Effect of Kinetin on In Vitro Development of Parthenogenetic Porcine Oocytes Exposed to Demecolcine Prior to Activation

  • Kim, Ki-Young (Laboratory of Embryo Biotechnology and Cellular Reprogramming, Dental Research Institute and BK2I CLS, Seoul National University School of Dentistry) ;
  • Park, Sang-Kyu (Laboratory of Embryo Biotechnology and Cellular Reprogramming, Dental Research Institute and BK2I CLS, Seoul National University School of Dentistry) ;
  • Roh, Sang-Ho (Laboratory of Embryo Biotechnology and Cellular Reprogramming, Dental Research Institute and BK2I CLS, Seoul National University School of Dentistry)
  • Published : 2009.06.30

Abstract

This study was designed to investigate the effect of kinetin on in vitro development of parthenogenetic porcine oocytes exposed to demecolcine prior to activation. In vitro matured metaphase II stage oocytes were incubated in 0 or 2 ${\mu}$g/ml demecolcine supplemented defined culture medium for 3 h and the oocytes were activated electrically. The parthenogenetic porcine embryos were then cultured in 0 or 200 ${\mu}$M kinetin supplemented defined culture medium for 7 days. Regardless of demecolcine treatment, kinetin supplementation increased blastocyst rates significantly (7.0% versus 12.1% and 4.9% versus 8.5%; Control versus Kinetin and Demecolcine versus Kinetin + Demecolcine, respectively, p<0.05). Demecolcine treatment before activation tended to decrease blastocyst rates regardless of kinetin supplementation although it is not statistically significant. Total cell numbers in the blastocysts also tended to be elevated in embryos when supplemented with kinetin, however only the result between Kinetin and Demecolcine groups is statistically significant (37.6 ${\times}$ 7.2 versus 28.1 ${\times}$ 9.5, respectively, p<0.05). In conclusion, the present report shows that kinetin enhances developmental competence of parthenogenetic porcine embryo regardless of demecolcine pre-treatment before parthenogenetic activation when they were developed in defined culture condition.

Keywords

References

  1. Barciszewski J, Siboska GE, Pedersen BO, Clark BF and Rattan SI. 1997. A mechanism for the in vivo formation of $N^6$-furfuryladenine, kinetin, as a secondary oxidative damage product of DNA. FEBS Lett. 414:457-460 https://doi.org/10.1016/S0014-5793(97)01037-5
  2. Bayliss M. 1985. Control of cell division in cultured cells. In:Bryant JA, Francis D, editors. The Cell Division Cycle in Plants., Cambridge University Press, Cambridge, pp. 157-178
  3. Francis D and Sorrell DA. 2001. The interface between the cell cycle and plant growth regulators: A mini review. Plant Growth Regul. 33:1-12 https://doi.org/10.1023/A:1010762111585
  4. Gasparrini B, Gao S, Ainslie A, Fletcher J, McGarry M, Ritchie WA, Springbett AJ, Overstrom EW, Wilmut I and De Sousa PA. 2003. Cloned mice derived from embryonic stem cell karyoplasts and activated cytoplasts prepared by induced enucleation. BioI. Reprod. 68:1259-1266 https://doi.org/10.1095/biolreprod.102.008730
  5. Miller CO, Skoog F, Okumura FS, von Saltza MH and Strong FM. 1956. Isolation, structure and synthesis of kinetin, a substance promoting cell division. J. Am. Chem. Soc. 78:1375-1380 https://doi.org/10.1021/ja01588a032
  6. Miller CO, Skoog F, von Saltza MH and Strong FM. 1955. Kinetin, a cell division factor from deoxyribonucleic acid. J. Am. Chem. Soc. 77:1329-1334
  7. Olsen A, Siboska GE, Clark BF and Rattan SI. 1999. N(6)-Furfuryladenine, kinetin, protects against Fenton reaction-mediated oxidative damage to DNA. Biochem. Biophys. Res. Commun. 265:499-502 https://doi.org/10.1006/bbrc.1999.1669
  8. Rattan SI and Clark BF. 1994. Kinetin delays the onset of ageing characteristics in human fibroblasts. Biochem. Biophys. Res. Commun. 201:665-672 https://doi.org/10.1006/bbrc.1994.1752
  9. Rieder CL and Palazzo RE. 1992. Colcemid and the mitotic cycle. J. Cell. Sci. 102(3):387-392
  10. Rodriguez-Arnaiz R, Sortibran AC and Tellez GO. 2004. Detection of mitotic recombination and sex chromosome loss induced by adriamycin, chlorambucil, demecolcine, paclitaxel and vinblastine in somatic cells of Drosophila melanogaster in vivo. Mutagenesis 19:121-127 https://doi.org/10.1093/mutage/geh013
  11. Sheu JR, Hsiao G, Shen MY, Chou CY, Lin CH, Chen TF and Chou DS. 2003. Inhibitory mechanisms of kinetin, a plant growth-promoting hormone, in platelet aggregation. Platelets. 14:189-196 https://doi.org/10.1080/0953710021000060925
  12. Tani T, Shimada H, Kato Y and Tsunoda Y. 2006. Demecolcine-assisted enucleation for bovine cloning. Cloning Stem Cells 8:61-66 https://doi.org/10.1089/clo.2006.8.61
  13. Won C, Park SK, Cho SG, Min BM and Roh S. 2008. Kinetin enhances in vitro development of parthenogenetic and nuclear transfer porcine embryos. Mol. Reprod. Dev. 75:1701-1709 https://doi.org/10.1002/mrd.20920
  14. Yin XJ, Kato Y and Tsunoda Y. 2002a. Effect of enucleation procedures and maturation conditions on the development of nuclear-transferred rabbit oocytes receiving male fibroblast cells. Reproduction 124:41-47 https://doi.org/10.1530/rep.0.1240041
  15. Yin XJ, Tani T, Yonemura I, Kawakami M, Miyamoto K, Hasegawa R, Kato Y and Tsunoda Y. 2002b. Production of cloned pigs from adult somatic cells by chemically assisted removal of maternal chromosomes. BioI. Reprod. 67:442-446 https://doi.org/10.1095/biolreprod67.2.442