Skip to main content
SpringerLink
Log in

Ubiquitin-specific protease (USP26) gene alterations associated with male infertility and recurrent pregnancy loss (RPL) in Iranian infertile patients

  • Genetics
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Background and purpose

The human X chromosome is enriched with testis-specific genes that may be crucial for male fertility. Mutations in USP26 gene have been proposed to be associated with male infertility. Moreover, the importance of the ubiquitin pathway during different stages of mammalian fertilization and even embryo development has been addressed. Some mutations and haplotypes on this gene have been proposed to be associated with male infertility. In this study, five different mutations on USP26 were investigated: 1737 G > A, 1090 C > T, 370-371ins ACA, 494 T > C and 1423 C > T.

Methods

The study included 166 infertile men with non-obstructive azoospermia, 72 male partners of couples who had previously experienced ≥3 clinical first trimester spontaneous abortions and 60 fertile men. Besides family history of reproduction, hormonal evaluation and semen analysis were performed. DNA was extracted from blood samples. PCR-SSCP, PCR-RFLP and PCR Product Cloning methods were used and resumed by sequencing to insure about the mutations. Moreover, USP26 gene expression was studied by Real-Time PCR after RNA extraction followed by cDNA synthesis from 24 testis biopsies in obstructive and non-obstructive azoospermia patients.

Results

The results indicate that there is a haplotype between three observed mutations in Iranian population include: 370-371insACA, 1423C > T and 494 T > C. This haplotype was seen in control group as well. Surprisingly, total frequency of mutations in men with history of idiopathic RPL and azoospermic cases were significantly higher than that of in control groups (p < 0.05). Serum testosterone concentrations and testicular volume did not differ in the mutation positive group compared with the non-mutation group. About the USP26 gene expression, there is a significant difference between the expression levels of obstructive azoospermia, complete maturation arrest samples and SCO samples (P < 0.05).

Conclusions

According to our results, the USP26 gene may play an important role in male reproduction. The alterations of this gene may be involved in male infertility and RPL in Iranian population and may negatively affect testicular function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Zhang J, Qiu SD, Li SB, Zhou DX, Tian H, Huo YW, et al. Novel mutations in ubiquitin-specific protease 26 gene might cause spermatogenesis impairment and male infertility. Asian J Androl. 2007;9(6):809–14.

    Article  PubMed  CAS  Google Scholar 

  2. Huynh T, Mollard R, Trounson A. Selected genetic factors associated with male infertility. Hum Reprod Update. 2002;8(2):183–98.

    Article  PubMed  Google Scholar 

  3. Yang Y, Xiao CY, Zhou-Cun A, Zhang SZ, Li X, Zhang SX. DAZ1/DAZ2 cluster deletion mediated by gr/gr recombination per se may not be sufficient for spermatogenesis impairment: a study of Chinese normozoospermic men. Asian J Androl. 2006;8(2):183–7.

    Article  PubMed  CAS  Google Scholar 

  4. Totonchi M, Mohseni Meybodi A, Borjian Boroujeni P, Sedighi Gilani M, Almadani N, Gourabi H. Clinical data for 185 infertile Iranian men with Y-chromosome microdeletion. J Assist Reprod Genet. 2012;29(8):847–53.

    Article  PubMed  Google Scholar 

  5. Fernando L, Gromoll J, Weerasooriya TR, Nieschlag E, Simoni M. Y-chromosomal microdeletions and partial deletions of the Azoospermia Factor c (AZFc) region in normozoospermic, severe oligozoospermic and azoospermic men in Sri Lanka. Asian J Androl. 2006;8(1):39–44.

    Article  PubMed  CAS  Google Scholar 

  6. Stouffs K, Lissens W, Tournaye H, Van Steirteghem A, Liebaers I. Possible role of USP26 in patients with severely impaired spermatogenesis. Eur J Hum Genet. 2005;13(3):336–40.

    Article  PubMed  CAS  Google Scholar 

  7. Stouffs K, Willems A, Lissens W, Tournaye H, Van Steirteghem A, Liebaers I. The role of the testis-specific gene hTAF7L in the aetiology of male infertility. Mol Hum Reprod. 2006;12(4):263–7.

    Article  PubMed  CAS  Google Scholar 

  8. Nishimune Y, Tanaka H. Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes. J Androl. 2006;27(3):326–34.

    Article  PubMed  CAS  Google Scholar 

  9. Wang PJ, McCarrey JR, Yang F, Page DC. An abundance of X-linked genes expressed in spermatogonia. Nat Genet. 2001;27(4):422–6.

    Article  PubMed  Google Scholar 

  10. Glickman MH, Ciechanover A. The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. Physiol Rev. 2002;82(2):373–428.

    PubMed  CAS  Google Scholar 

  11. Amerik AY, Hochstrasser M. Mechanism and function of deubiquitinating enzymes. Biochim Biophys Acta. 2004;1695(1–3):189–207.

    Article  PubMed  CAS  Google Scholar 

  12. Wilkinson KD. Regulation of ubiquitin-dependent processes by deubiquitinating enzymes. FASEB J. 1997;11(14):1245–56.

    PubMed  CAS  Google Scholar 

  13. Lee IW, Kuan LC, Lin CH, Pan HA, Hsu CC, Tsai YC, et al. Association of USP26 haplotypes in men in Taiwan, China with severe spermatogenic defect. Asian J Androl. 2008;10(6):896–904.

    Article  PubMed  CAS  Google Scholar 

  14. Wright A, Reiley WW, Chang M, Jin W, Lee AJ, Zhang M, et al. Regulation of early wave of germ cell apoptosis and spermatogenesis by deubiquitinating enzyme CYLD. Dev Cell. 2007;13(5):705–16.

    Article  PubMed  CAS  Google Scholar 

  15. Christensen GL, Griffin J, Carrell DT. Sequence analysis of the X-linked USP26 gene in severe male factor infertility patients and fertile controls. Fertil Steril. 2008;90(3):851–2.

    Article  PubMed  CAS  Google Scholar 

  16. Nijman SM, Luna-Vargas MP, Velds A, Brummelkamp TR, Dirac AM, Sixma TK, et al. A genomic and functional inventory of deubiquitinating enzymes. Cell. 2005;123(5):773–86.

    Article  PubMed  CAS  Google Scholar 

  17. Sakai N, Sawada MT, Sawada H. Non-traditional roles of ubiquitin-proteasome system in fertilization and gametogenesis. Int J Biochem Cell Biol. 2004;36(5):776–84.

    Article  PubMed  CAS  Google Scholar 

  18. Baarends WM, Roest HP, Grootegoed JA. The ubiquitin system in gametogenesis. Mol Cell Endocrinol. 1999;151(1–2):5–16.

    Article  PubMed  CAS  Google Scholar 

  19. Baarends WM, van der Laan R, Grootegoed JA. Specific aspects of the ubiquitin system in spermatogenesis. J Endocrinol Invest. 2000;23(9):597–604.

    PubMed  CAS  Google Scholar 

  20. Ford HB, Schust DJ. Recurrent pregnancy loss: etiology, diagnosis, and therapy. Rev Obstet Gynecol. 2009;2(2):76–83.

    PubMed  Google Scholar 

  21. Porter TF, Scott JR. Evidence-based care of recurrent miscarriage. Best Pract Res Clin Obstet Gynaecol. 2005;19(1):85–101.

    Article  PubMed  Google Scholar 

  22. Mozdarani H, Meybodi AM, Zari-Moradi S. A cytogenetic study of couples with recurrent spontaneous abortions and infertile patients with recurrent IVF/ICSI failure. Indian J Hum Genet. 2008;14(1):1–6.

    Article  PubMed  Google Scholar 

  23. Franssen MT, Korevaar JC, van der Veen F, Leschot NJ, Bossuyt PM, Goddijn M. Reproductive outcome after chromosome analysis in couples with two or more miscarriages: index [corrected]-control study. BMJ. 2006;332(7544):759–63.

    Article  PubMed  Google Scholar 

  24. Franssen MT, Musters AM, van der Veen F, Repping S, Leschot NJ, Bossuyt PM, et al. Reproductive outcome after PGD in couples with recurrent miscarriage carrying a structural chromosome abnormality: a systematic review. Hum Reprod Update. 2011;17(4):467–75.

    Article  PubMed  CAS  Google Scholar 

  25. 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(7):1713–21.

    Article  PubMed  CAS  Google Scholar 

  26. Borini A, Tarozzi N, Bizzaro D, Bonu MA, Fava L, Flamigni C, et al. Sperm DNA fragmentation: paternal effect on early post-implantation embryo development in ART. Hum Reprod. 2006;21(11):2876–81.

    Article  PubMed  CAS  Google Scholar 

  27. Kennedy C, Ahlering P, Rodriguez H, Levy S, Sutovsky P. Sperm chromatin structure correlates with spontaneous abortion and multiple pregnancy rates in assisted reproduction. Reprod Biomed Online. 2011;22(3):272–6.

    Article  PubMed  CAS  Google Scholar 

  28. Lewis SE, Agbaje I, Alvarez J. Sperm DNA tests as useful adjuncts to semen analysis. Syst Biol Reprod Med. 2008;54(3):111–25.

    Article  PubMed  CAS  Google Scholar 

  29. Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM. Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril. 2008;90(2):352–9.

    Article  PubMed  Google Scholar 

  30. Zini A, Boman JM, Belzile E, Ciampi A. Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis. Hum Reprod. 2008;23(12):2663–8.

    Article  PubMed  CAS  Google Scholar 

  31. Zini A. Are sperm chromatin and DNA defects relevant in the clinic? Syst Biol Reprod Med. 2011;57(1–2):78–85.

    Article  PubMed  Google Scholar 

  32. Collins JA, Barnhart KT, Schlegel PN. Do sperm DNA integrity tests predict pregnancy with in vitro fertilization? Fertil Steril. 2008;89(4):823–31.

    Article  PubMed  Google Scholar 

  33. Nicopoullos JD, Gilling-Smith C, Almeida PA, Homa S, Norman-Taylor JQ, Ramsay JW. Sperm DNA fragmentation in subfertile men: the effect on the outcome of intracytoplasmic sperm injection and correlation with sperm variables. BJU Int. 2008;101(12):1553–60.

    Article  PubMed  Google Scholar 

  34. Tavalaee M, Razavi S, Nasr-Esfahani MH. Influence of sperm chromatin anomalies on assisted reproductive technology outcome. Fertil Steril. 2009;91(4):1119–26.

    Article  PubMed  CAS  Google Scholar 

  35. Baarends WM, van der Laan R, Grootegoed JA. DNA repair mechanisms and gametogenesis. Reproduction. 2001;121(1):31–9.

    Article  PubMed  CAS  Google Scholar 

  36. Shamsi MB, Kumar K, Dada R. Genetic and epigenetic factors: role in male infertility. Indian J Urol. 2011;27(1):110–20.

    Article  PubMed  CAS  Google Scholar 

  37. Bebington C, Doherty FJ, Fleming SD. Ubiquitin and ubiquitin-protein conjugates are present in human cytotrophoblast throughout gestation. Early Pregnancy. 2000;4(4):240–52.

    PubMed  CAS  Google Scholar 

  38. Yi YJ, Manandhar G, Sutovsky M, Li R, Jonakova V, Oko R, et al. Ubiquitin C-terminal hydrolase-activity is involved in sperm acrosomal function and anti-polyspermy defense during porcine fertilization. Biol Reprod. 2007;77(5):780–93.

    Article  PubMed  CAS  Google Scholar 

  39. Yi YJ, Manandhar G, Oko RJ, Breed WG, Sutovsky P. Mechanism of sperm-zona pellucida penetration during mammalian fertilization: 26S proteasome as a candidate egg coat lysin. Soc Reprod Fertil Suppl. 2007;63:385–408.

    PubMed  CAS  Google Scholar 

  40. Paduch DA, Mielnik A, Schlegel PN. Novel mutations in testis-specific ubiquitin protease 26 gene may cause male infertility and hypogonadism. Reprod Biomed Online. 2005;10(6):747–54.

    Article  PubMed  Google Scholar 

  41. Ribarski I, Lehavi O, Yogev L, Hauser R, Bar-Shira Maymon B, Botchan A, et al. USP26 gene variations in fertile and infertile men. Hum Reprod. 2009;24(2):477–84.

    Article  PubMed  CAS  Google Scholar 

  42. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988;16(3):1215.

    Article  PubMed  CAS  Google Scholar 

  43. Vogt PH. Molecular genetics of human male infertility: from genes to new therapeutic perspectives. Curr Pharm Des. 2004;10(5):471–500.

    Article  PubMed  CAS  Google Scholar 

  44. Zarei-Kheirabadi M, Shayegan Nia E, Tavalaee M, Deemeh MR, Arabi M, Forouzanfar M, et al. Evaluation of ubiquitin and annexin V in sperm population selected based on density gradient centrifugation and zeta potential (DGC-Zeta). J Assist Reprod Genet. 2012;29(4):365–71.

    Article  PubMed  CAS  Google Scholar 

  45. Miyamoto T, Tsujimura A, Miyagawa Y, Koh E, Namiki M, Horikawa M, et al. Single nucleotide polymorphism in the UBR2 gene may be a genetic risk factor for Japanese patients with azoospermia by meiotic arrest. J Assist Reprod Genet. 2011;28(8):743–6.

    Article  PubMed  Google Scholar 

  46. Koslowski M, Sahin U, Huber C, Tureci O. The human X chromosome is enriched for germline genes expressed in premeiotic germ cells of both sexes. Hum Mol Genet. 2006;15(15):2392–9.

    Article  PubMed  CAS  Google Scholar 

  47. Ravel C, El Houate B, Chantot S, Lourenco D, Dumaine A, Rouba H, et al. Haplotypes, mutations and male fertility: the story of the testis-specific ubiquitin protease USP26. Mol Hum Reprod. 2006;12(10):643–6.

    Article  PubMed  CAS  Google Scholar 

  48. Stouffs K, Lissens W, Tournaye H, Van Steirteghem A, Liebaers I. Alterations of the USP26 gene in Caucasian men. Int J Androl. 2006;29(6):614–7.

    Article  PubMed  CAS  Google Scholar 

  49. Tuttelmann F, Rajpert-De Meyts E, Nieschlag E, Simoni M. Gene polymorphisms and male infertility—a meta-analysis and literature review. Reprod Biomed Online. 2007;15(6):643–58.

    Article  PubMed  Google Scholar 

  50. Shamsi MB, Venkatesh S, Tanwar M, Singh G, Mukherjee S, Malhotra N, et al. Comet assay: a prognostic tool for DNA integrity assessment in infertile men opting for assisted reproduction. Indian J Med Res. 2010;131:675–81.

    PubMed  CAS  Google Scholar 

  51. 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(2):115–23.

    PubMed  CAS  Google Scholar 

  52. Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update. 2003;9(4):331–45.

    Article  PubMed  CAS  Google Scholar 

  53. Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl. 2002;23(1):25–43.

    PubMed  Google Scholar 

Download references

Acknowledgments

We express gratitude to all the participants involved in this study. We acknowledge the efforts of genetic laboratory staff of Royan Reproductive biomedicine research center specially Mrs. Anissi and Mrs Mokhtari.

Declaration of interest

The authors declare that no conflicts of interest exist. This work was supported by Royan Institute for Reproductive biomedicine, ACECR.

Author information

Authors and Affiliations

  1. Department of Genetics at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    U. Asadpor, M. Totonchi, H. Hoseinifar, Sh. Zari Moradi, H. Gourabi & A. Mohseni Meybodi

  2. Department of Andrology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    M. Sabbaghian & M. A. Sadighi Gilani

  3. Statistics Department, Mathematical Science and computer Faculty, Shahid Chamran University, Ahwaz, Iran

    M. R. Akhound

  4. Department of Pathobiology, Faculty of Medicine, Alborz University of Medical Science, Karaj, Iran

    K. Haratian

Authors
  1. U. Asadpor
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Totonchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Sabbaghian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. Hoseinifar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. R. Akhound
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sh. Zari Moradi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. K. Haratian
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M. A. Sadighi Gilani
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. H. Gourabi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. A. Mohseni Meybodi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. Mohseni Meybodi.

Additional information

Capsule In recent years, a lot of attention has been paid to genetic causes of male infertility. Moreover altered gene expression in spermatogenesis has been showed. Studies are mainly focusing on genes with a testis-specific expression pattern. Such genes which located on the sex chromosomes seem to be more important as men are hemizygous for these chromosomes. We analyzed the Ubiquitin Specific Protease 26 (USP26) gene for the presence of mutations in men with severe fertility problems.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Asadpor, U., Totonchi, M., Sabbaghian, M. et al. Ubiquitin-specific protease (USP26) gene alterations associated with male infertility and recurrent pregnancy loss (RPL) in Iranian infertile patients. J Assist Reprod Genet 30, 923–931 (2013). https://doi.org/10.1007/s10815-013-0027-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-013-0027-9

Keywords

Search

Navigation