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Relationship between nuclear chromatin decondensation (NCD) in vitro and other sperm parameters and their predictive value on fertilization rate in IVF program

Abstract

Purpose: The purpose of this study was to determine the in-vitro induced nuclear chromatin decondensation (NCD) of human spermatozoa and its value in combination with routine semen analysis in predicting the outcome of in-vitro fertilization (IVF).

Methods: The ejaculate of 52 couples, undergoing IVF, was incubated with lithium diidosalicylic acid (LIS) and dithiothreitol (DTT) (G.1) or with heparin and sodium dodecyl sulphate (SDS) (G.2) to induce chromatin decondensation (NCD). Smears were made at 30, 60, and 120 min after incubation.

Results: NCD was evaluated by morphometrical detection of the surface area of the spermatozoa using a semiautomatic image analysis system (IBAS). In both groups, the sperm heads showed a significant enlargement after 30, 60, and 120 min incubation in comparison to the initial size. However, no correlation was found between NCD at various periods of time and the fertilization rates. The mean area of the sperm heads in the native sample in the G.1 was 9.45± 1.33 μm2 and 9.02± 1.15 μm2 in the G.2. This area increased after incubation for 30, 60, and 120 min to 10.92± 1.48, 12.26± 2.16, 13.54± 3.14, and 15.35± 7.78 μm2 in the first group (G.1) and to 10.29± 1.15, 11.23± 1.85, 11.46± 1.97, and 11.27± 2.82 μm2 in the second group, respectively.

Conclusions: NCD in vitro after incubation with LIS + DTT or heparin + SDS could not be recommended as a predictive parameter for IVF outcome.

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References

  1. Bahlhorn R: A model for the structure of chromatin in mammalian sperm. J Cell Biol 1982;93:298–308

    Google Scholar 

  2. Bedford JM, Calvin HI: The occurrence and possible functional significance of –S–S– crosslink in sperm heads, with particular reference to eutherian mammals. J Exp Zool 1974;188:137–155

    Article  CAS  PubMed  Google Scholar 

  3. Perreault SD, Naish SJ, Zirkin BR, et al.: The timing of hamster sperm nuclear decondensation and male pronucleus formation is related to sperm nuclear disulfide bond contents. Biol Reprod 1987;36:239–244

    Article  CAS  PubMed  Google Scholar 

  4. Huret JL: Nuclear chromatin decondensation of human sperm: A review. Arch Androl 1986;16:97–109

    CAS  PubMed  Google Scholar 

  5. Calvin HI, Bedford JM: Stimulation of actinomycin-D-binding to eutherian sperm chromatin by reduction of disulphide bonds. J Reprod Fertil 1974;36:225–229

    CAS  PubMed  Google Scholar 

  6. Bjorndahl L, Kjellberg S, Roomans GM, Kvist U: The human sperm nucleus takes up zink at ejaculation. Int J Androl 1986;9:77–80

    CAS  PubMed  Google Scholar 

  7. Kvist U, Bjorndahl L, Kjellberg S: Sperm nuclear zinc, chromatin stability, and male fertility. Scanning Microsc 1987;1:1241–1247

    CAS  PubMed  Google Scholar 

  8. Rosenborg L, Rao KM, Bjorndal L, Kvist U, Pousette A, Ukerlψf E, Fredricson B: Changes in human sperm chromatin stability during preparation for in vitro fertilization. Int J Androl 1990;13:287–296

    CAS  PubMed  Google Scholar 

  9. Lipitz S, Bartoov B, Rajuan C. et al.: Sperm head ultramorphology and chromatin stability of males with unexplained infertility who fail to fertilize normal human ova in vitro. Andrologia 1992;24:261–269

    CAS  PubMed  Google Scholar 

  10. Matsuda Y, Yamada T, Tobari I: Studies on chromosomes abberation in the eggs of mice fertilized in vitro after irradiation. I. Chromosome aberration induced in sperm after X-irradiation. Mutat Res 1985;148:113–117

    CAS  PubMed  Google Scholar 

  11. Perreault SD, Wolff RA, Zirkin BR: The role of disulfide bond reduction during mammalian sperm nuclear decondensation in vivo. Dev Biol 1984;101:160–167

    Article  CAS  PubMed  Google Scholar 

  12. Yanagimachi R: Stability of the mammalian sperm nucleus. Zygote 1994;2:383–384

    CAS  PubMed  Google Scholar 

  13. Philpott A, Leno GH, Laskey RA: Sperm decondensation in Xenopus egg cytoplasma is mediated by nucleoplasmin. Cell 1991;65:569–578

    Article  CAS  PubMed  Google Scholar 

  14. Bedford JM, Kim HH: Sperm/egg binding patterns and oocytes cytology in retrospective analysis of fertilization failure in vitro. Hum Reprod 1993;8:453–463

    CAS  PubMed  Google Scholar 

  15. Zamboni L: Detection of human sperm pathology by fine structure analysis. In The Biological Basis of Early Human Reproductive Failure: Application to Medically Assisted Conception and the Treatment of Infertility. JV Blerkom (ed), New York, Oxford University Press, 1994, pp. 327–344

    Google Scholar 

  16. Lalich RA, Vedantham S, McCormick N, Wagner C, Prins GS: Relationship between heparin binding characteristics and ability of human spermatozoa to penetrate hamster ova. J Reprod Fertil 1989;86:297–302

    CAS  PubMed  Google Scholar 

  17. Kvist U, Eliasson R: Influence of seminal plasma on the chromatin stability of ejaculated human spermatozoa. J Androl 1980;1:130–142

    Google Scholar 

  18. Bedford JM: An electron microscopic study of sperm penetration into rabbit egg after natural mating. Am J Anat 1972;33:165–178

    Google Scholar 

  19. Soupart P, Stron PA: Ultrastructural observation on human oocytes fertilized in vitro. Fertil Steril 1974;25:11–44

    CAS  PubMed  Google Scholar 

  20. World Health Organization. Laboratory Manual for the Examination of Human Semen and Semen Cervical Mucus Interaction. Cambridge, UK, Cambridge University Press, 1999

    Google Scholar 

  21. Hammadeh ME, Kühnen A, Amer AS, Rosenbaum P, Schmidt W: Comparison of sperm preparation methods: Effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer. Int J Androl 2001;24:360–368

    CAS  PubMed  Google Scholar 

  22. Krüger TF, Menkfeld R, Stander FSH: Sperm morphologic feature as a prognostic factor in IVF. Fertil Steril 1986;46:118–123

    Google Scholar 

  23. Hammadeh ME, Al-Hasani S, Stieber M, et al.: The effect of chromatin condensation and morphology of human spermatozoa on fertilization, cleavage, and pregnancy rates in an intracytoplasmic sperm injection programme. Hum Reprod 1996;11:2468–2471

    CAS  PubMed  Google Scholar 

  24. Kvist U: Importance of spermatozoa zinc as temporary inhibitor of sperm nuclear chromatin decondensation ability in man. Acta Physiol Scand 1980;109:79–84

    CAS  PubMed  Google Scholar 

  25. Huret JL: Variability of the chromatin decondensation ability test on human sperm. Arch Androl 1983;11:1–7

    CAS  PubMed  Google Scholar 

  26. Mahi CA, Yanagimachi R: Induction of nuclear decondensation of mammalian spermatozoa in-vitro. J Reprod Fertil 1975;44:293–296

    CAS  PubMed  Google Scholar 

  27. Gall WE, Ohsumi Y: Decondensation of sperm nuclei in vitro. Exp Cell Res 1976;102:349–358

    Article  CAS  PubMed  Google Scholar 

  28. Delgado NM, Huacuja L, Merchant H, et al.: Species specific decondensation of human spermatozoa nuclei by heparin. Arch Androl 1980;8:87–95

    Google Scholar 

  29. Sakkas D, Urner F, Bianchi PG, et al.: Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection. Hum Reprod 1996;11:837–843

    CAS  PubMed  Google Scholar 

  30. Montag M, Tok V, Liow SL, et al.: In vitro decondensation of mammalian sperm and subsequent formation of pronuclei-like structures for micromanipulation. Mol Reprod Dev 1992;33:338–346

    Article  CAS  PubMed  Google Scholar 

  31. Banerjee S, Hulten MAJ: Sperm nuclear chromatin transformation in somatic cell—free extracts. Mol Reprod Dev 1994;37:305–317

    Article  CAS  PubMed  Google Scholar 

  32. Chitale AR, Rathaur RG: Nuclear decondensation of sperm head and failure at in vitro fertilization: An ultrastructure study. Hum Reprod 1995;10:594–598

    CAS  PubMed  Google Scholar 

  33. Samocha-Bon D, Klewin LM, Weissemberg R, et al.: In vitro human spermatozoa nuclear decondensation assessed by flow cytometry. Mol Hum Reprod 1998:4:133–137

    Google Scholar 

  34. Zucker RM, Perreault SD, Elstein KH: Utility of light scatter in the morphological analysis of sperm. Cytometry 1992;13:39–47

    Article  CAS  PubMed  Google Scholar 

  35. Evenson DP, Darzykiewicz Z, Melamed MR: Relation of mammalian sperm chromatin heterogeneity to fertility. Science 1980;240:1131–1133

    Google Scholar 

  36. Liu DY, Elton RA, Johnston WI, Baker HW: Spermatozoal nuclear chromatin decondensation in vitro: A test for sperm immaturity. Comparison with results of human in vitro fertilization. Clin Reprod Fertil 1987;5:191–201

    CAS  PubMed  Google Scholar 

  37. Gopalkrishnan K, Hinduja IN, Anand-Kumar TC: In vitro decondensation of nuclear chromatin of human spermatozoa: Assessing fertilizing potential. Arch Androl 1991;27:43–50

    CAS  PubMed  Google Scholar 

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Correspondence to Mohamed E. Hammadeh.

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Hammadeh, M.E., Bernardi, A., Zeginiadou, T. et al. Relationship between nuclear chromatin decondensation (NCD) in vitro and other sperm parameters and their predictive value on fertilization rate in IVF program. J Assist Reprod Genet 22, 301–305 (2005). https://doi.org/10.1007/s10815-005-6002-3

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  • DOI: https://doi.org/10.1007/s10815-005-6002-3

Key Words

  • Chromatin decondensation
  • human spermatozoa
  • image analysis
  • morphometry
  • surface area