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KMID : 1023620120360010049
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Reproductive and Developmental Biology
2012 Volume.36 No. 1 p.49 ~ p.53
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Generation of Reactive Oxygen Species in Bovine Somatic Cell Nuclear Transfer Embryos during Micromanipulation Procedures
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Hwang In-Sun
Bae Hyo-Kyung
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 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 H2O2 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 H2O2 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.
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KEYWORD
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Somatic cell nuclear transfer, Micromanipulation procedures, ROS generation, Cattle
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