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Generation of Reactive Oxygen Species in Bovine Somatic Cell Nuclear Transfer Embryos during Micromanipulation Procedures  

Hwang, In-Sun (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Bae, Hyo-Kyung (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
Cheong, Hee-Tae (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.36, no.1, 2012 , pp. 49-53 More about this Journal
Abstract
The present study was conducted to examine the generation of reactive oxygen species (ROS) during micromanipulation procedures in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine enucleated oocytes were electrofused with donor cells, activated by a combination of Ca-ionophore and 6-dimethylaminopurine culture. Oocytes and embryos were stained in dichlorodihydrofluorescein diacetate or 3'-(p-hydroxyphenyl) fluorescein dye and the $H_2O_2$ or $^.OH$ radical levels were measured. $In$ $vitro$ fertilization (IVF) was performed for controls. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each oocyte and embryo. The $H_2O_2$ and $^.OH$ radical levels of reconstituted oocytes were increased during manipulation (37.2~49.7 and 51.0~55.2 pixels, respectively) as compared to those of mature oocytes ($p$<0.05). During early $In$ $vitro$ culture, the ROS levels of SCNT embryos were significantly higher than those of IVF embryos ($p$<0.05). These results suggest that the cellular stress during micromanipulation procedures can generate the ROS in bovine SCNT embryos.
Keywords
Somatic cell nuclear transfer; Micromanipulation procedures; ROS generation; Cattle;
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