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Effects of cycloheximide or 6-dimethyl aminopurine on the parthenogenetic activation of pig oocytes using pulsatile treatment with nitric oxide donor

Author

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  • T. Krejčová

    (Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Prague, Czech Republic)

  • J. Petr

    (Research Institute of Animal Production, Prague-Uhříněves, Czech Republic)

  • M. Krejčová

    (Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Prague, Czech Republic)

  • K. Kheilová

    (Department of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Prague, Czech Republic)

Abstract

Pig oocytes matured in vitro were parthenogenetically activated using nitric oxide donor SNAP (2mM). Continuous treatment successfully activated the oocytes only after more than 12 hours of exposure. Pulsatile treatments during which oocytes were repeatedly exposed to 2mM SNAP for a short time (10, 20 or 30 minutes) were more efficient with regard to the activation rate, even when the total exposure time did not exceed 4 hours. Parthenogenetic development was very limited after continuous treatment with 2mM SNAP. A significantly higher proportion of developing parthenogenetic embryos was observed after the pulsatile treatment (development to the morula stage 0 vs. 18%; development to the blastocyst 0 vs. 7%; P < 0.05). However, this developmental rate was significantly lower (P < 0.05) than the development induced by conventional activation treatment with calcium ionophore (development to the morula stage, 23%; development to the blastocyst stage, 18%). When we combined pulsatile SNAP-treatment with the effect of protein kinase inhibitor 6-dimethyl aminopurine (6-DMAP) (2mM 6-DMAP for 2 hours) or with the inhibitor of protein synthesis cycloheximide (CHX) (10 µM CHX for 2 hours), we observed a significant increase (P < 0.05) in the activation rate when compared to the respective pulsatile SNAP-treatment without 6-DMAP or CHX (63 vs. 78% of activated oocytes for 6-DMAP; 63 vs. 83% of activated oocytes for CHX). However, the development of parthenogenetic embryos was not enhanced when the pulsatile SNAP-treatment was combined with 6-DMAP or with CHX.

Suggested Citation

  • T. Krejčová & J. Petr & M. Krejčová & K. Kheilová, 2009. "Effects of cycloheximide or 6-dimethyl aminopurine on the parthenogenetic activation of pig oocytes using pulsatile treatment with nitric oxide donor," Czech Journal of Animal Science, Czech Academy of Agricultural Sciences, vol. 54(7), pages 293-306.
    • Handle: RePEc:caa:jnlcjs:v:54:y:2009:i:7:id:1724-cjas
    DOI: 10.17221/1724-CJAS
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    References listed on IDEAS

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    1. Richard C. Kuo & Gregory T. Baxter & Stuart H. Thompson & Stephen A. Stricker & Chris Patton & Joseph Bonaventura & David Epel, 2000. "NO is necessary and sufficient for egg activation at fertilization," Nature, Nature, vol. 406(6796), pages 633-636, August.
    2. J. Petr & E. Chmelíková & A. Dörflerová & M. Ješeta & Z. Kuthanová, 2007. "Effects of protein kinase C on parthenogenetic activation of pig oocytes using calcium ionophore or nitric oxide-donor," Czech Journal of Animal Science, Czech Academy of Agricultural Sciences, vol. 52(12), pages 415-422.
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    Keywords

    activation; nitric oxide; oocyte; pig;
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