Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Detection of Partial Deletions of Y-chromosome AZFc in Infertile Men Using the Multiplex Ligation-dependent Probe Amplification Assay

Authors
David J Bunyan 1
Jonathan L.A Callaway 1
Nadja Laddach 2
1 Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, England
2 MRC-Holland, Amsterdam, Netherland
Abstract
Background: In recent studies, partial deletions of the azoospermia factor c region (AZFc) on the Y-chromosome have been detected in males with infertility problems. However, there has been a lot of debate about their significance. In order to study such deletions, a simple but accurate method for their detection was applied in this study. Methods: We present data obtained from the Multiplex Ligation-dependent Probe Amplification (MLPA) assay using a new Y-chromosome-specific MLPA probemix (P360) which allows the easy detection of partial AZFc deletions. Results: Partial AZFc deletions were detected in 8% of our cohort of previously mutation-negative infertile males (and 0% of the fertile control cohort). Conclusion: These results provide further evidence of the causality of partial AZFc deletions. None of the partial AZFc deletions were detectable by the standard multiplex PCR method, demonstrating the advantage of the MLPA method.
Keywords
causality
Gene dosage
Infertility
Microdeletions
Molecular Genetics
Y
chromosome
  1. Tiepolo L, Zuffardi O. Localization of factors controlling spermatogenesis in the nonfluorescent portion of the human Y chromosome long arm. Hum Genet. 1976;34(2):119-24.
  2. Navarro-Costa P, Gonçalves J, Plancha CE. The AZFc region of the Y chromosome: at the cross-roads between genetic diversity and male infertility. Hum Reprod Update. 2010;16(5):525-42.
  3. Fernandes S, Huellen K, Goncalves J, Dukal H, Zeisler J, Rajpert De Meyts E, et al. High frequency of DAZ1/DAZ2 gene deletions in patients with severe oligozoospermia. Mol Hum Reprod. 2002;8 (3):286-98.
  4. Hucklenbroich K, Gromoll J, Heinrich M, Hohoff C, Nieschlag E, Simoni M. Partial deletions in the AZFc region of the Y chromosome occur in men with impaired as well as normal spermatogenesis. Hum Reprod. 2005;20(1):191-7.
  5. Zhang F, Li Z, Wen B, Jiang J, Shao M, Zhao Y, et al. A frequent partial AZFc deletion does not render an increased risk of spermatogenic impairment in East Asians. Ann Hum Genet. 2006;70(Pt 3):304-13.
  6. Giachini C, Guarducci E, Longepied G, Degl'Innocenti S, Becherini L, Forti G, et al. The gr/gr deletion(s): a new genetic test in male infertility? J Med Genet. 2005;42(6):497-502.
  7. Krausz C, Giachini C, Xue Y, O'Bryan MK, Gromoll J, Rajpert-de Meyts E, et al. Phenotypic variation within European carriers of the Y-chromosomal gr/gr deletion is independent of Y-chromosomal background. J Med Genet. 2009;46(1):21-31.
  8. Simoni M, Bakker E, Krausz C. EAA/EMQN best practice guidelines for molecular diagnosis of y-chromosomal microdeletions. State of the art 2004. Int J Androl. 2004;27(4):240-9.
  9. Schouten JP, McElgunn CJ, Waaijer R, Zwijnen-burg D, Diepvens F, Pals G. Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res. 2002;30(12):e57.
  10. Noordam MJ, Westerveld GH, Hovingh SE, van Daalen SK, Korver CM, van der Veen F, et al. Gene copy number reduction in the azoospermia factor c (AZFc) region and its effect on total motile sperm count. Hum Mol Genet. 2011;20(12):2457-63.
  11. Wu B, Lu NX, Xia YK, Gu AH, Lu CC, Wang W, et al. A frequent Y chromosome b2/b3 subdeletion shows strong association with male infertility in Han-Chinese population. Hum Reprod. 2007;22(4): 1107-13.
  12. Yang Y, Ma M, Li L, Zhang W, Xiao C, Li S, et al. Evidence for the association of Y-chromosome haplogroups with susceptibility to spermatogenic failure in a Chinese Han population. J Med Genet. 2008;45(4):210-5.
  13. Giachini C, Laface I, Guarducci E, Balercia G, Forti G, Krausz C. Partial AZFc deletions and duplications: clinical correlates in the Italian population. Hum Genet. 2008;124(4):399-410.
  14. Repping S, Skaletsky H, Brown L, van Daalen SK, Korver CM, Pyntikova T, et al. Polymorphism for a 1.6-Mb deletion of the human Y chromosome persists through balance between recurrent mutation and haploid selection. Nat Genet. 2003;35(3): 247-51.
  15. Repping S, van Daalen SK, Korver CM, Brown L G, Marszalek JD, Gianotten J, et al. A family of human Y chromosomes has dispersed throughout northern Eurasia despite a 1.8-Mb deletion in the azoospermia factor c region. Genomics. 2004;83 (6):1046-52.
  16. Bunyan DJ, Eccles DM, Sillibourne J, Wilkins E, Thomas NS, Shea-Simonds J, et al. Dosage analysis of cancer predisposition genes by multiplex ligation-dependent probe amplification. Br J Cancer. 2004;91(6):1155-9.
  17. Archer HL, Whatley SD, Evans JC, Ravine D, Huppke P, Kerr A, et al. Gross rearrangements of the MECP2 gene are found in both classical and atypical Rett syndrome patients. J Med Genet. 2006;43(5):451-6.
  18. Benito-Sanz S, Gorbenko del Blanco D, Huber C, Thomas NS, Aza-Carmona M, Bunyan D, et al. Characterization of SHOX deletions in Leri-Weill dyschondrosteosis (LWD) reveals genetic hetero-geneity and no recombination hotspots. Am J Hum Genet. 2006;79(2):409-14.
  19. Girirajan S, Mendoza-Londono R, Vlangos CN, Dupuis L, Nowak NJ, Bunyan DJ, et al. Smith-Magenis syndrome and Moyamoya disease in a patient with del(17)(p11.2p13.1). Am J Med Genet A. 2007;143A(9):999-1008.
  20. Bakrania P, Robinson DO, Bunyan DJ, Salt A, Martin A, Crolla JA, et al. SOX2 anophthalmia syndrome: 12 new cases demonstrating broader phenotype and high frequency of large gene deletions. Br J Ophthalmol. 2007;91(11):1471-6.
  21. Blyth M, Foulds N, Turner C, Bunyan D. Severe Marfan syndrome due to FBN1 exon deletions. Am J Med Genet A. 2008;146A(10):1320-4.
  22. Wyatt A, Bakrania P, Bunyan DJ, Osborne RJ, Crolla JA, Salt A, et al. Novel heterozygous OTX2 mutations and whole gene deletions in anophthalmia, microphthalmia and coloboma. Hum Mutat. 2008;29(11):E278-83.
  23. Foley P, Bunyan D, Stratton J, Dillon M, Lynch S A. Further case of Rubinstein-Taybi syndrome due to a deletion in EP300. Am J Med Genet A. 2009; 149A(5):997-1000.
  24. Thomas NS, Harvey JF, Bunyan DJ, Rankin J, Grigelioniene G, Bruno DL, et al. Clinical and molecular characterization of duplications encompassing the human SHOX gene reveal a variable effect on stature. Am J Med Genet A. 2009;149A (7):1407-14.
  25. Bunyan DJ, Skinner AC, Ashton EJ, Sillibourne J, Brown T, Collins AL, et al. Simultaneous MLPA-based multiplex point mutation and deletion analysis of the dystrophin gene. Mol Biotechnol. 2007; 35(2):135-40.
  26. Barber JC, Maloney VK, Huang S, Bunyan DJ, Cresswell L, Kinning E, et al. 8p23.1 duplication syndrome; a novel genomic condition with unexpected complexity revealed by array CGH. Eur J Hum Genet. 2008;16(1):18-27.
  27. Glancy M, Barnicoat A, Vijeratnam R, de Souza S, Gilmore J, Huang S, et al. Transmitted duplication of 8p23.1-8p23.2 associated with speech delay, autism and learning difficulties. Eur J Hum Genet. 2009;17(1):37-43.
  28. Barber JC, Bunyan D, Curtis M, Robinson D, Morlot S, Dermitzel A, et al. 8p23.1 duplication syndrome differentiated from copy number variation of the defensin cluster at prenatal diagnosis in four new families. Mol Cytogenet. 2010;3:3.
Journal of Reproduction & Infertility
Volume 13, Issue 3
Summer 2012
Pages 174-178