Reproductive BioMedicine Online
Volume 19, Issue 5 , Pages 671-684 , November 2009

Chromomycin A3 staining, sperm chromatin structure assay and hyaluronic acid binding assay as predictors for assisted reproductive outcome

  • Martine Nijs

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • Corresponding Author InformationCorrespondence:
    • ,
  • Eva Creemers

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • ,
  • Annemie Cox

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • ,
  • Kim Franssen

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • ,
  • Mia Janssen

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • ,
  • Elke Vanheusden

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium
    • ,
  • Christopher De Jonge

      Affiliations

    • Reproductive Medicine Centre (RMC), University of Minnesota, Minneapolis, MN, USA
    • ,
  • Willem Ombelet

      Affiliations

    • Genk Institute for Fertility Technology (GIFT), Ziekenhuis Oost Limburg, Genk, Belgium

Received 24 March 2009 ,Revised 14 April 2009 ,Accepted 21 July 2009.

References 

  1. Aitken R. Sperm function test and fertility. International Journal of Fertility. 2006;29:69–75
  2. Aitken R, Buckingham D, West K, et al. Differential contribution of leucocytes and spermatozoa to the generation of reactive oxygen species in the ejaculates of oligozoospermic patients and fertile donors. Journal of Reproduction and Fertility. 1992;94:451–462
  3. Aoki V, Emery B, Liu L, et al. Protamine levels vary between individual sperm cells of infertile human males and correlate with viability and DNA integrity. Journal of Andrology. 2006;27:890–898
  4. Bakos H, Thompson J, Feil D, et al. Sperm DNA damage is associated with assisted reproductive technology pregnancy. International Journal of Andrology. 2008;31:518–526
  5. Benchaib M, Braun V, Lornage J, et al. Sperm DNA fragmentation decreases the pregnancy rate in an assisted reproductive technique. Human Reproduction. 2003;18:1023–1028
  6. Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society. Series B (Methodological). 1995;57:289–300
  7. Bianchi P, Manicardi G, Urner F, et al. Chromatin packaging and morphology in ejaculated human spermatozoa: evidence of hidden anomalies in normal spermatozoa. Molecular Human Reproduction. 1996;2:139–144
  8. Bianchi P, Manicardi G, Bizzaro D, et al. Effect of deoxyribonucleic acid protamination on fluorochrome staining and in situ nick-translation of murine and human mature spermatozoa. Biology of Reproduction. 1993;49:1083–1088
  9. Bungum M, Spano M, Humaidan P, et al. Sperm chromatin structure assay parameters measured after density gradient centrifugation are not predictive for the outcome of ART. Human Reproduction. 2008;23:4–10
  10. Bungum M, Humaidan P, Axmon A, et al. Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome. Human Reproduction. 2007;22:174–179
  11. Bungum M, Humaidan P, Spano M, et al. The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI. Human Reproduction. 2004;19:1401–1408
  12. Carrell D, Liu L. Altered protamine 2 expression is uncommon in donors of known fertility, but common among men with poor fertilizing capacity, and may reflect other abnormalities of spermiogenesis. Journal of Andrology. 2001;22:604–610
  13. Carrell D, Emery B, Hammoud S. Altered protamine expression and diminished spermatogenesis: what is the link?. Human Reproduction Update. 2007;13:313–327
  14. Cayli S, Sakkas D, Vigue L, et al. Cellular maturity and apoptosis in human sperm: creatine kinase, caspase-3 and Bcl-xl levels in mature and diminished maturity sperm. Molecular Human Reproduction. 2004;10:365–372
  15. Cayli S, Jakab A, Ovari L, et al. Biochemical markers of sperm function: male fertility and sperm selection for ICSI. Reproductive BioMedicine Online. 2003;7:462–468
  16. Cendeho A. Evaluation of the subfertile male. In:  Oehninger S,  Kruger T editor. Male Infertility: Diagnosis and Treatment. London: Informa; 2007;p. 117–140
  17. Check J, Graziano V, Cohen R, et al. Effect of an abnormal sperm chromatin structural assay (SCSA) on pregnancy outcome following (IVF) with ICSI in previous IVF failures. Archives of Andrology. 2005;51:121–124
  18. Collins J, Barnhart K, Schlegel P. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization?. Fertility and Sterility. 2008;89:823–831
  19. Darzynkiewicz Z, Traganos F, Sharpless T, et al. Thermal denaturation of DNA in situ as studied by acridine orange staining and automated cytofluorometry. Experimental Cell Research. 1975;90:411–428
  20. de Liu Y, Liu M, Garrett C, et al. Comparison of the frequency of defective sperm–zona pellucida (ZP) binding and the ZP-induced acrosome reaction between subfertile men with normal and abnormal semen. Human Reproduction. 2007;22:1878–1884
  21. De Yebra L, Ballesca J, Vanrell J, et al. Complete selective absence of protamine P2 in humans. Journal of Biochemical Chemistry. 1993;268:10553–10557
  22. Duran E, Morshedi M, Taylor S, et al. Sperm DNA quality predicts intrauterine insemination outcome: a prospective cohort study. Human Reproduction. 2002;17:3122–3128
  23. Erenpreiss J, Spano M, Erenpreisa J, et al. Sperm chromatin structure and male fertility: biological and clinical aspects. Asian Journal of Andrology. 2006;8:11–29
  24. Esterhuizen A, Franken D, Lourens J, et al. Chromatin packaging as an indicator of human sperm dysfunction. Journal of Assisted Reproduction and Genetics. 2000;17:508–514
  25. Esterhuizen A, Franken D, Lourens J, et al. Sperm chromatin packaging as an indicator of in-vitro fertilization rates. Human Reproduction. 2000;15:657–661
  26. Evenson D, Jost L. Sperm chromatin structure assay is useful for fertility assessment. Methods in Cell Science. 2000;22:169–189
  27. Evenson D, Jost L. Sperm chromatin structure assay: DNA denaturability. Methods in Cell Biology. 1994;42:159–176
  28. Evenson D, Wixon R. Meta-analysis of sperm DNA fragmentation using the sperm chromatin structure assay. Reproductive BioMedicine Online. 2006;12:466–472
  29. Evenson D, Larson K, Jost L. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. Journal of Andrology. 2002;23:25–43
  30. Evenson D, Jost L, Marshall D, et al. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Human Reproduction. 1999;14:1039–1149
  31. Franken D, Kruger T 2007 Sperm morphology training and quality control programs are essential for clinically relevant results Male Infertility: Diagnosis and Treatment. London: Informa; 2007. p. 181–186.
  32. Franken D, Franken C, de la Guerre H, et al. Normal sperm morphology and chromatin packaging: comparison between aniline blue and chromomycin A3 staining. Andrologia. 1999;31:361–366
  33. Gadella B, Visconti P 2006 Regulation of capacitation. In: De Jonge C, Barratt C, editors The Sperm Cell: Production, Maturation Fertilisation, Regeneration. Cambridge: Cambridge University Press; 2006. p. 134–169.
  34. Gandini L, Lombardo F, Paoli D, et al. Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage. Human Reproduction. 2004;19:1409–1417
  35. Gandini L, Lombardo F, Paoli D. Study of apoptotic DNA fragmentation in human spermatozoa. Human Reproduction. 2000;15:830–839
  36. Gázquez C, Oriola J, de Mateo S, et al. A common protamine 1 promoter polymorphism (−190 C→A) correlates with abnormal sperm morphology and increased protamine P1/P2 ratio in infertile patients. Journal of Andrology. 2008;25:540–548
  37. Ghosh I, Chattopadhaya R, Kumar V, et al. Hyaluronan binding protein-1: a modulator of sperm–oocyte interaction. Society of Reproduction and Fertility. 2007;63:539–543
  38. Giwercman A, Richthoff J, Hjollund H, et al. Correlation between sperm motility and sperm chromatin structure assay parameters. Fertility and Sterility. 2003;80:1404–1412
  39. Hammadeh M, Kühnen A, Amer A, et al. 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. International Journal of Andrology. 2001;24:360–368
  40. Hammadeh M, Stieber M, Haidl G, et al. Association between sperm cell chromatin condensation, morphology based on strict criteria, and fertilization, cleavage and pregnancy rates in an IVF program. Andrologia. 1998;30:29–35
  41. Henkel R 2007a Detection of DNA damage in sperm. In: Oehninger S, Kruger T, editors Male Infertility: Diagnosis and Treatment. London: Informa; 2007. p. 225–238.
  42. Henkel R 2007b DNA fragmentation and its influence on fertilisation and pregnancy outcome. In: Oehninger S, Kruger T, editors Male Infertility: Diagnosis and Treatment. London: Informa; 2007. p. 277–290.
  43. Hunnicutt G, Primakoff P, Myles D. Sperm surface protein PH-20 is bifunctional: one activity is a hyaluronidase and a second, distinct activity is required in secondary sperm–zona binding. Biology of Reproduction. 1996;55:80–86
  44. Huszar G, Ozenci C, Cayli S, et al. Hyaluronic acid binding by human sperm indicates cellular maturity, viability and unreacted acrosomal status. Fertility and Sterility. 2003;79(Suppl. 3):1616–1624
  45. Iranpour F, Nasr-Esfahani M, Valojerdi M, et al. Chromomycin A3 staining as a useful tool for evaluation of male infertility. Journal of Assisted Reproduction and Genetics. 2000;17:60–66
  46. Kornovski B, McCoshen J, Krdentser J, et al. The regulation of sperm motility by a novel hyaluronan receptor. Fertility and Sterility. 1994;61:935–940
  47. Kruger T, Menkveld R, Stander F, et al. Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertility and Sterility. 1986;46:1118–1123
  48. Larson K, DeJonge C, Barnes A, et al. Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Human Reproduction. 2000;15:1717–1722
  49. Lewis S. Is sperm evaluation useful in predicting human fertility?. Reproduction. 2007;134:1–11
  50. Lewis S, Agbaje S, Alvarez J. Sperm DNA tests as useful adjuncts to semen analysis. Systems Biology in Reproductive Medicine. 2008;54:111–125
  51. Lin M, Kuo-Kuang L, Li S, et al. Sperm chromatin structure assay parameters are not related to fertilisation rates, embryo quality, and pregnancy rates in in vitro fertilisation and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertility and Sterility. 2008;90:352–359
  52. Lolis D, Georgiou I, Syrrou M, et al. Chromomycin A3-staining as an indicator of protamine deficiency and fertilisation. International Journal of Andrology. 1996;19:23–27
  53. Manicardi G, Bianchi P, Pantano S, et al. Presence of endogenous nicks in DNA of ejaculated human spermatozoa and its relationship to chromomycin A3 accessibility. Biology of Reproduction. 1995;52:864–867
  54. Menkveld R 2007 The basic semen analysis. In: Oehninger S, Kruger T, editors Male Infertility: Diagnosis and Treatment. London: Informa; 2007. p. 141–170.
  55. Menkveld R, Oettle E, Kruger T et al. editors Atlas of Human Sperm Morphology, Baltimore: Williams & Wilkins; 1991.
  56. Morrell J, Moffatt O, Sakkas D, et al. Reduced senescence and retained nuclear DNA integrity in human spermatozoa prepared by density gradient centrifugation. Journal of Assisted Reproduction and Genetics. 2004;21:217–222
  57. Muriel L, Meseguer M, Fernandez J, et al. Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospective study. Human Reproduction. 2006;21:738–744
  58. Nanassy L, Carrell D. Paternal effects on early embryogenesis. Journal of Experimental and Clinical Assisted Reproduction. 2008;5:1–9
  59. Nasr-Esfahani M, Razavi S, Travalaee , et al. Failed fertilization after ICSI and spermiogenic defects. Fertility and Sterility. 2008;89:892–898
  60. Nasr-Esfahani M, Razavi S, Mardani M, et al. Effects of failed oocyte activation and sperm protamine deficiency on fertilization post-ICSI. Reproductive BioMedicine Online. 2007;14:422–429
  61. Nasr-Esfahani M, Salehi M, Razavi S, et al. Effect of sperm DNA damage and sperm protamine deficiency on fertilization and embryo development post-ICSI. Reproductive BioMedicine Online. 2005;11:198–205
  62. Nasr-Esfahani M, Salehi M, Razavi S, et al. Effect of protamine-2 deficiency on ICSI outcome. Reproductive BioMedicine Online. 2004;9:652–658
  63. Nijs M, Ombelet W 2004 Sperm analysis and reproduction update. In: Gregory L, Vanblerkom J, editors Essential IVF: Basic Research and Clinical Applications. Publishers, Boston: Kluwer; 2004. p. 151–178.
  64. Nijs M, Ombelet W. Cryopreservation of human sperm. Human Fertility. 2001;4:158–163
  65. Nijs M, Ombelet W. Intracytoplasmic sperm injection in assisted reproductive technology: an evaluation. Human Fertility. 2000;3:221–225
  66. Nijs M, Creemers E, Cox A, et al. Influence of freeze–thawing on hyaluronic acid binding of human spermatozoa. Reproductive BioMedicine Online. 2009;19:202–206
  67. Nijs M, Vanderzwalmen P, Segal-Bertin G, et al. A monozygotic twin pregnancy after zona rubbing on a frozen–thawed blastocyst. Human Reproduction. 1993;8:127–129
  68. Nijs M, Geerts l, Van Roosendaal E, et al. Prevention of multiple pregnancies in an in-vitro fertilization programme. Fertility and Sterility. 1993;59:1245–1250
  69. Nixon B, Aitken R, McLaughlin E. New insights into the molecular mechanisms of sperm-egg interaction. Cellular and Molecular Life Sciences. 2007;64:1805–1823
  70. Oehninger S. Biochemical and functional characterization of the zona pellucida. Reproductive BioMedicine Online. 2003;7:641–648
  71. Oehninger S, Arslan M, Franken D 2007 Sperm-zona pellucida binding assays. In: Oehninger S, Kruger T editors Male Infertility: Diagnosis and Treatment. London: Springer; 2007. p. 209–223.
  72. Oehninger S, Franken D, Sayed E, et al. Sperm function assays and their predictive value for fertilisation outcome in IVF therapy: a meta-analysis. Human Reproduction Update. 2000;6:160–168
  73. Oliva R. Protamines and male infertility. Human Reproduction Update. 2006;12:417–435
  74. Ombelet W, Deblare K, Bosmans E, et al. Semen quality and intrauterine insemination. Reproductive BioMedicine Online. 2003;7:485–492
  75. Ombelet W, Vandeput H, Van de Putte G, et al. Intrauterine insemination after ovarian stimulation with clomiphene citrate: predictive potential of inseminating motile count and sperm morphology. Human Reproduction. 1997;12:1458–1463
  76. Ombelet W, Menkveld R, Kruger KT, et al. Sperm morphology assessment: historical review in relation to fertility. Human Reproduction Update. 1995;1:543–557
  77. Prinosilova P, Kruger T, Sati L, et al. Selectivity of hyaluronic acid binding for spermatozoa with normal Tygerberg strict morphology. Reproductive BioMedicine Online. 2009;18:177–183
  78. Razavi S, Nasr-Esfahani M, Mardani M, et al. Effect of human sperm chromatin anomalies on fertilization outcome post-ICSI. Andrologia. 2003;35:238–243
  79. Sabeur K, Cherr G, Yudin A, et al. Hyaluronic acid enhances induction of the acrosome reaction of human sperm through interaction with the PH-20 protein. Zygote. 1998;6:103–111
  80. Sabeur K, Cherr G, Yudin A, et al. The PH-20 protein in human spermatozoa. Journal of Andrology. 1997;18:151–158
  81. Sakkas D, Manicardi G, Tomlinson M, et al. The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies. Human Reproduction. 2000;15:1112–1116
  82. Sakkas D, Urner F, Bianchi P, et al. Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection. Human Reproduction. 1996;11:837–843
  83. Saleh R, Agarwal A, Nada E, et al. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertility and Sterility. 2003;79(Suppl. 3):1597–1605
  84. Seli E, Gardner D, Schoolcraft W, et al. Extent of nuclear DNA damage in ejaculated spermatozoa impacts on blastocyst development after in vitro fertilisation. Fertility and Sterility. 2004;82:378–383
  85. Siebert T, van der Merwe F, Kruger T et al. 2007 How do we define male subfertility and what is the prevalence in the general population? In: Oehninger S, Kruger T, editors Male Infertility: Diagnosis and Treatment. London: Informa; 2007. p. 269–276.
  86. Spano M, Bonde J, Hjollund H, et al. Sperm chromatin damage impairs human fertility. The Danish First Pregnancy Planner Study Team. Fertility and Sterility. 2000;73:43–50
  87. Spano M, Cordelli E, Leter G, et al. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay. Molecular Human Reproduction. 1999;5:29–37
  88. Singleton S, Mudrak O, Morshedi M, et al. Characterisation of a human sperm cell subpopulation marked by the presence of the TSH2B histone. Reproduction, Fertility, and Development. 2007;19:392–399
  89. Tarozzi N, Bizzaro D, Flamigni C, et al. Clinical relevance of sperm DNA damage in reproduction. Reproductive BioMedicine Online. 2007;14:746–757
  90. Tavalaee M, Razavi S, Nasr-Esfahani M. Influence of sperm chromatin anomalies on assisted reproductive technology outcome. Fertility and Sterility. 2008;Epub ahead of print
  91. Tomlinson M, Moffatt O, Manicardi G, et al. Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: implications for assisted conception. Human Reproduction. 2001;16:2160–2165
  92. Torregrosa D, Domínguez-Fandos D, Camejo M, et al. Protamine 2 precursors, protamine 1/protamine 2 ratio, DNA integrity and other sperm parameters in infertile patients. Human Reproduction. 2006;21:2084–2089
  93. Twigg J, Irvine D, Aitken R, et al. Oxidative damage to DNA in human spermatozoa does not preclude pronucleus formation at intracytoplasmic sperm injection. Human Reproduction. 1998;13:1864–1871
  94. Virro M, Larson-Cook K, Evenson D. Sperm chromatin structure assay (SCSA) parameters are related to fertilisation, blastocyst development, and ongoing pregnancy in in vitro fertilisation and intracytoplasmic sperm injection cycles. Fertility and Sterility. 2004;81:1289–1295
  95. Vjugina U, Evans J. New insights into the molecular basis of mammalian sperm–egg membrane interactions. Frontiers in Bioscience. 2008;13:462–476
  96. World Health Organization 1999 WHO Laboratory Manual for the Examination of Human Semen and Sperm–Cervical Mucus Interaction. Cambridge University Press, Cambridge.
  97. Ye H, Huang G, Gao Y, et al. Relationship between human sperm-hyaluronan binding assay and fertilisation rate in conventional in vitro fertilisation. Human Reproduction. 2006;21:1545–1550
  98. Zini A, Sigman M. Are tests of sperm DNA damage clinically useful? Pros and Cons. Journal of Andrology. 2009;30:219–229
  99. Zini A, Philips S, Courchesne A, et al. Sperm head morphology is related to high deoxyribonucleic stainability assessed by sperm chromatin structure assay. Fertility and Sterility. 2008;Epub ahead of print
  100. Zini A, Zhang X, San Gabriel M. Sperm nuclear histone H2B: correlation with sperm DNA denaturation and DNA stainability. Asian Journal of Andrology. 2008;10:865–871
  101. Zini A, Boman J, Belzile E. Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Human Reproduction. 2008;23:2663–2668
  102. Zini A, Bielecki R, Phang D. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertility and Sterility. 2001;75:674–677
  103. Zhang X, SanGabriel M, Zini A. Sperm nuclear histone to protamine ratio in fertile and infertile men: evidence of heterogeneous subpopulations of spermatozoa in the ejaculate. Journal of Andrology. 2006;27:414–420

 Martine Nijs obtained her PhD in Medical Sciences at the Free University of Brussels in Belgium. She is ESHRE accredited senior clinical embryologist. At present, she is the director of the IVF laboratory at the Genk Institute for Fertility Technology in Genk, Belgium. Her main interests are sperm quality, sperm freezing and human embryology. Dr Martine Nijs

☆ Partially presented as an oral communication (0–165) at the 21st Meeting of ESHRE, Copenhagen, Denmark, 19–22, June 2005.Partially presented as a poster at the 31st Annual Meeting of the American Society of Andrology, Chicago, USA, April 8–11, 2006.Partially presented as a poster at the Satellite Sperm DNA workshop in Rome, 11–13 March, 2009.

PII: S1472-6483(09)00064-9

doi: 10.1016/j.rbmo.2009.07.002

Reproductive BioMedicine Online
Volume 19, Issue 5 , Pages 671-684 , November 2009

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