Recent studies have shown that infertility affects estimated 15% of all couples. Male infertility is the primary or contributory cause in 60% of these cases. Consequently, the application of assisted reproduction is increasing. These methods could benefit from an extended evaluation of sperm quality. For this reason, we analyzed sperm proteins from 30 men with normal spermiograms and 30 men with asthenozoospermia. Ejaculates of both groups were tested by flow cytometry (FCM) and fluorescence with a set of well-characterized anti-human sperm Hs-monoclonal antibodies (MoAbs), which were generated in our laboratory. No statistically significant differences were found between normospermics and asthenospermics in the expression of the sperm surface protein clusterin, evaluated with Hs-3 MoAb, and semenogelin, evaluated with Hs-9 MoAb. However, FCM revealed quantitative differences in the acrosomal proteins between normozoospermic and asthenozoospermic men, namely, in glyceraldehyde-3-phosphate dehydrogenase, evaluated with Hs-8 MoAb, valosin-containing protein, evaluated with Hs-14 MoAb, and ATP synthase (cAMP-dependent protein kinase II, PRKAR2A), evaluated with MoAb Hs-36. Asthenozoospermic men displayed a highly reduced expression of intra-acrosomal proteins, with a likely decrease in sperm quality, and thus a negative impact on successful reproduction. Asthenozoospermia seems to be a complex disorder involving intra-acrosomal proteins.

Laboratory of Reproductive Biology Institute of Biotechnology Academy of Sciences of the Czech Republic v v i Prague Czech Republic

Zobrazit více v PubMed

5th ed. Geneva, Switzerland: World Health Organization; 2010. World Health Organization. WHO Laboratory Manual for the Examination and Processing of human semen.

Oliva R, Martínez-Heredia J, Estanyol JM. Proteomics in the study of the sperm cell composition, differentiation and function. Syst Biol Reprod Med. 2008;54:23–36. PubMed

Duplessis SS, Kashou AH, Benjamin DJ, Yadav SP, Agarval A. Proteomics: a subcellular look at spermatozoa. Reprod Biol Endocrinol. 2011;9:36–47. PubMed PMC

Baker MA, Nixon B, Naumovski N, Aitken RJ. Proteomic insights into the maturation and capacitation of mammalian spermatozoa. Syst Biol Reprod Med. 2012;58:211–7. PubMed

Yoshiki T, Lai BC, Dorjee S, Lee CY. Molecular characterizations of an intraacrosomal antigen defined by HS-33 monoclonal antibody. J Androl. 1996;17:666–73. PubMed

Dorjee S, Lai CL, Lee A, Lee CY. Monoclonal antibodies as direct probes for human sperm acrosome reaction. Am J Reprod Immunol. 1997;37:283–90. PubMed

Hamatani T, Tanabe K, Kamei K, Sakai N, Yamamoto Y, et al. A monoclonal antibody to human SP-10 inhibits in vitro the binding of human sperm to hamster oolemma but not to human Zona pellucida. Biol Reprod. 2000;62:1201–8. PubMed

Naz RK, Chauhan SC, Trivedi RN. Monoclonal antibody against human sperm-specific YLP12 peptide sequence involved in oocyte binding. Arch Androl. 2002;48:169–75. PubMed

Reddy KV, Vijayalaxmi G, Rajeev KS, Aranha C. Inhibition of sperm-egg binding and fertilisation in mice by a monoclonal antibody reactive to 57-kDa human sperm surface antigen. Reprod Fertil Dev. 2006;18:875–84. PubMed

Ulcova-Gallova Z, Gruberova J, Vrzalova J, Bibkova K, Peknicova J, et al. Sperm antibodies, intra-acrosomal sperm proteins, and cytokines in semen in men from infertile couples. Am J Reprod Immunol. 2009;61:236–45. PubMed

Veaute C, Liu de Y, Furlong LI, Biancotti JC, Baker HW, et al. Anti-human proacrosin antibody inhibits the zona pellucida (ZP)-induced acrosome reaction of ZP-bound spermatozoa. Fertil Steril. 2010;93:2456–9. PubMed

Capkova J, Geussova G, Peknicova J. Monoclonal antibody to human sperm acrosomal protein. Folia Biol (Praha) 2000;46:55–7. PubMed

Capková J, Geussová G, Peknicová J. New monoclonal antibody to human apolipoprotein J. Folia Biol (Praha) 2002;48:40–2. PubMed

Margaryan H. MAb Hs-9 against human semenogelin. Hybridoma. 2006;25:254.

Capkova J, Margaryan H, Elzeinova F, Koubek P, Peknicova J. Monoclonal antibodies to human seminal plasma proteins. In: Lagarda JL, Oliva R, editors. Proceedings of the 9th International Congress of Andrology. Barcelona (Spain): MEDIMOND; 2009. pp. 95–101.

Margaryan H, Peknicova J, Capkova J. Characterization of Human Sperm Protein Recognized by Monoclonal Antibody Hs-8. Proceedings of the 13th International Symposium for Immunology of Reproduction, Varna (Bulgaria) 2012:27.

Cordelli E, Eleuteri P, Leter G, Rescia M, Spanò M. Flow cytometry applications in the evaluation of sperm quality: semen analysis, sperm function and DNA integrity. Contraception. 2005;72:273–9. PubMed

Mahfouz RZ, Tamer MS, Argaval A. The diagnostic and therapeutic applications of flow cytometry in male infertility. Arch Med Sci. 2009;5(1A):S99–108.

Robles V, Martínez-Pastor F. Flow cytometric methods for sperm assessment. Methods Mol Biol. 2013;927:175–86. PubMed

Petrunkina AM, Harrison RA. Fluorescence technologies for evaluating male gamete (dys) function. Reprod Domest Anim. 2013;48(Suppl 1):11–24. PubMed

Homolka D, Ivanek R, Capkova J, Jansa P, Forejt J. Chromosomal rearrangement interferes with meiotic X chromosome inactivation. Genome Res. 2007;17:1431–7. PubMed PMC

Ded L, Dostalova P, Dorosh A, Dvorakova-Hortova K, Peknicova J. Effect of estrogens on boar sperm capacitation in vitro. Reprod Biol Endocrinol. 2010;8:87. PubMed PMC

Peknicova J, Chladek D, Hozak P. Monoclonal antibodies against sperm intra-acrosomal antigens as markers for male infertility diagnostics and estimation of spermatogenesis. Am J Reprod Immunol. 2005;53:42–9. PubMed

Pixton KL, Deeks ED, Flesch FM, Moseley FL, Björndahl L, et al. Sperm proteome mapping of a patient who experienced failed fertilization at IVF reveals altered expression of at least 20 proteins compared with fertile donors: case report. Hum Reprod. 2004;19:1438–47. PubMed

Liao TT, Xiang Z, Zhu WB, Fan LQ. Proteome analysis of round-headed and normal spermatozoa by 2-D fluorescence difference gel electrophoresis and mass spectrometry. Asian J Androl. 2009;11:683–93. PubMed PMC

Thacker S, Yadav SP, Sharma RK, Kashou A, Willard B, et al. Evaluation of sperm proteins in infertile men: a proteomic approach. Fertil Steril. 2011;95:2745–8. PubMed

Koide SS, Wang L, Kamada M. Antisperm antibodies associated with infertility: properties and encoding genes of target antigens. Proc Soc Exp Biol Med. 2000;224:123–32. PubMed

Hasegawa A, Fu Y, Tsubamoto H, Tsuji Y, Sawai H, et al. Epitope analysis for human sperm-immobilizing monoclonal antibodies, MAb H6-3C4, 1G12 and campath-1. Mol Hum Reprod. 2003;9:337–43. PubMed

Peknicova J, Pexidrova M, Kubatova A, Koubek P, Tepla O, et al. Expression of beta-tubulin epitope in human sperm with pathological spermiogram. Fertil Steril. 2007;88:1120–8. PubMed

Tepla O, Peknicova J, Koci K, Mika J, Mrazek M, et al. Evaluation of reproductive potential after intracytoplasmic sperm injection of varied human semen tested by antiacrosomal antibodies. Fertil Steril. 2006;86:113–20. PubMed

Eustache F, Jouannet P, Auger J. Evaluation of flow cytometric methods to measure human sperm concentration. J Androl. 2001;22:558–67. PubMed

Graham JK, Kunze E, Hammerstedt RH. Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry. Biol Reprod. 1990;43:55–64. PubMed

Tao J, Du J, Critser ES, Critser JK. Assessment of the acrosomal status and viability of human spermatozoa simultaneously using flow cytometry. Hum Reprod. 1993;8:1879–85. PubMed

Zoppino FC, Halón ND, Bustos MA, Pavarotti MA, Mayorga LS. Recording and sorting live human sperm undergoing acrosome reaction. Fertil Steril. 2012;97:1309–15. PubMed

Auger J, Leonce S, Jouannet P, Ronot X. Flow cytometric sorting of living, highly motile human spermatozoa based on evaluation of their mitochondrial activity. J Histochem Cytochem. 1993;41:1247–51. PubMed

Evenson D, Jost L. Sperm chromatin structure assay is useful for fertility assessment. Methods Cell Sci. 2000;22:169–89. PubMed

Zini A, Kamal K, Phang D, Willis J, Jarvi K. Biologic variability of sperm DNA denaturation in infertile men. Urology. 2001;58:258–61. PubMed

Venkatesh S, Singh A, Shamsi MB, Thilagavathi J, Kumar R, et al. Clinical significance of sperm DNA damage threshold value in the assessment of male infertility. Reprod Sci. 2011;18:1005–13. PubMed

Ribeiro SC, Sartorius G, Pletscher F, de Geyter M, Zhang H, et al. Isolation of spermatozoa with low levels of fragmented DNA with the use of flow cytometry and sorting. Fertil Steril. 2013;100:686–94. PubMed

Aslam I, Robins A, Dowell K, Fishel S. Isolation, purification and assessment of viability of spermatogenic cells from testicular biopsies of azoospermic men. Hum Reprod. 1998;13:639–45. PubMed

Piasecka M, Gaczarzewicz D, Laszczynska M, Starczewski A, Brodowska A. Flow cytometry application in the assessment of sperm DNA integrity of men with asthenozoospermia. Folia Histochem Cytobiol. 2007;45(Suppl 1):S127–36. PubMed

Capková J, Elzeinová F, Novák P. Increased expression of secretory actin-binding protein on human spermatozoa is associated with poor semen quality. Hum Reprod. 2007;22:1396–404. PubMed

Zatecka E, Ded L, Elzeinova F, Kubatova A, Dorosh A, et al. Effect of tetrabrombisphenol A on induction of apoptosis in the testes and changes in expression of selected testicular genes in CD1 mice. Reprod Toxicol. 2013;35:32–9. PubMed

Comprehensive Flow-Cytometric Quality Assessment of Ram Sperm Intended for Gene Banking Using Standard and Novel Fertility Biomarkers

The Transgenerational Transmission of the Paternal Type 2 Diabetes-Induced Subfertility Phenotype