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Chapter 21 - DNA Damage: COMET Assay

Published online by Cambridge University Press:  05 April 2021

Ashok Agarwal
The Cleveland Clinic Foundation, Cleveland, OH
Ralf Henkel
University of the Western Cape, South Africa
Ahmad Majzoub
Hamad Medical Corporation, Doha
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Sperm are highly specialized cells, evolved to function as vehicles for the transport of the paternal genome to the oocyte. The sperm cell is characterized by a distinct head, mid-piece and tail, structured for a streamlined function. The sperm head consists of the haploid paternal genome (23 chromosomes), packed in a specific tight manner with the help of specialized proteins called protamines. The mid-piece consists of the centrosome and mitochondria, organelles that provide energy for sperm propulsion from the tail. The unique sperm structure, complimented with its motility, helps the sperm to swim through the male and female reproductive tract and penetrate the egg. Therefore, the primary function of the sperm is to successfully deliver the paternal genome to the oocyte.

Publisher: Cambridge University Press
Print publication year: 2021

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Aitken, RJ, de Iuliis, GN. On the possible origins of DNA damage in human spermatozoa. Mol Hum Reprod 2010; 16: 313.Google Scholar
Simon, L, Lutton, D, McManus, J, Lewis, SEM. Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and IVF success. Fertil Steril 2011a; 95: 652–7.CrossRefGoogle Scholar
Panner Selvam, MK, Agarwal, A. A systematic review on sperm DNA fragmentation in male factor infertility: laboratory assessment. Arab J Urol 2018; 16(1): 6576.Google Scholar
Singh, NR, Stephens, RE. X-ray-induced DNA double-strand breaks in human sperm. Mutagenesis 1998; 13(1): 75–9.Google Scholar
Simon, L, Aston, KI, Emery, BR, Hotaling, J, Carrell, DT. Sperm DNA damage output parameters measured by the alkaline Comet assay and their importance. Andrologia 2017a; 49: e12608.CrossRefGoogle ScholarPubMed
Shamsi, MB, Kumar, R, Dada, R. Evaluation of nuclear DNA damage in human spermatozoa in men opting for assisted reproduction. Indian J Med Res 2008; 127: 115–23.Google Scholar
Shamsi, MB, Imam, SN, Dada, R. Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in management of infertility. J Assist Reprod Genet 2011; 28: 1073–85.Google Scholar
Olive, PL, Wlodek, D, Durand, RE, Banáth, JP. Factors influencing DNA migration from individual cells subjected to gel electrophoresis. Exp Cell Res 1992; 198: 259–67.CrossRefGoogle ScholarPubMed
Hughes, CM, Lewis, SEM, McKelvey-Martin, V, Thompson, W. Reproducibility of human sperm DNA measurements using the alkaline single cell gel electrophoresis assay. Mutat Res 1997; 374: 261–8.Google Scholar
Morris, ID, Iiott, S, Dixon, L, Brison, DR. 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 2002; 17(4): 990–8.CrossRefGoogle ScholarPubMed
Piperakis, SM. Comet assay: a brief history. Cell Biol Toxicol 2009; 25: 13.CrossRefGoogle ScholarPubMed
Ribas-Maynou, J, Fernandez-Encinas, A, Garcıa-Peiro, A, Prada, E, Abad, C, Amengual, MJ, Navarro, J, Benet, J. Human semen cryopreservation: a sperm DNA fragmentation study with alkaline and neutral Comet assay. Andrology 2014; 2: 83–7.CrossRefGoogle ScholarPubMed
Simon, L, Murphy, K, Shamsi, MB, Liu, L, Emery, B, Aston, KI, Hotaling, J, Carrell, DT. Paternal influence of sperm DNA integrity on early embryonic development. Hum Reprod 2014b; 29(11): 2402–12.Google Scholar
Simon, L, Brunborg, G, Stevenson, M, Lutton, D, McManus, J, Lewis, SEM. Clinical significance of sperm DNA damage in assisted reproductive outcome. Hum Reprod 2010; 25(7): 1594–608.CrossRefGoogle Scholar
Simon, L, Liu, L, Murphy, K, Ge, S, Hotaling, J, Aston, KI, Emery, B, Carrell, DT. Comparative analysis of three sperm DNA damage assays and sperm nuclear protein content in couples undergoing assisted reproduction treatment. Hum Reprod 2014a; 29: 904–17.Google Scholar
Simon, L, Carrell, DT. ( 2011d) Sperm DNA damage measured by Comet assay. In Carrell, DT and Aston, KI, Spermatogenesis and Spermiogenesis: Methods and Protocols. New Jersey: Humana Press, pp. 137–46.Google Scholar
Simon, L, Castillo, J, Oliva, R, Lewis, S. The relationship between human sperm protamines, DNA damage and assisted reproductive outcomes. Reprod Biomed Online 2011b; 23: 724–34.CrossRefGoogle Scholar
Dorostghoal, M, Kazeminejad, SR, Shahbazian, N, Pourmehdi, M, Jabbari, A. Oxidative stress status and sperm DNA fragmentation in fertile and infertile men. Andrologia 2017; 49(10): e12762.Google Scholar
Santi, D, Spaggiari, G, Simoni, M. Sperm DNA fragmentation index as a promising predictive tool for male infertility diagnosis and treatment management – meta-analyses. Reprod BioMed Online 2018; 37(3): 315–25.CrossRefGoogle ScholarPubMed
Sakkas, D. Novel technologies for selecting the best sperm for in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril 2013; 99: 1023–9.Google Scholar
Lewis, SEM, O’Connell, M, Stevenson, M, Thompson-Cree, L, McClure, N. An algorithm to predict pregnancy in assisted reproduction. Hum Reprod 2004; 19: 1385–94.Google Scholar
Nasr-Esfahani, MH, Salehi, M, Razavi, S, Anjlmshoa, M, Rozahani, S, Moulavi, F, Mardani, M. Effect of sperm DNA damage and sperm protamine deficiency on fertilization and embryo development post-ICSI. RBM Online 2005; 11: 198–205.Google ScholarPubMed
Tomsu, M, Sharma, V, Miller, D. Embryo quality and IVF treatment outcome may correlate with different sperm comet parameters. Hum Reprod 2002; 17: 1856–62.CrossRefGoogle Scholar
Simon, L, Zini, A, Ciampi, A, Dyachenko, A, Carrell, DT. A systematic review and meta-analysis to determine the effect of sperm DNA damage measured by different assays on IVF and ICSI outcomes. Asian J Androl 2017b; 19: 8090.Google Scholar
Bazrgar, M, Gourabi, H, Yazdi, PE, Vazirinasab, H, Fakhri, M, Hassani, F, Valojerdi, MR. DNA repair signalling pathway genes are overexpressed in poor-quality pre-implantation human embryos with complex aneuploidy. Eur J Obstet Gynecol Reprod Biol 2014; 175: 152–6.Google Scholar
Giwercman, A, Lindstedt, L, Larsson, M, Bungum, M, Spano, M, Levine, RJ, Rylander, L. Sperm chromatin structure assay as an independent predictor of fertility in vivo: a case-control study. Int J Androl 2010; 32: 221–7.Google Scholar
Simon, L, Lewis, SEM. Effects of progressive motility and sperm DNA damage on fertilization rates in vitro: is one better than the other? Syst Biol Reprod Med 2011c; 57(3): 133–8.Google Scholar
Evenson, DP, Jost, LK, Marshall, D, Zinaman, MJ, Clegg, E, Purvis, K, de Angelis, P, Claussen OP. Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 1999; 14: 1039–49.Google Scholar
Simon, L, Proutski, I, Stevenson, M, Jennings, D, McManus, J, Lutton, D, Lewis, SEM. Sperm DNA damage has negative association with live birth rates after IVF. Reprod Biomed Online 2013; 26: 6878.CrossRefGoogle ScholarPubMed
Simon, L, Carrell, DT, Zini, A. (2018) Sperm DNA tests are clinically useful. In Zini, A. and Agarwal, A., eds., A Clinician’s Guide to Sperm DNA and Chromatin Damage. New York: Springer International Publishing, pp. 431–67.Google Scholar
Oleszczuk, K, Augustinsson, L, Bayat, N, Giwercman, A, Bungum, M. Prevalence of high DNA fragmentation index in male partners of unexplained infertile couples. Andrology 2013; 1: 357–60.Google Scholar
Feijo, CM, Esteves, SC. Diagnostic accuracy of sperm chromatin dispersion test to evaluate sperm deoxyribonucleic acid damage in men with unexplained infertility. Fertil Steril 2014; 101: 5863.CrossRefGoogle ScholarPubMed

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