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Lack of an Association Between Sperm Head Abnormality and DNA Damage by Alkaline Comet Assay

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Sperm DNA damage affects sperm function, compromising reproductive outcome in both natural and assisted reproduction. The present study addresses the existing ambiguity between sperm morphology and DNA damage, by use of comet assay/single cell gel electrophoresis. Using comet assay as an end point, sperm DNA fragmentation in a total of 17 control and infertile subjects presenting to the infertility clinic of Kasturba Medical College has been investigated. Percent tail DNA, tail length and olive tail moment were selected as a measure to compare the extent of DNA damage between the groups. No significant difference was observed between the control group with morphologically normal sperm and experimental group with abnormal sperm morphology with respect to head DNA, tail DNA, Olive tail moment or tail length by alkaline comet assay. The comet assay failed to demonstrate any positive association between sperm morphology and DNA damage. In view of accumulating evidence on the importance of sperm DNA integrity in fertilization and embryogenesis, refined techniques allowing sperm selection with intact sperm DNA or with minimal DNA damage for clinical use should be sought.

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References

  1. Aitken RJ, De Iuliis GN (2007) Origins and consequences of DNA damage in male germ cells. Reprod Biomed Online 14:727–733

    Article  CAS  Google Scholar 

  2. Spano M, Bonde JP, Hjollund HI, Kolstad HA, Cordelli E, Leter G (2000) Sperm chromatin damage impairs human fertility. The Danish First Pregnancy Planner Study Team. Fertil Steril 73:43–50

    Article  CAS  Google Scholar 

  3. Morris I, Ilott S, Dixon L, Brison D (2002) The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (comet assay) and its relationship to fertilization and embryo development. Hum Reprod 17:990–998

    Article  CAS  Google Scholar 

  4. Benchaib M, Braun V, Lornage J, Hadj S, Salle B, Lejeune H et al (2003) Sperm DNA fragmentation decrease the pregnancy rate in an assisted reproductive technique. Hum Reprod 18:1023–1028

    Article  Google Scholar 

  5. Zini A, Boman JM, Belzile E, Ciampi A (2008) Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Hum Reprod 23:2663–2668

    Article  CAS  Google Scholar 

  6. Maher ER, Afnan M, Barratt CL (2003) Epigenetic risks related to assisted reproductive technologies: epigenetics, imprinting, ART and icebergs? Hum Reprod 18:2508–2511

    Article  Google Scholar 

  7. Avendaño C, Franchi A, Duran H, Oehninger S (2010) DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome. Fertil Steril 94:549–557

    Article  Google Scholar 

  8. Berkovitz A, Eltes F, Paul M, Adrian E, Benjamin B (2007) The chance of having a healthy normal child following intracytoplasmic morphologically selected sperm injection (IMSI) treatment is higher compared to conventional IVF-ICSI treatment. Fertil Steril 88:S20

    Article  Google Scholar 

  9. Antinori M, Licata E, Dani G, Cerusico F, Versaci C, d’Angelo D, Antinori S (2008) Intracytoplasmic morphologically selected sperm injection: a prospective randomized trial. Reprod Biomed Online 16:835–841

    Article  Google Scholar 

  10. Tasaka A, Doshida M, Sato Y, Kyoya T, Nakajo Y, Kyono K (2009) Outcome of IMSI (intracytoplasmic morpholocially selected sperm injection) in patients with repeated ICSI failures. Fertil Steril 92:S76

    Article  Google Scholar 

  11. Tesarik J (2005) Paternal effects on cell division in the human preimplantation embryo. Reprod Biomed Online 10:370–375

    Article  Google Scholar 

  12. Varshini J, Srinag BS, Kalthur G, Krishnamurthy H, Kumar P, Rao SB, Adiga SK (2012) Poor sperm quality and advancing age are associated with increased sperm DNA damage in infertile men. Andrologia 44:642–649

    Article  Google Scholar 

  13. World Health Organization (1999) Laboratory manual for the examination of human semen and sperm–cervical mucus interaction, 4th edn. Cambridge University Press, Cambridge

    Google Scholar 

  14. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  Google Scholar 

  15. Cary WH (1930) Sterility diagnosis: the study of sperm cell migration in female secretion and interpretation of findings. N Y State J Med 30:131–136

    Google Scholar 

  16. Joel CA (1971) Historical survey of research on spermatozoa from antiquity to the present. In: Joel CA (ed) Fertility disturbances in men and women. Karger, Basel, pp 3–47

    Google Scholar 

  17. Freund M (1966) Standards for the rating of human sperm morphology. A co-operative study. Int J Fertil 11:97–110

    Article  CAS  Google Scholar 

  18. Freund M (1968) Performance and interpretation of the semen analysis. In: Rolands M (ed) Management of the infertile couple. Charles C Thomas Publisher, Springfield, p 48

    Google Scholar 

  19. Van Zyl JA, Kotze TJvW, Menkveld R (1990) Predictive value of spermatozoa morphology in natural fertilization. In: Acosta AA, Swanson RJ, Ackerman SB, Kruger TF, van Zyl JA, Menkveld R (eds) Human spermatozoa in assisted reproduction. Williams and Wilkins, Baltimore, pp 319–324

    Google Scholar 

  20. Mahadevan MM, Trounson AO (1984) The influence of seminal characteristics on the success rate of human in vitro fertilization. Fertil Steril 42:400–405

    Article  CAS  Google Scholar 

  21. Taşdemir I, Taşdemir M, Tavukçuoglu S, Kahraman S, Biberoģlu K (1997) Effect of abnormal sperm head morphology on the outcome of intracytoplasmic sperm injection in humans. Hum Reprod 12:1214–1217

    Article  Google Scholar 

  22. Moomjy M, Sills ES, Rosenwaks Z, Palermo GD (1998) Implications of complete fertilization failure after intracytoplasmic sperm injection for subsequent fertilization and reproductive outcome. Hum Reprod 13:2212–2216

    Article  CAS  Google Scholar 

  23. O’Neil J, Lundquist L, Ritter J, Chantilis SJ, Carr BR, Byrd W (1998) Should strict morphology be used to indicate the need for intracytoplasmic sperm injection (ICSI)? Fertil Steril 70:S442

    Google Scholar 

  24. De Vos A, Van De Velde H, Joris H, Verheyen G, Devroey P, Van Steirteghem A (2003) Influence of individual sperm morphology on fertilization, embryo morphology, and pregnancy outcome of intracytoplasmic sperm injection. Fertil Steril 79:42–48

    Article  Google Scholar 

  25. Devillard F, Metzler-Guillemain C, Pelletier R, DeRobertis C, Bergues U, Hennebicq S, Guichaoua M, Sèle B, Rousseaux S (2002) Polyploidy in large-headed sperm: FISH study of three cases. Hum Reprod 17:1292–1298

    Article  CAS  Google Scholar 

  26. Barroso G, Oehninger SC (2007) Contribution of the male gamete to fertilization and embryogenesis. In: Oehiniger SC, Kruger TF (eds) Male infertility. Diagnosis and treatment. Informa Healthcare, London, pp 49–72

    Chapter  Google Scholar 

  27. Evenson D, Wixon R (2006) Meta-analysis of sperm DNA fragmentation using the sperm chromatin structure assay. Reprod Biomed Online 12:466–472

    Article  CAS  Google Scholar 

  28. Bungum M, Humaidan P, Axmon A, Spano M, Bungum L, Erenpreiss J et al (2007) Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome. Hum Reprod 22:174–179

    Article  CAS  Google Scholar 

  29. Ahmadi A, Ng SC (1999) Developmental capacity of damaged spermatozoa. Hum Reprod 14:2279–2285

    Article  CAS  Google Scholar 

  30. Elshal MF, El-Sayed IH, Elsaied MA, El-Masry SA, Kumosani TA (2009) Sperm head defects and disturbances in spermatozoal chromatin and DNA integrities in idiopathic infertile subjects: association with cigarette smoking. Clin Biochem 42:589–594

    Article  CAS  Google Scholar 

  31. Kahraman S, Akarsu C, Cengiz G, Dirican K, Sözen E, Can B, Güven C, Vanderzwalmen P (1999) Fertility of ejaculated and testicular megalohead spermatozoa with intracytoplasmic sperm injection. Hum Reprod 14:726–730

    Article  CAS  Google Scholar 

  32. Osawa Y, Sueoka K, Iwata S, Shinohara M, Kobayashi N, Kuji N, Yoshimura Y (1999) Assessment of the dominant abnormal form is useful for predicting the outcome of intracytoplasmic sperm injection in the case of severe teratozoospermia. J Assist Reprod Genet 16:436–442

    Article  CAS  Google Scholar 

  33. Garolla A, Fortini D, Menegazzo M, De Toni L, Nicoletti V, Moretti A, Selice R, Engl B, Foresta C (2008) High-power microscopy for selecting spermatozoa for ICSI by physiological status. Reprod Biomed Online 17:610–616

    Article  Google Scholar 

  34. Oliveira JB, Massaro FC, Baruffi RL, Mauri AL, Petersen CG, Silva LF, Vagnini LD, Franco JG Jr (2010) Correlation between semen analysis by motile sperm organelle morphology examination and sperm DNA damage. Fertil Steril 94:1937–1940

    Article  CAS  Google Scholar 

  35. Trivedi PP, Kushwaha S, Tripathi DN, Jena GB (2010) Evaluation of male germ cell toxicity in rats: correlation between sperm head morphology and sperm comet assay. Mutat Res 703:115–121

    Article  CAS  Google Scholar 

  36. Auger J, Eustache F, Andersen AG, Irvine DS, Jørgensen N, Skakkebaek NE, Suominen J, Toppari J, Vierula M, Jouannet P (2001) Sperm morphological defects related to environment, lifestyle and medical history of 1001 male partners of pregnant women from four European cities. Hum Rep 16:2710–2717

    Article  CAS  Google Scholar 

  37. Jakab A, Sakkas D, Delpiano E, Cayli S, Kovanci E, Ward D, Revelli A, Huszar G (2005) Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies. Fertil Steril 84:1665–1673

    Article  Google Scholar 

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Acknowledgement

Funding from Indian Council of Medical Research in the form of ad hoc scheme (5/10/7/2003-RHN) to SKA and Senior Research Fellowship to VJ (Nos. 3/1/2/5/2010-RHN, IRIS ID 2009-06740) is gratefully acknowledged. The authors thank Jayalaxmi Pai, Sandhya Patil and Keerthi G Patil, from the Manipal Assisted Reproduction Centre for the technical support.

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  1. Varshini Jayaraman

    Present address: Department of Animal Biology, School of Life Sciences, University of Hyderabad, Hyderabad, 500 046, India

Authors and Affiliations

  1. Division of Clinical Embryology, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Varshini Jayaraman, Dinesh Upadhya & Satish Kumar Adiga

  2. Department of Obstetrics and Gynecology, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Pratap Kumar Narayan

  3. Department of Anatomy, Kasturba Medical College, Manipal University, Manipal, 576 104, India

    Dinesh Upadhya

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  1. Varshini Jayaraman
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  4. Satish Kumar Adiga
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Correspondence to Varshini Jayaraman.

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Jayaraman, V., Upadhya, D., Narayan, P.K. et al. Lack of an Association Between Sperm Head Abnormality and DNA Damage by Alkaline Comet Assay. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 1345–1350 (2018). https://doi.org/10.1007/s40011-017-0863-y

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  • DOI: https://doi.org/10.1007/s40011-017-0863-y

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