DNA Damage: Fluorescent In-Situ Hybridization

doi:10.1017/9781108878715.026

Chapter 23 - DNA Damage: Fluorescent In-Situ Hybridization

Published online by Cambridge University Press: 05 April 2021

Edited by

Ralf Henkel and

Ashok Agarwal: Affiliation:
The Cleveland Clinic Foundation, Cleveland, OH
Ralf Henkel: Affiliation:
University of the Western Cape, South Africa
Ahmad Majzoub: Affiliation:
Hamad Medical Corporation, Doha

Book contents

Get access

Summary

Several sperm DNA integrity and chromatin quality assays have been developed and used for the detection of sperm DNA fragmentation and chromosomal aberrations [1, 2]. The Comet assay is one of the most commonly used single cell gel electrophoresis methods first developed by Östling and Johanson [3]. Damaged or fragmented sperm DNA is separated from undamaged DNA by electrophoresis. Fluorescent in situ hybridization (FISH) is a hybridization method that provides specific identification of selected DNA sequences and is frequently used for cytogenetic analysis of spermatozoa. These two methods were combined as the Comet-FISH technique first by Santos and colleagues in 1997 [4]. The Comet-FISH technique is a combination of two well-known methods, the Comet assay and FISH, and is a useful detection tool for screening of whole and region-specific DNA damage [5, 6]. In addition, the analysis of labeled DNA sequences and whole chromosomes of interest and identification of region-specific DNA and overall damage and repair in single cells is possible [7, 8]. Therefore, the modification of these assays interpolates hybridization with specific fluorescent-labeled probes to selected DNA sequences of interest after unwinding and electrophoresis [6] and developed a standardized Comet assay for human sperm with additional information for telomeres or special DNA sequences of interest. Thus, specific gene sequences can be detected using the Comet-FISH technique [9].

Type: Chapter
Information: Manual of Sperm Function Testing in Human Assisted Reproduction , pp. 228 - 233

DOI: https://doi.org/10.1017/9781108878715.026 [Opens in a new window]

Publisher: Cambridge University Press

Print publication year: 2021

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Book purchase

Temporarily unavailable

References

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(11): 1073–85. doi:10.1007/s10815-011-9631-8CrossRef Google Scholar PubMed

Simon, L, Liu, L, Murphy, K, Ge, S, Hotaling, J, Aston, KI, … Carrell, DT. Comparative analysis of three sperm DNA damage assays and sperm nuclear protein content in couples undergoing assisted reproduction treatment. Hum Reprod 2014; 29(5): 904–17. doi:10.1093/humrep/deu040CrossRef Google Scholar PubMed

Ostling, O, Johanson, KJ. Bleomycin, in contrast to gamma irradiation, induces extreme variation of DNA strand breakage from cell to cell. Int J Radiat Biol Relat Stud Phys Chem Med 1987; 52(5): 683–91. doi:10.1080/09553008714552201Google Scholar PubMed

Santos, SJ, Singh, NP, Natarajan, AT. Fluorescence in situ hybridization with comets. Exp Cell Res 1997; 232(2): 407–11. doi:10.1006/excr.1997.3555CrossRef Google Scholar PubMed

Glei, M, Hovhannisyan, G, Pool-Zobel, BL. Use of Comet-FISH in the study of DNA damage and repair: review. Mutat Res 2009; 681(1): 33–43. doi:10.1016/j.mrrev.2008.01.006CrossRef Google Scholar

Mondal, M, Guo, J. Comet-FISH for ultrasensitive strand-specific detection of DNA damage in single cells. Methods Enzymol 2017; 591: 83–95. doi:10.1016/bs.mie.2017.03.023CrossRef Google Scholar PubMed

Glei, M, Schlormann, W. Analysis of DNA damage and repair by comet fluorescence in situ hybridization (Comet-FISH). Methods Mol Biol 2014; 1094: 39–48. doi:10.1007/978-1-62703-706-8_4Google Scholar

Horvathova, E, Dusinska, M, Shaposhnikov, S, Collins, AR. DNA damage and repair measured in different genomic regions using the comet assay with fluorescent in situ hybridization. Mutagenesis 2004; 19(4): 269–76. doi:10.1093/mutage/geh030CrossRef Google Scholar PubMed

Shaposhnikov, S, El Yamani, N, Collins, AR. Fluorescent in situ hybridization on comets: FISH comet. Methods Mol Biol 2015; 1288: 363–73. doi:10.1007/978-1-4939-2474-5_21CrossRef Google Scholar PubMed

Hassold, T, Hunt, P. To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev Genet 2001; 2(4): 280–91. doi:10.1038/35066065Google Scholar

Kohn, TP, Kohn, JR, Darilek, S, Ramasamy, R, Lipshultz, L. Genetic counseling for men with recurrent pregnancy loss or recurrent implantation failure due to abnormal sperm chromosomal aneuploidy. J Assist Reprod Genet 2016; 33(5): 571–6. doi:10.1007/s10815-016-0702-8Google Scholar

Bernardini, L, Martini, E, Geraedts, JP, Hopman, AH, Lanteri, S, Conte, N, Capitanio, GL. Comparison of gonosomal aneuploidy in spermatozoa of normal fertile men and those with severe male factor detected by in-situ hybridization. Mol Hum Reprod 1997; 3(5): 431–8. doi:10.1093/molehr/3.5.431Google Scholar

Templado, C, Uroz, L, Estop, A. New insights on the origin and relevance of aneuploidy in human spermatozoa. Mol Hum Reprod 2013; 19(10): 634–43. doi:10.1093/molehr/gat039CrossRef Google Scholar PubMed

Colaco, S, Sakkas, D. Paternal factors contributing to embryo quality. J Assist Reprod Genet 2018; 35(11): 1953–68. doi:10.1007/s10815-018-1304-4CrossRef Google Scholar PubMed

Sarrate, Z, Anton, E. Fluorescence in situ hybridization (FISH) protocol in human sperm. J Vis Exp 2009; 31: 1405. doi:10.3791/1405Google Scholar

Blanco, J, Gabau, E, Gomez, D, Baena, N, Guitart, M, Egozcue, J, Vidal, F. Chromosome 21 disomy in the spermatozoa of the fathers of children with trisomy 21, in a population with a high prevalence of Down syndrome: increased incidence in cases of paternal origin. Am J Hum Genet 1998; 63(4): 1067–72. doi:10.1086/302058Google Scholar

Garcia-Mengual, E, Trivino, JC, Saez-Cuevas, A, Bataller, J, Ruiz-Jorro, M, Vendrell, X. Male infertility: establishing sperm aneuploidy thresholds in the laboratory. J Assist Reprod Genet 2019; 36(3): 371–81. doi:10.1007/s10815-018-1385-0Google Scholar

Sanchez-Castro, M, Jimenez-Macedo, AR, Sandalinas, M, Blanco, J. Prognostic value of sperm fluorescence in situ hybridization analysis over PGD. Hum Reprod 2009; 24(6): 1516–21. doi:10.1093/humrep/dep037Google Scholar

Ramasamy, R, Besada, S, Lamb, DJ. Fluorescent in situ hybridization of human sperm: diagnostics, indications, and therapeutic implications. Fertil Steril 2014; 102(6): 1534–9. doi:10.1016/j.fertnstert.2014.09.013Google Scholar

Rubio, C, Gil-Salom, M, Simon, C, Vidal, F, Rodrigo, L, Minguez, Y, … Pellicer, A. Incidence of sperm chromosomal abnormalities in a risk population: relationship with sperm quality and ICSI outcome. Hum Reprod 2001; 16(10): 2084–92. doi:10.1093/humrep/16.10.2084CrossRef Google Scholar

World Health Organization. (1999), WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction, 4th ed. Cambridge: Cambridge University Press.Google Scholar

Pang, MG, Hoegerman, SF, Cuticchia, AJ, Moon, SY, Doncel, GF, Acosta, AA, Kearns, WG. Detection of aneuploidy for chromosomes 4, 6, 7, 8, 9, 10, 11, 12, 13, 17, 18, 21, X and Y by fluorescence in-situ hybridization in spermatozoa from nine patients with oligoasthenoteratozoospermia undergoing intracytoplasmic sperm injection. Hum Reprod 1999; 14(5): 1266–73. doi:10.1093/humrep/14.5.1266Google Scholar

Caseiro, AL, Regalo, A, Pereira, E, Esteves, T, Fernandes, F, Carvalho, J. Implication of sperm chromosomal abnormalities in recurrent abortion and multiple implantation failure. Reprod Biomed Online 2015; 31(4): 481–5. doi:10.1016/j.rbmo.2015.07.001CrossRef Google Scholar PubMed

Bernardini, L, Gianaroli, L, Fortini, D, Conte, N, Magli, C, Cavani, S, … Venturini, PL. Frequency of hyper-, hypohaploidy and diploidy in ejaculate, epididymal and testicular germ cells of infertile patients. Hum Reprod 2000; 15(10): 2165–72. doi:10.1093/humrep/15.10.2165CrossRef Google Scholar PubMed

Burrello, N, Calogero, AE, De Palma, A, Grazioso, C, Torrisi, C, Barone, N, … Vicari, E. Chromosome analysis of epididymal and testicular spermatozoa in patients with azoospermia. Eur J Hum Genet 2002; 10(6): 362–6. doi:10.1038/sj.ejhg.5200814Google Scholar

Mateizel, I, Verheyen, G, Van Assche, E, Tournaye, H, Liebaers, I, Van Steirteghem, A. FISH analysis of chromosome X, Y and 18 abnormalities in testicular sperm from azoospermic patients. Hum Reprod 2002; 17(9): 2249–57. doi:10.1093/humrep/17.9.2249Google Scholar

Palermo, GD, Colombero, LT, Hariprashad, JJ, Schlegel, PN, Rosenwaks, Z. Chromosome analysis of epididymal and testicular sperm in azoospermic patients undergoing ICSI. Hum Reprod 2002; 17(3): 570–5. doi:10.1093/humrep/17.3.570CrossRef Google Scholar PubMed

Rodrigo, L, Rubio, C, Mateu, E, Simon, C, Remohi, J, Pellicer, A, Gil-Salom, M. Analysis of chromosomal abnormalities in testicular and epididymal spermatozoa from azoospermic ICSI patients by fluorescence in-situ hybridization. Hum Reprod 2004; 19(1): 118–23. doi:10.1093/humrep/deh012CrossRef Google Scholar PubMed

Gunes, S, Hekim, GN, Arslan, MA, Asci, R. Effects of aging on the male reproductive system. J Assist Reprod Genet 2016; 33(4): 441–54. doi:10.1007/s10815-016-0663-yCrossRef Google Scholar PubMed

Garcia-Ferreyra, J, Hilario, R, Luna, D, Villegas, L, Romero, R, Zavala, P, Duenas-Chacon, J. In vivo culture system using the INVOcell device shows similar pregnancy and implantation rates to those obtained from in vivo culture system in ICSI procedures. Clin Med Insights Reprod Health 2015; 9: 7–11. doi:10.4137/CMRH.S25494CrossRef Google Scholar PubMed

Garcia-Ferreyra, J, Luna, D, Villegas, L, Romero, R, Zavala, P, Hilario, R, Duenas-Chacon, J. High aneuploidy rates observed in embryos derived from donated oocytes are related to male aging and high percentages of sperm DNA fragmentation. Clin Med Insights Reprod Health 2015; 9: 21–7. doi:10.4137/CMRH.S32769CrossRef Google Scholar PubMed

Collodel, G, Moretti, E. Morphology and meiotic segregation in spermatozoa from men of proven fertility. J Androl 2008; 29(1): 106–14. doi:10.2164/jandrol.107.002998CrossRef Google Scholar PubMed

Mehdi, M, Gmidene, A, Brahem, S, Guerin, JF, Elghezal, H, Saad, A. Aneuploidy rate in spermatozoa of selected men with severe teratozoospermia. Andrologia 2012; 44(Suppl. 1): 139–43. doi:10.1111/j.1439-0272.2010.01152.xCrossRef Google Scholar PubMed

Calogero, AE, De Palma, A, Grazioso, C, Barone, N, Romeo, R, Rappazzo, G, D’Agata, R. Aneuploidy rate in spermatozoa of selected men with abnormal semen parameters. Hum Reprod 2001; 16(6): 1172–9. doi:10.1093/humrep/16.6.1172CrossRef Google Scholar PubMed

Lewis-Jones, I, Aziz, N, Seshadri, S, Douglas, A, Howard, P. Sperm chromosomal abnormalities are linked to sperm morphologic deformities. Fertil Steril 2003; 79(1): 212–15. doi:10.1016/s0015-0282(02)04411-4CrossRef Google Scholar PubMed

Tempest, HG, Griffin, DK. The relationship between male infertility and increased levels of sperm disomy. Cytogenet Genome Res 2004; 107(1–2): 83–94. doi:10.1159/000079575Google Scholar

Keymolen, K, Staessen, C, Verpoest, W, Liebaers, I, Bonduelle, M. Preimplantation genetic diagnosis in female and male carriers of reciprocal translocations: clinical outcome until delivery of 312 cycles. Eur J Hum Genet 2012; 20(4): 376–80. doi:10.1038/ejhg.2011.208Google Scholar

Munne, S, Sandalinas, M, Escudero, T, Fung, J, Gianaroli, L, Cohen, J. Outcome of preimplantation genetic diagnosis of translocations. Fertil Steril 2000; 73(6): 1209–18. doi:10.1016/s0015-0282(00)00495-7CrossRef Google Scholar PubMed

Tempest, HG. Meiotic recombination errors, the origin of sperm aneuploidy and clinical recommendations. Syst Biol Reprod Med 2011; 57(1–2): 93–101. doi:10.3109/19396368.2010.504879CrossRef Google Scholar PubMed

O’Flynn O’Brien, KL, Varghese, AC, Agarwal, A. The genetic causes of male factor infertility: a review. Fertil Steril 2010; 93(1): 1–12. doi:10.1016/j.fertnstert.2009.10.045CrossRef Google Scholar PubMed

Chatziparasidou, A, Christoforidis, N, Samolada, G, Nijs, M. Sperm aneuploidy in infertile male patients: a systematic review of the literature. Andrologia 2015; 47(8): 847–60. doi:10.1111/and.12362Google Scholar

Nicopoullos, JD, Gilling-Smith, C, Almeida, PA, Homa, S, Nice, L, Tempest, H, Ramsay, JW. The role of sperm aneuploidy as a predictor of the success of intracytoplasmic sperm injection? Hum Reprod 2008; 23(2): 240–50.Google Scholar PubMed