Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Effects of Reactive Oxygen Species on the Function of Porcine Spermatozoa

활성산소계가 돼지 정자의 기능에 미치는 영향

  • Kim, Byung-Gak (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Ki-Jung (Department of Animal Science & Technology, Chung-Ang University) ;
  • Lee, Yong-An (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Bang-Jin (Department of Animal Science & Technology, Chung-Ang University) ;
  • Kim, Yong-Hee (Department of Animal Science & Technology, Chung-Ang University) ;
  • Ryu, Buom-Yong (Department of Animal Science & Technology, Chung-Ang University)
  • 김병각 (중앙대학교 동물자원과학과) ;
  • 김기중 (중앙대학교 동물자원과학과) ;
  • 이용안 (중앙대학교 동물자원과학과) ;
  • 김방진 (중앙대학교 동물자원과학과) ;
  • 김용희 (중앙대학교 동물자원과학과) ;
  • 류범용 (중앙대학교 동물자원과학과)
  • Published : 2009.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

The current study was designed to evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase system (XO) on sperm function and DNA fragmentation in porcine spermatozoa. ROS were produced by a combination of $1,000{\mu}M$ X and 50 mU/ml XO. The ROS scavengers such as superoxide dismutase (SOD) (200 U/ml) and catalase (CAT) (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without antioxidants at $37^{\circ}C$ under 5% $CO_2$ incubator. Ca-ionophore-induced acrosome reaction, the proportion of swollen spermatozoa under hypo-osmotic condition, malondialdehyde formation for the analysis of lipid peroxidation, and the proportion of DNA fragmentation were determined after 2 hours incubation. The action of ROS on porcine spermatozoa resulted in decreased Ca-ionophore-induced acrosome reaction and membrane integrity, increased the formation of malondialdehyde, and the proportion of sperm with DNA fragmentation(p<0.05). The toxic effects caused by ROS were completely alleviated by CAT in terms of sperm function and characteristics, however SOD did not serve the same scavenger effect as CAT. To conclude, the ROS can cause significant damage to porcine sperm functions and characteristics, which can be minimized by the use of antioxidants.

Keywords

References

  1. Aitken RJ, Clarkson JS, Hargreave TB, Irvine DS, Wu FC (1989): Analysis of the relationship between defective sperm function and the generation of reactive oxygen species in cases of oligozoospermia. J Androl 10:214-220 https://doi.org/10.1002/j.1939-4640.1989.tb00091.x
  2. Aitken RJ, West KM (1990): Analysis of the relationship between reactive oxygen species production and leucocyte infiltration in fractions of human semen separated on Percoll gradients. Int J Androl 13:433- 451 https://doi.org/10.1111/j.1365-2605.1990.tb01051.x
  3. Aitken RJ, Buckingham DW, Harkiss D (1993a): Use of a xanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa. J Reprod Fetil 97:441- 450 https://doi.org/10.1530/jrf.0.0970441
  4. Aitken RJ, Harkiss D, Buckingham DW (1993b): Analysis of lipid peroxidation mechanism in human spermatozoa. Mol Reprod Dev 35:302-315 https://doi.org/10.1002/mrd.1080350313
  5. Alvarez JG, Touchstone JC, Blasco L, Storey BT (1987): Spontaneous lipid peroxidation and production of hydrogen peroxide in human spermatozoa: superoxide dismutase as major enzyme protectant against oxygen toxicity. J Androl 8:338-348
  6. Balhorn R (1982): A model for the structure of chromatin in mammalian sperm. J Cell Biol 93:298-305 https://doi.org/10.1083/jcb.93.2.298
  7. Biggers JS, Whitten WK, Whittingham DG (1971): The culture of mouse embryos in vitro. In: Daniel JC, editor. Method in Mammalian Embryology. San Francisco, Freeman, pp 86-116
  8. Block ER (1991): Hydrogen peroxide alters the physical state and function of the plasma membrane of pulmonary artery endothelial cells. J Cell Physiol 20: 362-369
  9. Cummins JM, Pember SM, Jequier AM, Yovich JL (1991): A test of the human sperm acrosome reaction following ionophore challenge. J Androl 12:98-103
  10. de Lamirande E, Eiley D, Gagnon C (1993): Inverse relationship between the induction of human sperm capacitation and spontaneous acrosome reaction by various biological fluids and the superoxide scavenging capacity of these fluids. Int J Androl 16:258-266 https://doi.org/10.1111/j.1365-2605.1993.tb01189.x
  11. de Lamirande E, Gagnon C (1995): Impact of reactive oxygen species on spermatozoa; a balancing act between beneficial and detrimental effects. Hum Reprod 12 Suppl 1:15-21
  12. Griveau JF, Dumont E, Renard P, Callegari JP, Le Lannou D (1995): Reactive oxygen species, lipid peroxidation and enzymatic defence systems in human spermatozoa. J Reprod Fertil 103:17-26 https://doi.org/10.1530/jrf.0.1030017
  13. Guthrie HD, Welch GR (2006): Determination of intracellular reactive oxygen species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry. J Anim Sci 84:2089-2100 https://doi.org/10.2527/jas.2005-766
  14. Jeyendran RS, Van der Ven HH, Perez-Pelaez M, Crabo BG, Zaneveld LJ (1984): Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J Reprod Fertil 70:219-228 https://doi.org/10.1530/jrf.0.0700219
  15. Jones R, Mann T, Sherins RJ (1979): Peroxidative breakdown of phospholipids in human spermatozoa: spermicidal effects of fatty acid peroxides and protective action of seminal plasma. Fertil Steril 31:531- 537
  16. Mazzilli F, Rossi T, Marchesini M, Ronconi C, Dondero F (1994): Superoxide anion in human semen related to seminal parameters and clinical aspects. Fertil Steril 62:862-868
  17. Iwasaki A, Gagnon C (1992): Formation of reactive oxygen species in spermatozoa of infertility patients. Fertil Steril 57:2409-2416
  18. Lopes S, Jurisicova A, Sun JG, Casper RF (1998): Reactive oxygen species: potential cause for DNA fragmentation in human spermatozoa. Hum Reprod 13: 896-900 https://doi.org/10.1093/humrep/13.4.896
  19. Manicardi GC, Bianchi PG, Pantano S, Azzoni P, Bizzaro D, Bianchi U, Sakkas D (1995): Presence of endogenous nicks in DNA of ejaculated human spermatozoa and its relationship to chromomycin A3 accessibility. Biol Reprod 52:864-867 https://doi.org/10.1095/biolreprod52.4.864
  20. Ohyashiki T, Tsuka T, Mohri T (1988): Increase of the molecular rigidity of the protein conformation in the intestinal brush-border membranes by lipid peroxidation. Biochim Biophys Acta 939:383-392 https://doi.org/10.1016/0005-2736(88)90084-3
  21. Slater TF, Sawyer BC (1971): The stimulatory effects of carbon tetrachloride and other halogenoalkanes on peroxidative in rat liver fractions in vitro. Biochem J 123:805-814 https://doi.org/10.1042/bj1230805
  22. Smith R, Kaune H, Parodi D, Madariaga M, Rios R, Morales I, Castro A (2006): Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. Hum Reprod 21:986-993 https://doi.org/10.1093/humrep/dei429
  23. Twigg J, Fulton N, Gomez E, Irvine DS, Aitken RJ (1998): Analysis of the impact of intracellular reactive oxygen species generation on the structural and functional integrity of human spermatozoa: lipid peroxidation, DNA fragmentation and effectiveness of antioxidants. Hum Reprod 13:1429-1436 https://doi.org/10.1093/humrep/13.6.1429
  24. 류범용, 정영채, 김창근, 신현아, 한정호, 방명걸, 오선경, 김석현, 문신용 (2002): 소 정자에 있어서 활성산소계가 정자 기능과 지방산화 및 DNA 절편화에 미치는 영향. 대한불임학회지 29:105-115