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Quantitative PCR technique for the identification of microrearrangements of the AZFc region

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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose: The AZFc region spans about 3.5 Mb and contains many amplicons causing recombination events. Several papers have reported the occurrence of AZFc partial deletions resulting from non allelic homologous recombination (NAHR) (“gr-gr”, “b1-b3” or “b2-b3” deletions), particularly in infertile patients. DAZ genes are present in 4 copies and rearrangements involve a modification of the number of DAZ genes.

Methods: In addition to STS plus/minus PCR, we developed a quantitative technique using real time PCR (Q-PCR) to determine the number of DAZ genes. Fourteen DNA controls were selected to validate the use of Q-PCR to detect AZFc microrearrangements, and sperm DNA samples from 30 fertile men were studied.

Results: Rearrangements of 14 controls were well identified with Q-PCR, and 2 AZFc partial deletions were detected in fertile men (1 “gr-gr” and 1 “b2-b3”).

Conclusion: Q-PCR represents a well-adapted method to detect microrearrangements of the Y-chromosome, complementary to STS analysis.

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References

  1. Kuroda-Kawaguchi T, Skaletsky H, Brown LG, Minx PJ, Cordum HS, Waterston RH, Wilson RK, Silber S, Oates R, Rozen S, Page DC. The AZFc region of the Y chromosome features massive palindromes and uniform recurrent deletions in infertile men. Nat Genet 2001;29:279–86

    Article  PubMed  CAS  Google Scholar 

  2. Skaletsky H, Kuroda-Kawaguchi T, Minx PJ, Cordum HS, Hillier L, Brown LG, Repping S, Pyntikova T, Ali J, Bieri T, Chinwalla A, Delehaunty A, Delehaunty K, Du H, Fewell G, Fulton L, Fulton R, Graves T, Hou SF, Latrielle P, Leonard S, Mardis E, Maupin R, McPherson J, Miner T, Nash W, Nguyen C, Ozersky P, Pepin K, Rock S, Rohlfing T, Scott K, Schultz B, Strong C, Tin-Wollam A, Yang SP, Waterston RH, Wilson RK, Rozen S, Page DC. The male-specific region of the human Y chromosome is a mosaic of discrete sequence classes. Nature 2003;423:825–37

    Article  PubMed  CAS  Google Scholar 

  3. Repping S, Skaletsky H, Brown L, Van Daalen SKM, Korver CM, Pyntikova T, Kuroda-Kawaguchi T, de Vries JWA, Oates RD, Silber S, Van Der Veen F, Page DC, Rozen S. Polymorphism for a 1.6 Mb deletion of the human Y chromosome persists through balance between recurrent mutation and haploid selection. Nat Genet 2003;35:247–51

    Article  PubMed  CAS  Google Scholar 

  4. Repping S, Skaletsky H, Lange J, Silber S, Van Der Veen F, Oates RD, Page DC, Rozen S. Recombination between palindromes P5 and P1 on the human Y chromosome causes massive deletions and spermatogenic failure. Am J Hum Genet 2002;71:906–22

    Article  PubMed  Google Scholar 

  5. Repping S, van Daalen SK, Korver CM, Brown LG, Marszalek JD, Gianotten J, Oates RD, Silber S, Van Der Veen F, Page DC, Rozen S. 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;83:1046–52

    Article  PubMed  CAS  Google Scholar 

  6. Foresta C, Moro E, Garolla A, Onisto M, Ferlin A. Y chromosome microdeletions in cryptorchidism and idiopathic infertility. J Clin Endoc Metab 1999;84:3660–65

    Article  CAS  Google Scholar 

  7. Vogt PH, Edelmann A, Kirsch S, Henegariu O, Hirschmann P, Kiesewetter F, Kohn FM, Schill WB, Farah S, Ramos C, Hartmann M, Hartschuh W, Meschede D, Behre HM, Castel A, Nieschlag E, Weidner W, Grone HJ, Jung A, Engel W, Haidl G. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet 1996;5:933–43

    Article  PubMed  CAS  Google Scholar 

  8. Oates RD, Silber S, Brown LG, Page DC. Clinical characterization of 42 oligospermic or azoospermic men with microdeletion of the AZFc region of the Y chromosome, and of 18 children conceived via ICSI. Hum Reprod 2002;17:2813–24

    Article  PubMed  CAS  Google Scholar 

  9. Saxena R, de Vries JWA, Repping S, Alagappan RK, Skaletsky H, Brown LG, Ma P, Chen E, Hoovers JMN, Page DC. Four DAZ genes in two clusters found in the AZFc region of the human Y chromosome. Genomics 2000;67:256–67

    Article  PubMed  CAS  Google Scholar 

  10. Fernandes S, Paracchini S, Meyer LH, Floridia G, Tyler-Smith C, Vogt PH. A large deletion removes DAZ3/DAZ4 and nearby genes from men in Y haplogroup N. Am J Hum Genet 2004;74:180–87

    Article  PubMed  CAS  Google Scholar 

  11. Machev N, Saut N, Longepied G, Terriou P, Navarro A, Levy N, Guichaoua M, Metzler-Guillemain C, Collignon P, Frances AM, Belougne J, Clemente E, Chiaroni J, Chevillard C, Durand C, Ducourneau A, Pech N, McElreavey K, Mattei MG, Mitchell MJ. Sequence family variant loss from the AZFc interval of the human Y chromosome, but not gene copy loss, is strongly associated with male infertility. J Med Genet 2004;41:814–25

    Article  PubMed  CAS  Google Scholar 

  12. 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;20:191–97

    Article  PubMed  CAS  Google Scholar 

  13. de Vries JW, Repping S, van Daalen SK, Korver CM, Leschot NJ, Van Der Veen F. Clinical relevance of partial AZFc deletions. Fertil Steril 2002;78:1209–14

    Article  PubMed  Google Scholar 

  14. Repping S, de Vries JW, Van Daalen SK, Korver CM, Leschot NJ, Van Der Veen F. The use of sperm HALO-FISH to determine DAZ gene copy number. Mol Hum Reprod 2003;9:183–88

    Article  PubMed  CAS  Google Scholar 

  15. Lin YW, Thi DAD, Kuo PL, Hsu CC, Huang BD, Yu YH, Vogt PH, Krause W, Ferlin A, Foresta C, Bienvenu T, Schempp W, Yen PH. Polymorphisms associated with the DAZ genes on the human Y chromosome. Genomics 2005;86:431–38

    Article  PubMed  CAS  Google Scholar 

  16. de Vries JW, Hoffer MJV, Repping S, Hoovers JMN, Leschot NJ, Van Der Veen F. Reduced copy number of DAZ genes in subfertile and infertile men. Fertil Steril 2002;77:68–75

    Article  PubMed  Google Scholar 

  17. Fernandes S, Huellen K, Goncalves J, Dukal H, Zeisler J, Rajpert De Meyts E, Skakkebaek NE, Habermann B, Krause W, Sousa M, Barros A, Vogt PH. High frequency of DAZ1/DAZ2 gene deletions in patients with severe oligozoospermia. Mol Hum Reprod 2002;8:286–98

    Article  PubMed  CAS  Google Scholar 

  18. Ferras C, Fernandes S, Marques CJ, Carvalho F, Alves C, Silva J, Sousa M, Barros A. AZF and DAZ gene copy-specific deletion analysis in maturation arrest and Sertoli cell-only syndrome. Mol Hum Reprod 2004;10:755–61

    Article  PubMed  CAS  Google Scholar 

  19. Morel F, Fellmann F, Roux C, Bresson J-L. Meiotic segregation analysis by FISH investigation in spermatozoa of a t(X;Y)(qter->p22::q11->qter) carrier. Cytogenet Cell Genet 2001;92:63–68

    Article  PubMed  CAS  Google Scholar 

  20. Sun C, Skaletsky H, Rozen S, Gromoll J, Nieschlag E, Oates R, Page DC. Deletion of azoospermia factor AZFa region of human Y chromosome caused by recombination between HERV15 proviruses. Hum Mol Genet 2000;9:2291–96

    Article  PubMed  CAS  Google Scholar 

  21. Rozen S, Skaletsky H, Marszalek JD, Minx PJ, Cordum HS, Waterston RH, Wilson RK, Page DC. Abundant gene conversion between arms of palindromes in human and ape Y chromosomes. Nature 2003;19;423(6942):873–76

    Article  Google Scholar 

  22. Lepretre AC, Patrat C, Mitchell M, Jouannet P, Bienvenu T. No partial DAZ deletions but frequent gene conversion events on the Y chromosome of fertile men. J Assist Reprod Genet 2005;22:141–48

    Article  PubMed  Google Scholar 

  23. Ruiz LM, Szmulewicz MN, Herrera RJ. Single-nucleotide polymorphism in repetitive sequences of the human deleted in azoospermia complex. Electrophoresis 2002;23:1021–24

    Article  PubMed  CAS  Google Scholar 

  24. Szmulewicz MN, Ruiz LM, Reategui EP, Hussini S, Herrera RJ. Single-nucleotide variant in multiple copies of a Deleted in Azoospermia (DAZ) sequence- a Human Y chromosome quantitative polymorphism. Hum Hered 2002;53:8–17

    Article  PubMed  CAS  Google Scholar 

  25. de Llanos M, Ballesca JL, Gazquez C, Margarit E, Oliva R. High frequency of gr/gr chromosome Y deletions in consecutive oligospermic ICSI candidates. Hum Reprod 2005;20(1):216–20

    Article  PubMed  CAS  Google Scholar 

  26. Giachini C, Guarducci E, Longepied G, Degl’Innocenti S, Becherini L, Forti G, Mitchell MJ, Krausz C. The gr/gr deletion(s): a new genetic test in male infertility? J Med Genet 2005;42:497–502

    Article  PubMed  CAS  Google Scholar 

  27. Lynch M, Cram DS, Reilly A, Bryan MKO, Baker HWG, de Krester DM, McLachlan RI. The Y chromosome gr/gr subdeletion is associated with male infertility. Mol Hum Reprod Advance 2005;11(7):507–12

    Article  CAS  Google Scholar 

  28. Ferlin A, Tessari A, Ganz F, Marchina E, Barlati S, Garolla A, Engl B, Foresta C. Association of partial AZFc region deletions with spermatogenic impairment and male infertility. J Med Genet 2005;42:209–13

    Article  PubMed  CAS  Google Scholar 

  29. De Palma A, Burrello N, Barone N, D'Agata R, Vicari E, Calogero AE. Patients with abnormal sperm parameters have an increased sex chromosome aneuploidy rate in peripheral leukocytes. Hum Reprod 2005;20:2153–56

    Article  PubMed  Google Scholar 

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Acknowledgemnts

The authors wish to thank Pamela Albert for proofreading the English text and Mickael J Mitchell for his collaboration in sending the DNA samples that we used as controls.

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Authors and Affiliations

  1. EA MENRT 3185 Génétique et Reproduction - IFR133 IBCT, Faculté de Médecine et de Pharmacie, Besançon, France

    Virginie Rozé

  2. Service de Génétique - Histologie – BDR and CECOS, CHU, Place St Jacques, 25030, Besançon, France

    Jean Luc Bresson

  3. Service de Génétique Médicale CHUV –1011, Lausanne, Switzerland

    Florence Fellmann

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  1. Virginie Rozé
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  2. Jean Luc Bresson
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  3. Florence Fellmann
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Correspondence to Virginie Rozé.

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Rozé, V., Bresson, J.L. & Fellmann, F. Quantitative PCR technique for the identification of microrearrangements of the AZFc region. J Assist Reprod Genet 24, 241–248 (2007). https://doi.org/10.1007/s10815-006-9055-z

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  • DOI: https://doi.org/10.1007/s10815-006-9055-z

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