DOI QR코드

DOI QR Code

The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis

  • Kim, Hee-Sun (IVF Laboratory, Department of Obstetrics and Gynecology, Seoul National University Hospital) ;
  • Kang, Moon Joo (IVF Laboratory, Department of Obstetrics and Gynecology, Seoul National University Hospital) ;
  • Kim, Sung Ah (IVF Laboratory, Department of Obstetrics and Gynecology, Seoul National University Hospital) ;
  • Oh, Sun Kyung (IVF Laboratory, Department of Obstetrics and Gynecology, Seoul National University Hospital) ;
  • Kim, Hoon (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Ku, Seung-Yup (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Kim, Seok Hyun (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Moon, Shin Yong (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Choi, Young Min (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University)
  • Received : 2012.12.13
  • Accepted : 2013.01.16
  • Published : 2013.03.31

Abstract

Objective: The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation. Methods: Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests. Results: Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029). Conclusion: The tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.

Keywords

References

  1. World Health Organization. WHO laboratory manual for the examination and processing of human semen [Internet]. 5th ed. Geneva: World Health Organization; c2013 [cited 2013 Mar 4]. Available from: http://www.who.int/reproductivehealth/publications/ infertility/9789241547789/en/index.html.
  2. Guzick DS, Overstreet JW, Factor-Litvak P, Brazil CK, Nakajima ST, Coutifaris C, et al. Sperm morphology, motility, and concentration in fertile and infertile men. N Engl J Med 2001;345:1388-93. https://doi.org/10.1056/NEJMoa003005
  3. Evenson DP, Jost LK, Marshall D, Zinaman MJ, Clegg E, Purvis K, et al. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 1999;14:1039-49. https://doi.org/10.1093/humrep/14.4.1039
  4. Carrell DT, Liu L, Peterson CM, Jones KP, Hatasaka HH, Erickson L, et al. Sperm DNA fragmentation is increased in couples with unexplained recurrent pregnancy loss. Arch Androl 2003;49:49-55. https://doi.org/10.1080/01485010290099390
  5. Virro MR, Larson-Cook KL, Evenson DP. Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril 2004; 81:1289-95. https://doi.org/10.1016/j.fertnstert.2003.09.063
  6. Talebi AR, Vahidi S, Aflatoonian A, Ghasemi N, Ghasemzadeh J, Firoozabadi RD, et al. Cytochemical evaluation of sperm chromatin and DNA integrity in couples with unexplained recurrent spontaneous abortions. Andrologia 2012;44 Suppl 1:462-70. https://doi.org/10.1111/j.1439-0272.2011.01206.x
  7. Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization? Fertil Steril 2008;89:823-31. https://doi.org/10.1016/j.fertnstert.2007.04.055
  8. Bellver J, Meseguer M, Muriel L, Garcia-Herrero S, Barreto MA, Garda AL, et al. Y chromosome microdeletions, sperm DNA fragmentation and sperm oxidative stress as causes of recurrent spontaneous abortion of unknown etiology. Hum Reprod 2010;25: 1713-21. https://doi.org/10.1093/humrep/deq098
  9. Marcon L, Boissonneault G. Transient DNA strand breaks during mouse and human spermiogenesis new insights in stage specificity and link to chromatin remodeling. Biol Reprod 2004;70: 910-8. https://doi.org/10.1095/biolreprod.103.022541
  10. Agarwal A, Erenpreiss J, Sharma R. Sperm chromatin assessment. In: Gardner DK, Weissman A, Howles CM, Shoham Z, editors. Textbook of assisted reproductive technologies. 3rd ed. London: Informa Healthcare; 2009. p. 67-84.
  11. Dadoune JP, Mayaux MJ, Guihard-Moscato ML. Correlation between defects in chromatin condensation of human spermatozoa stained by aniline blue and semen characteristics. Andrologia 1988;20:211-7.
  12. Braun RE. Packaging paternal chromosomes with protamine. Nat Genet 2001;28:10-2.
  13. Erenpreiss J, Bars J, Lipatnikova V, Erenpreisa J, Zalkalns J. Comparative study of cytochemical tests for sperm chromatin integrity. J Androl 2001;22:45-53.
  14. Erenpreiss J, Jepson K, Giwercman A, Tsarev I, Erenpreisa J, Spano M. Toluidine blue cytometry test for sperm DNA conformation: comparison with the flow cytometric sperm chromatin structure and TUNEL assays. Hum Reprod 2004;19:2277-82. https://doi.org/10.1093/humrep/deh417
  15. Wong A, Chuan SS, Patton WC, Jacobson JD, Corselli J, Chan PJ. Addition of eosin to the aniline blue assay to enhance detection of immature sperm histones. Fertil Steril 2008;90:1999-2002. https://doi.org/10.1016/j.fertnstert.2007.09.026
  16. Park YS, Kim MK, Lee SH, Cho JW, Song IO, Seo JT. Efficacy of testicular sperm chromatin condensation assay using aniline blueeosin staining in the IVF-ET cycle. Clin Exp Reprod Med 2011;38: 142-7. https://doi.org/10.5653/cerm.2011.38.3.142
  17. de la Rochebrochard E, Thonneau P. Paternal age and maternal age are risk factors for miscarriage; results of a multicentre European study. Hum Reprod 2002;17:1649-56. https://doi.org/10.1093/humrep/17.6.1649
  18. Barratt CL, Aitken RJ, Bjorndahl L, Carrell DT, de Boer P, Kvist U, et al. Sperm DNA: organization, protection and vulnerability: from basic science to clinical applications--a position report. Hum Reprod 2010;25:824-38. https://doi.org/10.1093/humrep/dep465
  19. The Practice Committee of the American Society for Reproductive Medicine. The clinical utility of sperm DNA integrity testing. Fertil Steril 2008; 90:S178-80.
  20. The Practice Committee of the American Society for Reproductive Medicine. Diagnostic evaluation of the infertile male: a committee opinion. Fertil Steril 2012;98:294-301. https://doi.org/10.1016/j.fertnstert.2012.05.033
  21. Hammadeh ME, Zeginiadov T, Rosenbaum P, Georg T, Schmidt W, Strehler E. Predictive value of sperm chromatin condensation (aniline blue staining) in the assessment of male fertility. Arch Androl 2001;46:99-104.
  22. Nijs M, De Jonge C, Cox A, Janssen M, Bosmans E, Ombelet W. Correlation between male age, WHO sperm parameters, DNA fragmentation, chromatin packaging and outcome in assisted reproduction technology. Andrologia 2011;43:174-9. https://doi.org/10.1111/j.1439-0272.2010.01040.x
  23. Beshay VE, Bukulmez O. Sperm DNA damage: how relevant is it clinically? Curr Opin Obstet Gynecol 2012;24:172-9. https://doi.org/10.1097/GCO.0b013e32835211b5

Cited by

  1. Assessment of Chromatin Maturity in Human Spermatozoa: Useful Aniline Blue Assay for Routine Diagnosis of Male Infertility vol.2013, pp.None, 2013, https://doi.org/10.1155/2013/578631
  2. The Usefulness of Selected Physicochemical Indices, Cell Membrane Integrity and Sperm Chromatin Structure in Assessments of Boar Semen Sensitivity vol.28, pp.12, 2013, https://doi.org/10.5713/ajas.15.0095
  3. Investigation of the association between the outcomes of sperm chromatin condensation and decondensation tests, and assisted reproduction techniques vol.47, pp.4, 2015, https://doi.org/10.1111/and.12286
  4. A human morphologically normal spermatozoon may have noncondensed chromatin vol.47, pp.8, 2015, https://doi.org/10.1111/and.12341
  5. Effect of metformin therapy and dietary supplements on semen parameters in hyperinsulinaemic males vol.47, pp.9, 2013, https://doi.org/10.1111/and.12366
  6. Effect of chronic pesticide exposure on murine cornea: a histopathological, cytological and flow cytometric approach to study ocular damage by xenobiotics vol.32, pp.1, 2016, https://doi.org/10.1007/s10565-016-9314-4
  7. Optimization of microelectrophoresis to select highly negatively charged sperm vol.33, pp.6, 2013, https://doi.org/10.1007/s10815-016-0700-x
  8. Deregulation of vital mitotic kinase-phosphatase signaling in hematopoietic stem/progenitor compartment leads to cellular catastrophe in experimental aplastic anemia vol.422, pp.1, 2013, https://doi.org/10.1007/s11010-016-2811-1
  9. Mammalian sperm nuclear organization: resiliencies and vulnerabilities vol.26, pp.1, 2013, https://doi.org/10.1186/s12610-016-0044-5
  10. Profile of MicroRNA Expression in Endometrial Cell during In Vitro Culture According to Progesterone Concentration vol.14, pp.5, 2017, https://doi.org/10.1007/s13770-017-0080-z
  11. Eight tests for sperm DNA fragmentation and their roles in the clinic vol.6, pp.4, 2013, https://doi.org/10.21037/tau.2017.03.78
  12. Impact of hormonal changes on the semen quality and assisted reproductive outcomes in infertile men vol.15, pp.3, 2013, https://doi.org/10.1016/j.jab.2017.04.003
  13. Red light improves spermatozoa motility and does not induce oxidative DNA damage vol.7, pp.None, 2013, https://doi.org/10.1038/srep46480
  14. A comparative evaluation of migration sedimentation method for sperm preparation vol.64, pp.2, 2013, https://doi.org/10.1080/19396368.2017.1402100
  15. Growth and male reproduction improvement of non-thermal dielectric barrier discharge plasma treatment on chickens vol.51, pp.20, 2018, https://doi.org/10.1088/1361-6463/aabd9a
  16. Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels vol.8, pp.None, 2018, https://doi.org/10.1038/s41598-018-26049-5
  17. Sperm selection for assisted reproduction by prior hyaluronan binding: the HABSelect RCT vol.6, pp.1, 2013, https://doi.org/10.3310/eme06010
  18. Nuclear sperm quality in total polymorphic teratozoospermia and its impact on intracytoplasmic sperm injection outcome vol.51, pp.5, 2013, https://doi.org/10.1111/and.13252
  19. Assessment of different sperm functional tests in golden‐headed lion tamarins (Leontopithecus chrysomelas) vol.81, pp.8, 2019, https://doi.org/10.1002/ajp.23034
  20. Decreased bull fertility: age-related changes in sperm motility and DNA fragmentation vol.151, pp.None, 2020, https://doi.org/10.1051/e3sconf/202015101010
  21. Manual Collection and Semen Characterization in a West Indian Manatee ( Trichechus manatus ) vol.7, pp.None, 2013, https://doi.org/10.3389/fvets.2020.569993
  22. Semiquantitative promoter methylation of MLH1 and MSH2 genes and their impact on sperm DNA fragmentation and chromatin condensation in infertile men vol.53, pp.1, 2013, https://doi.org/10.1111/and.13827
  23. Comparative analysis of tests used to assess sperm chromatin integrity and DNA fragmentation vol.53, pp.2, 2021, https://doi.org/10.1111/and.13718
  24. Dolichandrone serrulata flower extract ameliorates male reproductive damages in type 2 diabetic rats vol.53, pp.2, 2021, https://doi.org/10.1111/and.13911
  25. Idiopathic Infertility as a Feature of Genome Instability vol.11, pp.7, 2013, https://doi.org/10.3390/life11070628
  26. Seminal Plasma Protein N-Glycan Peaks Are Potential Predictors of Semen Pathology and Sperm Chromatin Maturity in Men vol.11, pp.9, 2013, https://doi.org/10.3390/life11090989
  27. Effect of Sperm Dna Fragmentation Index on Clinical Outcomes of Intra-Uterine Insemination Patients vol.14, pp.3, 2013, https://doi.org/10.13005/bpj/2230
  28. Correlation between Sperm Dna Fragmentation and Conventional Semen Parameters among Different Age Groups vol.14, pp.3, 2013, https://doi.org/10.13005/bpj/2235