Effects of Pyruvate and Taurine for In Vitro Preservation in Boar Semen and the Developmental Rates of Embryos Fertilized by Semen Treated with Antioxidant

  • Jang H. Y. (College of Animal Resource Science, Kangwon National University) ;
  • Cheong H. T. (College of Animal Resource Science, Kangwon National University) ;
  • Kim C. I. (College of Animal Resource Science, Kangwon National University) ;
  • Park C. K. (College of Animal Resource Science, Kangwon National University) ;
  • Yang B. K. (Division of Animal Resource, Kangwon National University)
  • Published : 2005.06.01


Oxidative stress is one of the major causes of failure in in vitro storage of boar semen. Reactive oxygen species (ROS) are known to be important mediators of such stress. The present study examined the effects of pyruvate and taurine on sperm motility and expression of BAD, Cytochrome c, Caspase-3 and Cox-2 protein in in vitro storage of boar semen, and tested the effect of semen treated with antioxidant with or without hydrogen peroxide on the development of IVM/IVF porcine embryos. Semen samples were transported to the laboratory at $17^{\circ}C$ within 2 hr after collection and were treated with different concentration of pyruvate $(1\~10mM)$ and taurine $(25\~100mM)$ with or without 250uM $H_2O_2$ respectively. The supplementation of pyruvate and taurine increased sperm motility in boar semen during in vitro incubation at $37^{\circ}C$. Expression of apoptosis protein (BAD, cytochrome c, caspase-3 and cox-2) were reduced in the group of boar semen treated with pyruvate and taurine when compared to the other groups. The developmental rates of IVM/IVF porcine embryos fertilized by semen treated with pyruvate and taurine were significantly increased when compared to control (P<0.005). These results indicate that supplementation of pyruvate and taurine as antioxidants in boar semen extender can improve the semen quality and increase in vitro development of porcine IVM/IVF embryos when boar semen treated with antioxidants was used for in vitro fertilization.


  1. Abeydeera LR, Day BN (1997): Fertilization and subsequent development in vitro of pig ooeytes inseminated in a modified tris-buffered medium with frozen-thawed ejaculated spermatozoa. Biol Reprod 57(4):729-34 https://doi.org/10.1095/biolreprod57.4.729
  2. Aitken JR, Clarkson JS, Fishel S (1989): Generation of reactive oxygen species, lipid peroxidation and human sperm function. Biol Reprod 40:183-197
  3. Alexopoulos C, Boscos C, Saratsis PH, Saoulidis C, Kyriakis S (1996): The effect of storage time and number of spermatozoa per insemination dose on semen characteristics and fertilizing capacity of boar semen diluted with Beltsville Thaw Solution (BTS) extender. Anim Sci 62:599-604 https://doi.org/10.1017/S1357729800015150
  4. Cerolini S, Maldjian A, Surai P, Noble R (2000): Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Anim Reprod Sci 58(1-2):99-111 https://doi.org/10.1016/S0378-4320(99)00035-4
  5. Chen CS, Chao HI, Pan RL, Wei YH (1997): Hydroxyl radical-induced decline in motility and increase in lipid peroxidation and DNA modification in human sperm. Biochem Mol Biol Int 43:291-303
  6. De Lamirande E, Gagnon C (1994): Reactive oxygen species (ROS) and reproduction. Adv Exp Med Biol 366:185-97
  7. Fraga GG, Motchnik P A, Shigenaga MK, Helbrock JG, Jacob RA, Ames B (1991): Ascorbic acid protects against endogenous oxidative DNA damage in human sperm. Proc Natl Acad Sci USA 88:11003-110
  8. Halliwell B, Aruoma OI (1991): DNA damage by oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS Lett 281:9-19 https://doi.org/10.1016/0014-5793(91)80347-6
  9. Jang BY, Kong HS, Lee SS, Choi KD, Jeon GJ, Yang BK, Lee HK (2004): Expression of the antioxidant enzyme and apoptosis genes in in vitro maturation /in vitro fertilization porcine embryos. Asian-Aust. J Anim Sci 17: 33-38
  10. Korniewicz D, Fabianczyk B, Smorag Z (1996): The survival rate and fertilizing capacity of boar semen diluted with different diluents: III. International conference on boar semen preservation. Reprod Domest Anim 311:273-274
  11. Machaty Z, Takas T, Gathy, I (1992): Fertilizing capacity of boar semen diluted with Beltsville TS, BTS and Modified Kiev MK extenders in relation to storage time and number of spermatozoa per insemination dose. Anim Reprod Sci 29:289-295 https://doi.org/10.1016/0378-4320(92)90041-B
  12. O'Donnell Tormey J, Nathan CF, Lanks K (1987): Secretion of pyruvate: an antioxidant defense of mammalian cells. J Exp Med 165:500-514 https://doi.org/10.1084/jem.165.2.500
  13. Ogasawara M, Nakamura T, Koyama I (1993): Reactivity of taurine with aldehydes and its physiological role. Chem Pharm Bull 41:2172-2175 https://doi.org/10.1248/cpb.41.2172
  14. Sakkas D, Marietho E, Manicardi G (1999): Origin of DNA damage in ejaculated human spermatozoa. Rev Reprod 4:31-7 https://doi.org/10.1530/ror.0.0040031
  15. Sakkas D, Moffatt O, Manicardi GC, Mariethoz E, Tarozzi N, Bizzaro D (2002): Nature of DNA damage in ejaculated spermatozoa and the possible involvement of apoptosis. Biol Reprod 66:1061-7 https://doi.org/10.1095/biolreprod66.4.1061
  16. Stubbs CD, Smith AD (1984): The modification of mammalian membrane polyunsaturated fatty acid composition in relation to membrane fluidity and function. Biochim. Biophys Acta 779(1):89-137 https://doi.org/10.1016/0304-4157(84)90005-4
  17. Van Winkle LJ, Dickinson HR (1995): Differences in amino acid content of preimplantation mouse embryos that develop in vitro vs in vivo. In vitro effects of 5 amino acids that are abundant in oviductal secretions. Biol Reprod 52:96-104 https://doi.org/10.1095/biolreprod52.1.96
  18. Vermes I, Haanen C, Steffens-Nakken H, Reutenling-sperger C (1995): A novel assay for apoptosis. Flow cytometric detection of phosaphatidyl serine expression on early apoptotic ells using fluorescein labelled annexin V. J Immunol Methods 184:39-51 https://doi.org/10.1016/0022-1759(95)00072-I
  19. Wang S, Sharma RK, Ranganathan P, Thomas AJ, Falcone Jr T, Agarwal A (2001): Oxidative stress and increased levels of apoptosis (cytochrome C, Caspase 3 and 9) in patients with male-factor infertility. Fertil Steril 76:S195
  20. Weng SL, Taylor SL, Morshedi M, Schuffner A (2002): Caspase activity and apoptotic markers in ejaculated human sperm. Mol Hum Reprod 8:984-91 https://doi.org/10.1093/molehr/8.11.984
  21. Yang J, Liu X, Bhalla K, Kim CN, Ibardo AM, Cai J (1997): Prevention of apoptosis by Bcl-2; release of Cytochrome c from mitochondria blocked. Science 275:1129-32 https://doi.org/10.1126/science.275.5303.1129