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Castro LS, Hamilton TR, Mendes CM, et al. Sperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different stages of cryopreservation. J Anim Sci Biotechnol. 2016;7:17doi: 10.1186/s40104-016-0076-x.
Castro, L. S., Hamilton, T. R., Mendes, C. M., Nichi, M., Barnabe, V. H., Visintin, J. A., & Assumpção, M. E. (2016). Sperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different stages of cryopreservation. Journal of animal science and biotechnology, 717. https://doi.org/10.1186/s40104-016-0076-x
Castro, L S, et al. "Sperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different stages of cryopreservation." Journal of animal science and biotechnology vol. 7 (2016): 17. doi: https://doi.org/10.1186/s40104-016-0076-x
Castro LS, Hamilton TR, Mendes CM, Nichi M, Barnabe VH, Visintin JA, Assumpção ME. Sperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different stages of cryopreservation. J Anim Sci Biotechnol. 2016 Mar 05;7:17. doi: 10.1186/s40104-016-0076-x. eCollection 2016. PMID: 26949533; PMCID: PMC4779270.
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J Anim Sci Biotechnol. 2016 Mar 05;7:17. doi: 10.1186/s40104-016-0076-x. eCollection 2016.

Sperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different stages of cryopreservation.

Journal of animal science and biotechnology

L S Castro, T R S Hamilton, C M Mendes, M Nichi, V H Barnabe, J A Visintin, M E O A Assumpção

Affiliations

  1. Laboratory of Spermatozoa Biology, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.
  2. Laboratory of Spermatozoa Biology, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil ; Laboratory of In Vitro Fertilization, Cloning and Animal Transgenesis, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.
  3. Laboratory of Andrology. Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.

PMID: 26949533 PMCID: PMC4779270 DOI: 10.1186/s40104-016-0076-x

Abstract

BACKGROUND: In order to improve the efficiency of bovine sperm cryopreservation process, it is important to understand how spermatozoa respond to differences in temperature as well as the ability to recover its own metabolism. The combination between flow cytometry approach and antioxidant enzymes activity allows a more sensible evaluation of sperm cell during cryopreservation. The aim of this study was to evaluate sperm attributes and antioxidant enzymes activity during different stages of cryopreservation process. Semen samples from Holstein bulls (n = 4) were separated in 3 treatments: fresh (37 °C); cooled (5 °C); and thawed. Evaluation occurred at 0 h and 2 h after incubation. Membrane integrity, mitochondrial membrane potential (MMP) and DNA damages were evaluated by flow cytometry; activities of antioxidant enzymes such as catalase, superoxide dismutase and gluthatione peroxidase were measured by spectrofotometry.

RESULTS: There was an increase in the percentage of sperm with DNA damage in the thawed group, compared to fresh and cooled, and for 2 hs of incubation when compared to 0 h. Considering MMP, there was an increase in the percentage of cells with medium potential in thawed group when compared to fresh and cooled groups. Opposingly, a decrease was observed in the thawed group considering high mitochondrial potential. Also in the thawed group, there was an increase on cells with damaged acrosome and membrane when compared to fresh and cooled groups. Significant correlations were found between antioxidant enzymes activity and membrane or mitochondrial parameters.

CONCLUSION: Based on our results, we conclude that cryopreservation affects cellular and DNA integrity and that the critical moment is when sperm cells are exposed to freezing temperature. Also, our study indicates that intracellular antioxidant machinery (SOD and GPX enzymes) is not enough to control cryodamage.

Keywords: Bovine; DNA integrity; JC-1; Sperm viability

References

  1. Hum Reprod. 2009 Sep;24(9):2061-70 - PubMed
  2. Anim Reprod Sci. 1999 May 17;56(1):51-65 - PubMed
  3. Theriogenology. 2006 Sep 1;66(4):822-8 - PubMed
  4. Anim Reprod Sci. 2012 Oct;134(3-4):184-90 - PubMed
  5. Biol Reprod. 2004 Sep;71(3):973-8 - PubMed
  6. Annu Rev Pharmacol Toxicol. 2007;47:143-83 - PubMed
  7. Science. 1999 Aug 27;285(5432):1393-6 - PubMed
  8. Biol Reprod. 1980 May;22(4):965-9 - PubMed
  9. J Clin Endocrinol Metab. 2008 Aug;93(8):3199-207 - PubMed
  10. J Assist Reprod Genet. 2013 Apr;30(4):461-77 - PubMed
  11. Fertil Steril. 2010 Mar 1;93(4):1027-36 - PubMed
  12. Oxid Med Cell Longev. 2016;2016:8213071 - PubMed
  13. Int J Androl. 2004 Apr;27(2):108-14 - PubMed
  14. Theriogenology. 2007 Sep 15;68(5):745-54 - PubMed
  15. Plant Physiol. 1970 Aug;46(2):200-3 - PubMed
  16. Anim Reprod Sci. 2010 Jan;117(1-2):34-42 - PubMed
  17. Theriogenology. 2001 Sep 1;56(4):557-67 - PubMed
  18. J Androl. 2008 Mar-Apr;29(2):213-21 - PubMed
  19. ILAR J. 2000;41(4):187-96 - PubMed
  20. Theriogenology. 2003 Sep 1;60(4):743-58 - PubMed
  21. Am J Physiol Heart Circ Physiol. 2007 May;292(5):H2023-31 - PubMed
  22. Hum Reprod. 2000 Oct;15(10):2160-4 - PubMed
  23. Reproduction. 2013 Oct 01;146(5):433-41 - PubMed
  24. Reproduction. 2013 Oct 01;146(5):R163-74 - PubMed
  25. J Androl. 2002 Mar-Apr;23(2):259-69 - PubMed
  26. Theriogenology. 2012 May;77(8):1497-504 - PubMed
  27. Oxid Med Cell Longev. 2016;2016:1687657 - PubMed
  28. Anim Reprod Sci. 2012 Dec;136(1-2):33-41 - PubMed
  29. J Androl. 1990 Jan-Feb;11(1):73-88 - PubMed
  30. Int J Androl. 2006 Feb;29(1):69-75; discussion 105-8 - PubMed
  31. J Androl. 2006 Mar-Apr;27(2):176-88 - PubMed
  32. J Androl. 2008 Jul-Aug;29(4):459-67 - PubMed
  33. Cryobiology. 2008 Feb;56(1):100-2 - PubMed
  34. Theriogenology. 2012 Sep 15;78(5):1005-19 - PubMed
  35. J Androl. 2000 Jan-Feb;21(1):1-7 - PubMed
  36. J Exp Zool. 1999 Nov 1;284(6):696-704 - PubMed
  37. Anim Reprod Sci. 2004 Feb;80(3-4):225-35 - PubMed
  38. Cryobiology. 1986 Feb;23(1):64-71 - PubMed
  39. Reprod Fertil Dev. 1993;5(6):639-58 - PubMed
  40. Mol Reprod Dev. 2000 Mar;55(3):282-8 - PubMed
  41. Methods Cell Sci. 2000;22(2-3):169-89 - PubMed
  42. Theriogenology. 2003 Jan 15;59(2):571-84 - PubMed
  43. Fertil Steril. 2001 Feb;75(2):263-8 - PubMed
  44. Reproduction. 2011 Dec;142(6):759-78 - PubMed
  45. Int J Androl. 1999 Jun;22(3):155-62 - PubMed
  46. Eur J Obstet Gynecol Reprod Biol. 1996 Jan;64(1):115-8 - PubMed
  47. J Exp Zool. 1993 Mar 15;265(4):432-7 - PubMed
  48. Free Radic Biol Med. 2006 Aug 15;41(4):528-40 - PubMed
  49. Reprod Fertil Dev. 2006;18(7):781-8 - PubMed
  50. Theriogenology. 2011 Jun;75(9):1623-9 - PubMed
  51. Anim Reprod Sci. 2000 Jul 2;60-61:481-92 - PubMed

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