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Effects of Activation Methods on DNA Synthesis and Development of Parthenogenetic Porcine Embryos

Authors

  • SA Ock,

    1. College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea
    2. Institute of Animal Medicine, Gyeongsang National University, Jinju, Korea
    3. Department of Physiology, College of Medicine, Gyeongsang National University, Jinju, Korea
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  • DO Kwack,

    1. Division of Science Education, College of Education, Gyeongsang National University, Jinju, Korea
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  • B Mohana Kumar,

    1. College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea
    2. Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea
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  • J Han,

    1. Department of Physiology, College of Medicine, Gyeongsang National University, Jinju, Korea
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  • SW Kim,

    1. Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon, Korea
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  • GJ Rho

    1. College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea
    2. Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea
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Author’s address (for correspondence): G-J Rho, Department of Obstetrics and Theriogenology, College of Veterinary Medicine, Gyeongsang National University, 900 Gazwa, Jinju 660-701, Korea. E-mail: jinrho@gnu.ac.kr

Contents

This study investigated the timing of DNA synthesis and patterns of pronuclear (PN) formation during the first cell cycle, and its influence on developmental competence, velocity and proliferation index of porcine parthenote blastocysts produced by different activation treatments. Oocytes were activated as follows: electrical stimulation (EST), EST combined with 7.5 μg/ml cytochalasin B (EST + CCB), 10 μg/ml cycloheximide (EST + CHX) and 1.9 mm 6-dimethylaminopurine (EST + 6-DMAP) for 3 h. DNA synthesis and PN formation were evaluated using 1 mm 5′bromo-2′deoxy-uridne (BrdU) at 2 h intervals from 1 to 13 h or 5 to 13 h of post-activation (hpa), respectively. In EST, DNA synthesis started at 3 hpa, reached the peak at 11 hpa and decreased at 13 hpa. Treatment with 6-DMAP resulted in an early increase of DNA synthesis at 3 hpa, whereas CCB delayed DNA synthesis for 2 h. In EST and EST + 6-DMAP, most of the eggs showed 1PN, whereas, incidence of 2PN in EST + CCB was higher than 1PN. EST + CHX was observed with 1PN, 2PN and multiple PN. Blastocyst rate in EST + CCB and EST + 6-DMAP were significantly (p < 0.05) higher than EST + CHX. But, the developmental velocity was not different among groups. Proliferation index of blastocysts, as indicated the number of blastomere at S-phase of the cell cycle was low in all groups. In conclusion, CCB, CHX and 6-DMAP used for producing porcine parthenogenetic embryos induced different onset of DNA synthesis and PN, but they did not affect the subsequent embryo development.

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