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Loss of Intracellular Antioxidant Enzyme Activity During Sperm Cryopreservation: Effects on Sperm Function After Thawing

  • Juan G. AlvarezEmail author
Chapter

Abstract

Despite advances in cryopreservation methodology, one of the main detrimental effects of cryopreservation on human spermatozoa is a marked reduction in motility. The primary cause of cellular damage during cryopreservation is the formation of intracellular ice. The concentration of solutes remaining in the unfrozen fraction increases, thereby both depressing the freezing point and increasing the osmotic pressure of the remaining solution. Hence, biological systems freeze progressively over a wide temperature range, during which the solute becomes gradually more concentrated as the temperature falls. This leads to irreversible rupturing of plasma and nuclear membranes and disturbance of cellular organelles. The nucleus has generally been considered to be a stable constituent of the cell. However, recent studies have suggested that this is not the case and that inappropriate chromatin condensation can occur with freezing. Cryoprotectants such as glycerol or propanediol can be added to cells to reduce freezing damage by lowering the salt concentrations and increasing the unfrozen water fraction, thereby reducing osmotic stress. Further cellular damage may be caused during the thawing process as the ice melts or recrystallizes. Slow thawing is most likely to induce injury, as it allows time for consolidation of microscopic ice crystals into larger forms which are known to be damaging. The production and dissolution of ice is associated with the actual rate of freezing and thawing. Slow freezing and gradual dehydration may accommodate cell survival, whereas rapid freezing and thawing is more likely to result in cell death.

Keywords

Sperm cryopreservation Sperm motility Lipid peroxidation Sperm function after thawing Oxygen radical production Antioxidant enzymes Superoxide dismutase Glutathione peroxidase DNA fragmentation DNA oxidation 

References

  1. 1.
    Centola GM, Raubertas RF, Mattox JH. Cryopreservation of human semen: comparison of cryopreservation, sources of variability, and prediction of post-thaw survival. J Androl. 1992;13:283–8.PubMedGoogle Scholar
  2. 2.
    Agarwal A, Tolentine MV, Sidhu Jr RS, et al. Effect of cryopreservation on semen quality in patients with testicular cancer. Urology. 1995;46:382–6.PubMedCrossRefGoogle Scholar
  3. 3.
    Critzer JK, Huse-Benda AR, Aaker DV, et al. Cryopreservation of human spermatozoa. III. The effects of cryoprotectants on motility. Fertil Steril. 1988;50:314–20.Google Scholar
  4. 4.
    Englert Y, Delvigne A, Vekemans M, et al. Is fresh or frozen semen to be used in in vitro fertilization with donor sperm. Fertil Steril. 1989;5:661–4.Google Scholar
  5. 5.
    Yoshida H, Hoshiai H, Fukaya T, et al. Fertilizability of fresh and frozen human spermatozoa. Assist Reprod Technol Androl. 1990;1:164–72.Google Scholar
  6. 6.
    Muldrew K, McGann LE. Mechanisms of intracellular ice formation. Bio J. 1990;57:525–33.Google Scholar
  7. 7.
    Watson PF. Recent developments and concepts in cryopreservation of spermatozoa and the assessment of their post-thaw function. Reprod Fertil Dev. 1995;7:871–91.PubMedCrossRefGoogle Scholar
  8. 8.
    Brotherton J. Cryopreservation of human semen. Arch Androl. 1990;25:181–95.PubMedCrossRefGoogle Scholar
  9. 9.
    Royere D, Hamamah S, Nicolle JC, et al. Freezing and thawing alter chromatin stability of ejaculated human spermatozoa: fluorescence acridine orange staining and fuelgen DNA cytophotometric studies. Gamete Res. 1988;21:51–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Royere D, Hamamah S, Nicolle JC, Lansac J. Chromatin alterations induced by freeze-thawing influence the fertilizing ability of human sperm. Int J Androl. 1991;14:328–32.PubMedCrossRefGoogle Scholar
  11. 11.
    Mazur P, Rall WF, Rigopoulos N. Relative contribution of the fraction of unfrozen water and of salt concentration to the survival of slowly frozen human erythrocytes. Biophys J. 1981;36:653–75.PubMedCrossRefGoogle Scholar
  12. 12.
    Chatterjee S, Gagnon C. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev. 2001;59:451–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Kalthur G, Adiga SK, Upadhya D, Rao S, Kumar P. Effect of cryopreservation on DNA integrity in patients with teratozoospermia. Fertil Steril. 2008;89:1723–7.PubMedCrossRefGoogle Scholar
  14. 14.
    Zribi N, Chakroun NF, El Euch H, Gargouri J, Bahloul A, Keskes LA. Effects of cryopreservation on human sperm deoxyribonucleic acid integrity. Fertil Steril. 2010;93:159–66.PubMedCrossRefGoogle Scholar
  15. 15.
    Gandini L, Lombardo F, Lenzi A, Spanò M, Dondero F. Cryopreservation and sperm DNA integrity. Cell Tissue Bank. 2006;7:91–8.PubMedCrossRefGoogle Scholar
  16. 16.
    Gosalvez J, Cortes-Gutierez E, Lopez-Fernandez C, Fernandez JL, Caballero P, Nunez R. Sperm deoxyribonucleic acid fragmentation dynamics in fertile donors. Fertil Steril. 2009 Jul;92(1):170–3.PubMedCrossRefGoogle Scholar
  17. 17.
    Toro E, Fernández S, Colomar A, Casanovas A, Alvarez JG, López-Teijón M, Velilla E. Processing of semen can result in increased sperm DNA fragmentation. Fertil Steril. 2009;92:2109–12.PubMedCrossRefGoogle Scholar
  18. 18.
    Aitken RJ. Free radicals, lipid peroxidation and sperm function. Reprod Fertil Dev. 1995;7:659–68.PubMedCrossRefGoogle Scholar
  19. 19.
    Calamera JC, Buffone MG, Doncel GF, Brugo-Olmedo S, de Vicentis S, Calamera MM, Storey BT, Alvarez JG. Effect of thawing temperature on the motility recovery of cryopreserved human spermatozoa. Fertil Steril. 2010;93:789–94.PubMedCrossRefGoogle Scholar
  20. 20.
    Ollero M, Powers RD, Alvarez JG. Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: implications for sperm lipoperoxidative damage. Mol Reprod Dev. 2000;55:326–34.PubMedCrossRefGoogle Scholar
  21. 21.
    Ollero M, Gil-Guzman E, Lopez MC, Sharma RK, Agarwal A, Larson K, Evenson D, Thomas Jr AJ, Alvarez JG. Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility. Hum Reprod. 2001;16:1912–21.PubMedCrossRefGoogle Scholar
  22. 22.
    Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ, Agarwal A. Differential production of reactive oxygen species by subsets of human spermatozoa at different stage of maturation. Hum Reprod. 2001;16:1022–30.CrossRefGoogle Scholar
  23. 23.
    Jones R, Mann T, Sherings RJ. Adverse effects of peroxidized lipid on human spermatozoa. Proc R Soc Lond B Biol Sci. 1978;201:413–7.PubMedCrossRefGoogle Scholar
  24. 24.
    Alvarez JG, Storey BT. Role of superoxide dismutase in protecting rabbit spermatozoa from O2 toxicity due to lipid peroxidation. Biol Reprod. 1983;28:1129–36.PubMedCrossRefGoogle Scholar
  25. 25.
    Alvarez JG, Storey BT. Assessment of cell damage caused by spontaneous lipid peroxidation in rabbit spermatozoa. Biol Reprod. 1984;30:323–32.PubMedCrossRefGoogle Scholar
  26. 26.
    Alvarez JG, Storey BT. Role of glutathione peroxidise in protecting mammalian spermatozoa from loss of motility caused by spontaneous lipid peroxidation. Gamete Res. 1989;23:77–90.PubMedCrossRefGoogle Scholar
  27. 27.
    Alvarez JG, Storey BT. Taurine, hypotaurine, epinephrine and albumin inhibit lipid peroxidation in rabbit spermatozoa and protect against loss of motility. Biol Reprod. 1983;29:548–55.PubMedCrossRefGoogle Scholar
  28. 28.
    Kovalski NN, de Lamirande E, Gagnon C. Reactive oxygen species generated by human neutrophils inhibit sperm motility: protective effect of seminal plasma and scavengers. Fertil Steril. 1992;58:809–15.PubMedGoogle Scholar
  29. 29.
    Li TK. The glutathione and thiol content of mammalian spermatozoa and seminal plasma. Biol Reprod. 1975;12:641–6.PubMedCrossRefGoogle Scholar
  30. 30.
    Menella MRF, Jones R. Properties of spermatozoal superoxide dismutase and lack of involvement of superoxides in metals ion-catalyzed lipid peroxidation reactions in semen. Biochem J. 1980;191:289–97.Google Scholar
  31. 31.
    Alvarez JG, Touchstone JC, Blasco L, Storey BT. Spontaneous lipid peroxidation and production of hydrogen peroxide and superoxide in human spermatozoa. Superoxide dismutase as major enzyme protectant against oxygen toxicity. J Androl. 1987;8:338–48.PubMedGoogle Scholar
  32. 32.
    Gavella M, Lipovac V, Vucic M, Rocic B. Relationship of sperm superoxide dismutase-like activity with other sperm-specific enzymes’ and experimentally induced lipid peroxidation in infertile men. Andrologia. 1996;28:223–9.PubMedCrossRefGoogle Scholar
  33. 33.
    Aitken RJ, Buckingham DW, Carreras A, Irvine DS. Superoxide dismutase in human sperm suspensions: relationship with cellular composition, oxidative stress, and sperm function. Free Radic Biol Med. 1996;21:495–504.PubMedCrossRefGoogle Scholar
  34. 34.
    Alvarez JG, Storey BT. Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation. J Androl. 1992;13:232–41.PubMedGoogle Scholar
  35. 35.
    Holland MK, Alvarez JG, Storey BT. Production of superoxide and activity of superoxide dismutase in rabbit epididymal spermatozoa. Biol Reprod. 1982;27:1109–18.PubMedCrossRefGoogle Scholar
  36. 36.
    Buffone MG, Doncel GF, Marín Briggiler CI, Vazquez-Levin MH, Calamera JC. Human sperm subpopulations: relationship between functional quality and protein tyrosine phosphorylation. Hum Reprod. 2004;19:139–46.PubMedCrossRefGoogle Scholar
  37. 37.
    Alvarez et al. Dynamics of DNA fragmentation after thawing of subsets of cryopreserved human spermatozoa. Annual Meeting of the Spanish Fertility Society. Valencia, Spain; 2010.Google Scholar
  38. 38.
    Bahçeci M, Ciray HN, Karagenc L, Uluğ U, Bener F. Effect of oxygen concentration during the incubation of embryos of women undergoing ICSI and embryo transfer: a prospective randomized study. Reprod Biomed Online. 2005;11:438–43.PubMedCrossRefGoogle Scholar
  39. 39.
    Meintjes M, Chantilis SJ, Douglas JD, et al. A controlled randomized trial evaluating the effect of lowered incubator oxygen tension on live births in a predominantly blastocyst transfer program. Hum Reprod. 2009;24:300–7.PubMedCrossRefGoogle Scholar
  40. 40.
    Kovačič B, Sajko MC, Vlaisavljević V. A prospective, randomized trial on the effect of atmospheric versus reduced oxygen concentration on the outcome of intracytoplasmic sperm injection cycles. Fertil Steril. 2010;94:511–9.PubMedCrossRefGoogle Scholar
  41. 41.
    Weksler-Zangen S, Yaffe P, Ornoy A. Reduced SOD activity and increase neural tube defects in embryos of the sensitive but not of the resistant Cohen diabetic rats cultured under aerobic conditions. Birth Defects Res A Clin Mol Teratol. 2003;67:429–37.PubMedCrossRefGoogle Scholar

Further Reading

  1. Aitken RJ. Evaluation of human sperm function. Br Med Bull. 1990;46:654–74.PubMedGoogle Scholar
  2. Alvarez JG, Storey BT. Differential incorporation of fatty acids into and peroxidative loss of fatty acids from phospholipids of human spermatozoa. Mol Reprod Dev. 1995;42:334–46.PubMedCrossRefGoogle Scholar
  3. Calamera JC, Brugo S, Vilar O. Relation between motility and adenosinetriphosphate (ATP) in human spermatozoa. Andrologia. 1982;14:239–41.PubMedCrossRefGoogle Scholar
  4. Calamera JC, Doncel GF, Olmedo SB, Kolm P, Acosta AA. “Modified sperm stress test: a simple assay that predicts sperm-related abnormal in-vitro fertilization. Hum Reprod. 1998;13:2484–8.PubMedCrossRefGoogle Scholar
  5. Gravance CG, Vishwanath R, Pitt C, Garner DL, Casey PJ. Effects of cryopreservation on bull sperm head morphometry. J Androl. 1998;19:704–9.PubMedGoogle Scholar
  6. Jones R, Mann T, Sherins R. Peroxidative breakdown of phospholipids in human spermatozoa, spermicidal properties of fatty acid peroxides, and protective action of seminal plasma. Fertil Steril. 1979;31:531–7.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Centro de Infertilidad Masculina AndrogenLa CoruñaSpain
  2. 2.Harvard Medical SchoolBostonUSA

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