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KMID : 0900920030270020103
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Korean journal of Animal Reproduction
2003 Volume.27 No. 2 p.103 ~ p.113
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Study of Efficient Production of Cloned Embryos in Porcine 1. Effect of Ethanol, Ca^{2+} Ionophore, 6-DMAP, and Cycloheximide Concentration and Exposure Time on Activation, Cleavage, and In Vitro Development of Porcine Oocytes
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Kim Kwang-Hyun
Kang Man-Jong
Moon Seung-Ju
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Abstract
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This study was conducted to investigate the optimal condition for produce of large quantity recipient oocytes on porcine cloned embryos. In order to determined the optimum concentration and exposure time of ethanol, Ca^{2+}-ionophore, 6-DMAP and cycloheximide, in vitro matured oocytes were activated in TCM-199 containing various chemicals and 15% FBS. The activated oocytes were cultured in Whitten’s medium containing 10% FBS at 5% CO_2. 1. When the porcine oocytes were activated with the ethanol, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 10% for 10 minutes, which was significantly higher than all of the other treatment(53.4%, 51.6% and 39.9%, respectively). 2. When the porcine oocytes were activated with the Ca^{2+}-ionophore, the pronucleus furmation, cleavage, and in vitro development rate were found significantly higher in the 25muM fur 2min. treatment than those of other concentration and exposure time(59.7%, 62.2% and 43.9%, respectively). 3. When the porcine oocytes were activated with the 6-DMAP, the best pronucleus formation, cleavage, and in vitro development rate were obtained in the 2mM for 2hr~4.5hr(57.3%, 58.4% and 29.0%, respectively). 4. When the porcine oocytes were activated with the cycloheximide, result showed that pronucleus formation, cleavage, and in vifro development rate were 52.1%, 47.7% and 31.8%, respectively, in the 5mutextrm{g}/ml for 4hr~6hr treatmrent, which was significantly higher than all of the other treatment. These results suggested that the active condition of porcine oocytes was established by optimum concentration and exposure time among different chemicals for produce of large quantity recipient oocytes.s.
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KEYWORD
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Ethanol, Ca^{2+}-ionophore, 6-DMAP, Cycloheximide, Activation
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