Advertisement

Journal of Assisted Reproduction and Genetics

, Volume 30, Issue 9, pp 1125–1131 | Cite as

Lack of association of MTHFR rs1801133 polymorphism and CTCFL mutations with sperm methylation errors in infertile patients

  • Cristina Camprubí
  • Marta Pladevall
  • Mark Grossmann
  • Nicolás Garrido
  • Maria C. Pons
  • Joan Blanco
Genetics

Abstract

Purpose

To find out whether the MTHFR rs1801133 polymorphism is a risk factor for male infertility in the Spanish population. To determine if a pattern of sperm DNA hypomethylation at the paternally imprinted loci H19-ICR and/or IG-DMR is related to the MTHFR rs1801133 polymorphism and/or CTCFL mutations.

Methods

One hundred and seven samples from individuals who sought consultation for fertility problems and twenty-five semen samples from sperm donors were analyzed. The MTHFR rs1801133 SNP was analyzed in all samples by the PCR-RFLP method. We compared the distribution of the genotypes between control and infertile populations and among the groups of patients with altered seminal parameters. In those patients with the most severe hypomethylation pattern (n = 12) we also analyzed the CTCFL protein-coding exons by sequencing.

Results

There were no significant differences in the distribution of the genotypes among the control and infertile populations. Moreover, none of the genotypes were associated, neither to the characteristics of the seminogram, nor to the presence of sperm DNA hypomethylation. We did not identify frameshift, nonsense or missense mutations of the CTCFL gene.

Conclusions

The MTHFR rs1801133 polymorphism is not associated with male infertility in the Spanish population. Neither the MTHFR polymorphism, nor CTCFL mutations explain a pattern of sperm hypomethylation at paternally imprinting loci.

Keywords

MTHFR polymorphism CTCFL DNA methylation Male infertility Imprinting errors 

Notes

Acknowledgments

The authors wish to thank Dr. Javier Nadal and the embryologist from the Unidad de Reproducción Asistida of the Centro Médico Teknon (Barcelona, Spain) and the Laboratorio de Andrología y Banco de Semen of the Instituto Universitario IVI Valencia (Valencia, Spain) for providing the semen samples. This work was supported by Projects PS09/00330 (Fondo de Investigación Sanitaria, Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, Spain) and SGR2009–282 (Agència de Gestió d’Ajuts Universitaris i de Recerca, Generalitat de Catalunya, Spain). Marta Pladevall was the recipient of the grant number UAB2006–00213 from the Universitat Autònoma de Barcelona.

Supplementary material

10815_2013_13_MOESM1_ESM.doc (176 kb)
Supplemental Table 1 Age and spermiogram parameters of the infertile patients. (DOC 175 kb)

References

  1. 1.
    A ZC, Yang Y, Zhang SZ, Li N, Zhang W. Single nucleotide polymorphism C677T in the methylenetetrahydrofolate reductase gene might be a genetic risk factor for infertility for Chinese men with azoospermia or severe oligozoospermia. Asian J Androl. 2007;9:57–62.PubMedCrossRefGoogle Scholar
  2. 2.
    Aapola U, Kawasaki K, Scott HS, Ollila J, Vihinen M, Heino M, et al. Isolation and initial characterization of a novel zinc finger gene, DNMT3L, on 21q22.3, related to the cytosine-5-methyltransferase 3 gene family. Genomics. 2000;65:293–8.PubMedCrossRefGoogle Scholar
  3. 3.
    Bezold G, Lange M, Peter RU. Homozygous methylenetetrahydrofolate reductase C677T mutation and male infertility. New Engl J Med. 2001;344:1172–3.PubMedCrossRefGoogle Scholar
  4. 4.
    Boissonnas CC, Abdalaoui HE, Haelewyn V, Fauque P, Dupont JM, Gut I, et al. Specific epigenetic alterations of IGF2-H19 locus in spermatozoa from infertile men. Eur J Hum Genet. 2010;18:73–80.PubMedCrossRefGoogle Scholar
  5. 5.
    Camprubí C, Pladevall M, Grossmann M, Garrido N, Pons M, Blanco J. Semen samples showing an increased rate of spermatozoa with imprinting errors have a negligible effect in the outcome of assisted reproduction techniques. Epigenetics. 2012;7:1115–1124.Google Scholar
  6. 6.
    Dhillon VS, Shahid M, Husain SA. Associations of MTHFR DNMT3b 4977 bp deletion in mtDNA and GSTM1 deletion, and aberrant CpG island hypermethylation of GSTM1 in non-obstructive infertility in Indian men. Mol Hum Reprod. 2007;13:213–22.PubMedCrossRefGoogle Scholar
  7. 7.
    Ebisch IM, van Heerde WL, Thomas CM, van der Put N, Wong WY, Steegers-Theunissen RP. C677T methylenetetrahydrofolate reductase polymorphism interferes with the effects of folic acid and zinc sulfate on sperm concentration. Fertil Steril. 2003;80:1190–4.PubMedCrossRefGoogle Scholar
  8. 8.
    El Hajj N, Zechner U, Schneider E, Tresch A, Gromoll J, Hahn T, et al. Methylation status of imprinted genes and repetitive elements in sperm DNA from infertile males. Sex Dev. 2011;5:60–9.PubMedCrossRefGoogle Scholar
  9. 9.
    Frosst P, Blom HJ, Milos R, Goyette P, Sheppard CA, Matthews RG, et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet. 1995;10:111–3.PubMedCrossRefGoogle Scholar
  10. 10.
    Gava MM, Chagas Ede O, Bianco B, Christofolini DM, Pompeo AC, Glina S, et al. Methylenetetrahydrofolate reductase polymorphisms are related to male infertility in Brazilian men. Genet Test Mol Biomarkers. 2011;15:153–7.PubMedCrossRefGoogle Scholar
  11. 11.
    Gupta N, Gupta S, Dama M, David A, Khanna G, Khanna A, et al. Strong association of 677 C > T substitution in the MTHFR gene with male infertility—a study on an indian population and a meta-analysis. PLoS One. 2011;6:e22277.PubMedCrossRefGoogle Scholar
  12. 12.
    Hammoud SS, Purwar J, Pflueger C, Cairns BR, Carrell DT. Alterations in sperm DNA methylation patterns at imprinted loci in two classes of infertility. Fertil Steril. 2010;94:1728–33.PubMedCrossRefGoogle Scholar
  13. 13.
    Jelinic P, Stehle JC, Shaw P. The testis-specific factor CTCFL cooperates with the protein methyltransferase PRMT7 in H19 imprinting control region methylation. PLoS Biol. 2006;4:e355.PubMedCrossRefGoogle Scholar
  14. 14.
    Kobayashi H, Hiura H, John RM, Sato A, Otsu E, Kobayashi N, et al. DNA methylation errors at imprinted loci after assisted conception originate in the parental sperm. Eur J Hum Genet. 2009;17:1582–91.PubMedCrossRefGoogle Scholar
  15. 15.
    Kobayashi H, Sato A, Otsu E, Hiura H, Tomatsu C, Utsunomiya T, et al. Aberrant DNA methylation of imprinted loci in sperm from oligospermic patients. Hum Mol Genet. 2007;16:2542–51.PubMedCrossRefGoogle Scholar
  16. 16.
    Lee HC, Jeong YM, Lee SH, Cha KY, Song SH, Kim NK, et al. Association study of four polymorphisms in three folate-related enzyme genes with non-obstructive male infertility. Hum Reprod. 2006;21:3162–70.PubMedCrossRefGoogle Scholar
  17. 17.
    Marques CJ, Carvalho F, Sousa M, Barros A. Genomic imprinting in disruptive spermatogenesis. Lancet. 2004;363:1700–2.PubMedCrossRefGoogle Scholar
  18. 18.
    Marques CJ, Costa P, Vaz B, Carvalho F, Fernandes S, Barros A, et al. Abnormal methylation of imprinted genes in human sperm is associated with oligozoospermia. Mol Hum Reprod. 2008;14:67–74.PubMedCrossRefGoogle Scholar
  19. 19.
    Marques CJ, Francisco T, Sousa S, Carvalho F, Barros A, Sousa M. Methylation defects of imprinted genes in human testicular spermatozoa. Fertil Steril. 2010;94:585–94.PubMedCrossRefGoogle Scholar
  20. 20.
    Minor A, Chow V, Ma S. Aberrant DNA methylation at imprinted genes in testicular sperm retrieved from men with obstructive azoospermia and undergoing vasectomy reversal. Reproduction. 2011;141:749–57.PubMedCrossRefGoogle Scholar
  21. 21.
    Paracchini V, Garte S, Taioli E. MTHFR C677T polymorphism, GSTM1 deletion and male infertility: a possible suggestion of a gene-gene interaction? Biomarkers. 2006;11:53–60.PubMedCrossRefGoogle Scholar
  22. 22.
    Park JH, Lee HC, Jeong YM, Chung TG, Kim HJ, Kim NK, et al. MTHFR C677T polymorphism associates with unexplained infertile male factors. J Assist Reprod Genet. 2005;22:361–8.PubMedCrossRefGoogle Scholar
  23. 23.
    Poplinski A, Tüttelmann F, Kanber D, Horsthemke B, Gromoll J. Idiopathic male infertility is strongly associated with aberrant methylation of MEST and IGF2/H19 ICR1. Int J Androl. 2010;33:642–9.PubMedGoogle Scholar
  24. 24.
    Ravel C, Chantot-Bastaraud S, Chalmey C, Barreiro L, Aknin-Seifer I, Pfeffer J, et al. Lack of association between genetic polymorphisms in enzymes associated with folate metabolism and unexplained reduced sperm counts. PLoS One. 2009;4:e6540.PubMedCrossRefGoogle Scholar
  25. 25.
    Safarinejad MR, Shafiei N, Safarinejad S. Relationship between genetic polymorphisms of methylenetetrahydrofolate reductase (C677T, A1298C, and G1793A) as risk factors for idiopathic male infertility. Reprod Sci. 2011;18:304–15.PubMedCrossRefGoogle Scholar
  26. 26.
    Singh K, Singh SK, Sah R, Singh I, Raman R. Mutation C677T in the methylenetetrahydrofolate reductase gene is associated with male infertility in an Indian population. Int J Androl. 2005;28:115–9.PubMedCrossRefGoogle Scholar
  27. 27.
    Solvas I, Grossmann M, Santaló J, Pons MC. Estudio comparativo entre dos métodos de swim-up. Revista ASEBIR. 2002;7:28–32.Google Scholar
  28. 28.
    Stuppia L, Gatta V, Scarciolla O, Colosimo A, Guanciali-Franchi P, Calabrese G, et al. The methylenetethrahydrofolate reductase (MTHFR) C677T polymorphism and male infertility in Italy. J Endocrinol Invest. 2003;26:620–2.PubMedGoogle Scholar
  29. 29.
    World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. New York: Cambridge University Press; 1999.Google Scholar
  30. 30.
    Wu W, Shen O, Qin Y, Lu J, Niu X, Zhou Z, et al. Methylenetetrahydrofolate reductase C677T polymorphism and the risk of male infertility: a meta-analysis. Int J Androl. 2012;35:18–24.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Cristina Camprubí
    • 1
  • Marta Pladevall
    • 1
  • Mark Grossmann
    • 2
  • Nicolás Garrido
    • 3
  • Maria C. Pons
    • 2
  • Joan Blanco
    • 1
  1. 1.Unitat de Biologia Cellular, Facultat de BiociènciesUniversitat Autònoma de BarcelonaCerdanyola del VallèsSpain
  2. 2.Unidad de Reproducción Asistida de Centro Médico TeknonBarcelonaSpain
  3. 3.Laboratorio de Andrología y Banco de SemenInstituto Universitario IVI ValenciaValenciaSpain

Personalised recommendations