in 10 μM 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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