Effect of Cysteamine on In Vitro Maturation, Fertilization and Culture of Porcine Oocytes

  • Yi, Y. J (Division of Animal Science & Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Kim, M. Y. (Division of Animal Science & Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Lee, S. H. (College of Visual Image & Health, Kongju National University) ;
  • T. S. Min (Department of Life Science, Korea Science and Engineering Foundation) ;
  • D. I. Jin (Division of Animal Science & Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University) ;
  • Park, C. S. (Division of Animal Science & Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University)
  • Published : 2003.12.01

Abstract

This study was carried out to investigate the effect of cysteamine addition during in vitro maturation, fertilization and culture of porcine oocytes. Oocytes were matured for the first 22 h in mTCM -199 media supplemented with or without 150 $\mu$M cysteamine. They then were matured for an additional 22 h in mTCM-199 media without hormones supplemented with or without 150 $\mu$M cysteamine. When cumulus-oocyte complexes (COCs) were matured in the mTCM-199 media supplemented with cysteamine, the rates of GVBD and maturation (metaphase II) were enhanced as compared to the media without the addition of cysteamine. Also, when COCs were matured in the mTCM-199 media supplemented with cysteamine, the rates of sperm penetration, male pronucleus formation, cleavage and blastocyst formation after in vitro fertilization were enhanced as compared to the media without the addition of cysteamine. In conclusion, it was suggested that oocytes matured for the first 22 h in mTCM-199 media supplemented with 150 $\mu$M cysteamine increased the rates of metaphase II, sperm penetration, male pronucleus and blastocyst formation were higher as compared to the media without addition of cysteamine.

Keywords

References

  1. Bing, Y. Z., Nagai, T. and Rodriguez-Martinez, H. 2001. Effect of cysteamine, FSH and estradiol-17$\beta$ on in vitro maturation of porcine oocyte. Theriogenology 55(4):867-876
  2. Calvin, H. I., Grosshans, K. and Blake, E. J. 1986. Estimation and manipulation of glutathione levels in prepuberal mouse ovaries and ova: relevance to sperm nucleus transformation in the fertilized egg. Gamete Res. 14:265-275 https://doi.org/10.1002/mrd.1120140310
  3. de Matos, D. G., Furnus, C. C., Moses, D. F., Martinez, A. G. and Matkovic, M. 1996. Stimulation of glutathione synthesis of in vitro matured bovine oocytes and its effect on embryo development and freezability. Mol. Reprod. Dev. 45:451-457 https://doi.org/10.1002/(SICI)1098-2795(199612)45:4<451::AID-MRD7>3.0.CO;2-Q
  4. Funahashi, H., Stumpf, T. T., Kim, N. H. and Day, B. N. 1996. Low salt maturation medium enhances histone HI kinase activity of porcine oocytes at the end of in vitro maturation. J. Reprod. Dev. 42:109-115 https://doi.org/10.1262/jrd.42.109
  5. Grupen, C. G., Nagashima, H. and Nottle, M. B. 1995. Cysteamine enhances in vitro development of porcine oocytes matured and fertilized in vitro. BioI. Reprod. 53:173-178 https://doi.org/10.1095/biolreprod53.1.173
  6. Ishii, T., Bannai, S. and Sugita, Y. 1981. Mechanism of growth stimulation of L1210 cells by 2-mercaptoethannol in vitro. J. BioI. Chem. 256:12387-12392
  7. Isseles, R. D., Nagele, A., Eckert, K. G. and Wilmanns, W. 1988. Promotion of cysteine uptake and its utilization for glutathione biosynthesis induced by cysteamine and N-acetylcy-steine. Biochem. Pharmacol. 37:881-888 https://doi.org/10.1016/0006-2952(88)90176-1
  8. Miyamura, M., Yoshida, M., Hamano, S. and Kuwayama, M. 1995. Glutathione concentration. during maturation and fertilization in bovine oocytes. Theriogenology 43:282
  9. Nagai, T. 1996. In vitro maturation and fertilization of pig oocytes. Anim. Reprod. Sci. 42:153-163 https://doi.org/10.1016/0378-4320(96)01487-X
  10. Naito, K., Daen, F. P. and Toyoda, Y. 1992. Comparison of histone HI kinase activity during meiotic maturation between two types of porcine oocytes matured in different media in vitro. BioI. Reprod. 47:43-47
  11. Perreault, S. D., Wolff, R. A. and Zirkin, B. R. 1984. The role of disulfide bond reduction during mammalian sperm nuclear decondensation in vivo. Dev. BioI. 101:160-167 https://doi.org/10.1016/0012-1606(84)90126-X
  12. Perreault, S. D., Babee, R. R. and Slott, V. I. 1988. Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes. Dev. BioI. 125:181-186 https://doi.org/10.1016/0012-1606(88)90070-X
  13. SAS Institute Inc. 1996. SAS/STAT guide for personal computer 6.12. SAS Inst. Inc., Cary, NC., USA
  14. Takahashi, M., Nagai, T., Hamano, S., Kuwayama, M., Okamura, N. and Okano, A. 1993 Effect of thiol compounds on in vitro development and intracellular glutathione content of bovine embryos. BioI. Reprod. 49: 228-232
  15. Yoshida, M., Ishigaki, K. and Pursel, V. G. 1992. Effect of maturation media on male pronucleus formation in pig oocytes matured in vitro. Mol. Reprod. Dev., 31:68-71 https://doi.org/10.1002/mrd.1080310112
  16. Yoshida, M. 1993a. Role of glutathione in the maturation and fertilization of pig oocytes in vitro. Mol. Reprod. Dev. 35:76-81
  17. Yoshida, M., Ishigaki, K., Nagai, T., Chikyu, M. and Pursel, V. G. 1993b. Glutathione concentration during maturation and after fertilization in pig oocytes: relevance to the ability of oocytes to form male pronucleus. BioI. Reprod. 49:89-94
  18. Yamauchi, N. and Nagai, T. 1999. Male pronuclear formation in denuded porcine oocytes after in vitro maturation in the presence of cysteamine. BioI. Reprod. 61:828-833 https://doi.org/10.1095/biolreprod61.3.828
  19. Zmuda, J. and Friedenson, B. 1983. Changes in intracellular glutathione levels in stimulated and unstimulated lymphocytes in the presence of 2-mercaptoethanol and cysteine. J. Immunol. 130:362-364