Relation between Bull Sperm Respiratory Burst Activity and the In Vitro Fertilization Rate: A New Approach to Evaluate Bull's Fertility

Abstract

Sperm of 8 different bulls (3 ejaculates per bull) was used in the present study to evaluate the oxidative burst activity of each individual sperm cell by flow cytometry, correlating this data with the results of in vitro fertilization as well as the further embryo development to the stage of blastocyst. After thawing, the straw content was split in two identical parts. One was employed for the in vitro fertilization, while the other was used for flow cytometry to evaluate sperm oxidative burst activity by an assay using 2′,7′-dichlorofluorescin diacetate, which based on the oxidation of nonfluorescent DCFH-DA to highly fluorescent 2′,7′-dichlorofluorescein (DCF) intracellularly during the respiratory burst. As far as embryo production and the sperm metabolism oxidative burst activity is concerned, it was observed that bulls, in which the burst activity was higher, resulted in better results of in vitro fertilization and on the further embryo production. The correlation between burst activity and fertilization rate and further embryo development to blastocysts was, respectively, 95.6% and 87.8% (P≤.01). This study allows concluding that there is a positive correlation between bull sperm H2O2 production and their ability to fertilize bovine oocytes as well as their development to the blastocyst stage: sperm with high H2O2 production have high metabolic activities and consequently higher fertilizing capacities.

References

1. R. S. Jeyendran and E. F. Graham, “An evaluation of cryoprotective compounds on bovine spermatozoa,” Cryobiology, vol. 17, no. 5, pp. 458–464, 1980. View at: Google Scholar
2. A. Januskauskas, M. G. Håård, M. C. Håård, L. Söderquist, N. Lundeheim, and H. Rodriguez-Martinez, “Estimation of sperm viability in frozen-thawed semen from Swedish A.I. bulls,” Zentralblatt für Veterinärmedizin. Reihe A, vol. 43, no. 5, pp. 281–287, 1996. View at: Google Scholar
3. H. Kjaestad, E. Ropstad, and K. A. Berg, “Evaluation of spermatological parameters used to predict the fertility of frozen bull semen,” Acta Veterinaria Scandinavica, vol. 34, no. 3, pp. 299–303, 1993. View at: Google Scholar
4. R. P. Amann and R. H. Hammerstedt, “Andrology lab corner: detection of differences in fertility,” Journal of Andrology, vol. 23, no. 3, pp. 317–325, 2002. View at: Google Scholar
5. H. Rodríguez-Martínez, “Laboratory semen assessment and prediction of fertility: still utopia?,” Reproduction in Domestic Animals, vol. 38, no. 4, pp. 312–318, 2003. View at: Publisher Site | Google Scholar
6. P. R. Budworth, R. P. Amann, and P. L. Chapman, “Relationships between computerized measurements of motion of frozen-thawed bull spermatozoa and fertility,” Journal of Andrology, vol. 9, no. 1, pp. 41–54, 1988. View at: Google Scholar
7. C. Holt, W. V. Holt, H. D. M. Moore, H. C. B. Reed, and R. M. Curnock, “Objectivily measured boar sperm motility parameters correlate with the outcomes of on-farm inseminations: results of two fertility trials,” Journal of Andrology, vol. 18, no. 3, pp. 312–323, 1997. View at: Google Scholar
8. B. R. Zhang, B. Larsson, N. Lundeheim, and H. Rodriguez-Martinez, “Sperm characteristics and zona pellucida binding in relation to field fertility of frozen-thawed semen from dairy AI bulls,” International Journal of Andrology, vol. 21, no. 4, pp. 207–216, 1998. View at: Publisher Site | Google Scholar
9. A. Chaveiro, P. Santos, and F. M. da Silva, “Assessment of sperm apoptosis in cryopreserved bull semen after swim-up treatment: a flow cytometric study,” Reproduction in Domestic Animals, vol. 42, no. 1, pp. 17–21, 2007. View at: Publisher Site | Google Scholar
10. G. Antunes, A. Chaveiro, P. Santos, A. Marques, H. S. Jin, and F. Moreira da Silva, “Influence of apoptosis in bovine embryo's development,” Reproduction in Domestic Animals. Epub ahead of print. View at: Publisher Site | Google Scholar
11. F.-J. Liu, G. P. Cobb, T. A. Anderson, Q.-Q. Cheng, and C. W. Theodorakis, “Uptake, accumulation and depuration of sodium perchlorate and sodium arsenate in zebrafish (Danio rerio),” Chemosphere, vol. 65, no. 10, pp. 1679–1689, 2006. View at: Publisher Site | Google Scholar
12. R. Kasimanickam, K. D. Pelzer, V. Kasimanickam, W. S. Swecker, and C. D. Thatcher, “Association of classical semen parameters, sperm DNA fragmentation index, lipid peroxidation and antioxidant enzymatic activity of semen in ram-lambs,” Theriogenology, vol. 65, no. 7, pp. 1407–1421, 2006. View at: Publisher Site | Google Scholar
13. F. Moreira da Silva, Studies on reproduction and immunity: effect of the reproductive hotmones on the oxidative burst activity of bovine neutrophils, Ph.D. thesis, University of Gent, Gent, Belgium, 1997.
14. B. Halliwell, “Reactive oxygen species in living systems: source, biochemistry, and role in human disease,” American Journal of Medicine, vol. 91, no. 3, pp. 14S–22S, 1991. View at: Google Scholar
15. S. Raha and B. H. Robinson, “Mitochondria, oxygen free radicals, disease and ageing,” Trends in Biochemical Sciences, vol. 25, no. 10, pp. 502–508, 2000. View at: Publisher Site | Google Scholar
16. A. D. N. J. de Grey, “A hypothesis for the minimal overall structure of the mammalian plasma membrane redox system,” Protoplasma, vol. 221, no. 1-2, pp. 3–9, 2003. View at: Publisher Site | Google Scholar
17. J. F. H. M. Brouwers and B. M. Gadella, “In situ detection and localization of lipid peroxidation in individual bovine sperm cells,” Free Radical Biology and Medicine, vol. 35, no. 11, pp. 1382–1391, 2003. View at: Publisher Site | Google Scholar
18. B. A. Ball, A. T. Vo, and J. Baumber, “Generation of reactive oxygen species by equine spermatozoa,” American Journal of Veterinary Research, vol. 62, no. 4, pp. 508–515, 2001. View at: Google Scholar
19. B. Bánfi, G. Molnár, A. Maturana et al., “A ${\text{Ca}}^{2+}$-activated NADPH oxidase in testis, spleen, and lymph nodes,” The Journal of Biological Chemistry, vol. 276, no. 40, pp. 37594–37601, 2001. View at: Publisher Site | Google Scholar
20. J. McLeod, “The role of oxygen in the metabolism and motility of human spermatozoa,” American Journal of Physiology, vol. 138, pp. 512–518, 1943. View at: Google Scholar
21. R. J. Aitken and J. S. Clarkson, “Cellular basis of defective sperm function and its association with the genesis of reactive oxygen species by human spermatozoa,” Journal of Reproduction and Fertility, vol. 81, no. 2, pp. 459–469, 1987. View at: Google Scholar
22. R. J. Aitken, D. Buckingham, and D. Harkiss, “Use of a xanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa,” Journal of Reproduction and Fertility, vol. 97, no. 2, pp. 441–450, 1993. View at: Google Scholar
23. J. F. Griveau, P. Renard, and D. Le Lannou, “An in vitro promoting role for hydrogen peroxide in human sperm capacitation,” International Journal of Andrology, vol. 17, no. 6, pp. 300–307, 1994. View at: Google Scholar
24. I. Bize, G. Santander, P. Cabello, D. Driscoll, and C. Sharpe, “Hydrogen peroxide is involved in hamster sperm capacitation in vitro,” Biology of Reproduction, vol. 44, no. 3, pp. 398–403, 1991. View at: Google Scholar
25. C. O'Flaherty, N. Beorlegui, and M. Beconi, “Role of superoxide anion and hydrogen peroxide in acrosome reaction of bovine spermatozoa,” in Andrology in the 21st Century, B. Robaire, H. Chemes, and C. Morales, Eds., pp. 103–108, 2001. View at: Google Scholar
26. E. de Lamirande, H. Jiang, A. Zini, H. Kodama, and C. Gagnon, “Reactive oxygen species and sperm physiology,” Reviews of Reproduction, vol. 2, no. 1, pp. 48–54, 1997. View at: Publisher Site | Google Scholar
27. P. Leclerc, E. de Lamirande, and C. Gagnon, “Regulation of protein-tyrosine phosphorylation and human sperm capacitation by reactive oxygen derivatives,” Free Radical Biology and Medicine, vol. 22, no. 4, pp. 643–656, 1997. View at: Publisher Site | Google Scholar
28. A. Agarwal and T. M. Said, “Oxidative stress, DNA damage and apoptosis in male infertility: a clinical approach,” BJU International, vol. 95, no. 4, pp. 503–507, 2005. View at: Publisher Site | Google Scholar
29. M. Shekarriz, A. J. Thomas, Jr., and A. Agarwal, “Incidence and level of seminal reactive oxygen species in normal men,” Urology, vol. 45, no. 1, pp. 103–107, 1995. View at: Publisher Site | Google Scholar
30. S. E. M. Lewis, E. S. L. Sterling, I. S. Young, and W. Thompson, “Comparison of individual antioxidants of sperm and seminal plasma in fertile and infertile men,” Fertility and Sterility, vol. 67, no. 1, pp. 142–147, 1997. View at: Publisher Site | Google Scholar
31. S. Klebanoff and R. Clark, “The metabolic burst,” in The Neutrophil: Function and Clinical Disorders, S. J. Klebanoff and R. A. Clark, Eds., pp. 283–408, Elsevier/North-Holland Biomedical Press, Amsterdam, The Netherlands, 1978. View at: Google Scholar
32. D. A. Bass, J. W. Parce, L. R. Dechatelet, P. Szejda, M. C. Seeds, and M. Thomas, “Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation,” Journal of Immunology, vol. 130, no. 4, pp. 1910–1917, 1983. View at: Google Scholar
33. K. Whittington and W. C. L. Ford, “Relative contribution of leukocytes and of spermatozoa so reactive oxygen species production in human sperm suspensions,” International Journal of Andrology, vol. 22, no. 4, pp. 229–235, 1999. View at: Publisher Site | Google Scholar
34. C. P. LeBel, H. Ischiropoulos, and S. C. Bondy, “Evaluation of the probe $2^{\prime },7^{\prime }$-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress,” Chemical Research in Toxicology, vol. 5, no. 2, pp. 227–231, 1992. View at: Google Scholar
35. A. Chaveiro and F. Moreira da Silva, “Effect of oestrous cycle on the oxidative burst activity of blood polymorphonuclear leucocytes in cows,” Reproduction in Domestic Animals. Epub ahead of print. View at: Publisher Site | Google Scholar

Copyright

Copyright © 2010 António Chaveiro et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.