Determining daz genes copy number in idiopathic non-obstructive azoospermic and oligospermic men

Alimardanian, L.; Ahani, A.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p14
Academic Journal
Introduction: Three distinct Azoospermia Factor (AZF) regions on the long arm of the Y chromosome (AZFa, AZFb, and AZFc) are essential for normal spermatogenesis. Four copies of the DAZ genes within the AZFc region are 99.99% identical which have the highest frequency of deletion in men with azoospermia and oligozoospermia. The multicopy nature of DAZ has limited the understanding of its actual role in human spermatogenesis. The current study was designed to evaluate the role of DAZ deletions and duplications in spermatogenesis. Materials and Methods: Totally, 115 azoospermic, 41 oligozoospermic and 113 fertile healthy men as controls were recruited for this study. AZFc region deletions and DAZ copy number changes were analyzed using six STS markers sY1191, sY1161, sY1197, sY1291, sY1206, sY1201. The presence and absence of each DAZ gene was evaluated using two STS markers sY581, sY587 by PCR and RFLP. Also copy numbers were analyzed by comparison of number of DAZ genes to the autosomal DAZL by Quantitative PCR followed by fragment analysis. Results: The gr/gr deletions were seen in 6.9, 7.3 and 1.7% of azoospermic, oligozoospermic and fertile men, respectively. On the other hand, b2/b3, b2/b4 deletions were found only in azoospermic group with a frequency of 4.3, 1.7% in each deleted region. And whole AZFc region deletions were found in 0.86 and 0.88% of the azoospermic and fertile men, respectively. Results of fragment analysis showed that deletions of DAZ copies with 1:1 ratio (DAZ/DAZL) were 5.2, 7.3 and 0.88% in azoospermic, oligozoospermic men and controls, respectively. DAZ copies with 0:1 ratio was 26.08 and 0.8% in azoospermic and fertile men, respectively. DAZ copies duplications with 3:1 ratio was 7.8, 2.4 and 7.9% and with 4:1 ratio was 16.5, 17.01 and 4.4% in azoospermic, oligozoospermic men and controls, respectively. Conclusion: The results of RFLP comparing to the STS-PCRs were not analyzable, suggesting a specific Y haplo groups in Iranian population that differ to the previous known haplo groups. Decreasing of DAZ copy number was associated with oligozoospermia, while neither azoospermia nor oligozoospermia was associated with increasing of DAZ copies.


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