Analysis of the AZF Region in Slovak Men with Azoospermia
Y chromosome microdeletions are the most common genetic cause of male infertility and screening for these microdeletions in azoospermic or severely oligospermic men is now standard practice. Analysis of the Y chromosome in men with azoospermia or severe oligozoospermia has resulted in the identification of three regions in the euchromatic part of the long arm of the human Y chromosome (Yq11) that are frequently deleted in men with otherwise unexplained spermatogenic failure. PCR analysis of microdeletions in the AZFa, AZFb and AZFc regions of the human Y chromosome is an important screening tool. The aim of this study was to analyse the type of microdeletions in men with fertility disorders in Slovakia. We evaluated 227 patients with azoospermia and with normal karyotype. All patient samples were analyzed cytogenetically. For PCR amplification of sequence-tagged sites (STS) of the AZFa, AZFb and AZFc regions of the Y chromosome was used Devyser AZF set. Fluorescently labeled primers for all markers in one multiplex PCR reaction were used and for automated visualization and identification of the STS markers we used genetic analyzer ABi 3500xl (Life Technologies). We reported 13 cases of deletions in the AZF region 5,73%. Particular types of deletions were recorded in each region AZFa,b,c .The presence of microdeletions in the AZFc region was the most frequent. The study confirmed that percentage of microdeletions in the AZF region is low in Slovak azoospermic patients, but important from a prognostic view.
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 J. Poongothai, T. S.Gopenath, S. Manonayaki "Genetics of human male inferertility.” Singapore Med,,vol.50, pp.336-347,April 2009.
 K.L.O´Flynn,O´Brien,A.C.Varghese,A.Agarwali, "The genetic causes of male factor infertility : a review.”Fertil Steril,vol.93,no.1,pp.1-12,Jan 2010.
 B.R.Emery,D.T.Carrell,”The effect of epigenetic sperm abnormalities on early embryogenesis.” Asian J,Androl.,vol.8,no21,pp.131-142,March 2006.
 G.D.Palermo,L.T.Coôombero,J.J.Harriprashad,P.N.Schlegel,Z.Rosen- Waks,”Chromosome analysis of epididymal and testicular sperm in azoospermic patients undergoing ICSI.” Hum.Reprod, vol.17, no.3,, pp.570-575,March 2002.
 R.Slezak,M.Sasiadek,”Chromosome Y microdeletions in the pathogenesis of male infertility.”Pol Merkur Lekarski,vol.13 ,no.75, pp.229-233, Sep 2002.
 C.Britom-Jones,C.J.Haines,”Microdeletions on the long arm of the Y chromosome and their association with male factor infertility.”Hong Kong Med.J.,vol.6,no.2,pp.184-189, June 2000.
 T.Shinka,Y.Nakahori,”The azoospermic factor on the &y chromosome.”Acta Paediatr Jpn.,vol.38,no.4,pp.399-404, Aug.1996.
 J.C.Venter,M.D.Adams,E.W.Myers, "The sequence of the human genome." Science, vol. 291, no.5507,pp.1304-1351, Feb 2001.
 C.Mitchell,V.R.Harley,"Biochemical defects in eight SRY missense mutations causing XY gonadal dysgenesis." Mol Genet Metab, vol.77,no.3,pp217-225, Nov 2002.
 A.H.Sinclair,P.Berta,M.Palmer,J.R.Hawkins,B.L.Griffiths,M.J.Smith,J. W.Foster,A.M.Frischauf,R.Lovell-Badge,P.N.Goodfellow, "A gene from the human sex-determining region encodes a protein with homology to conserved DNA-binding motif." Nature, vol.346,pp.240- 246,July 1990.
 J.C.Achermann,G.Ozisik,M.Ito,U.A.Orun,K.Harmanci,B.Gurakan,J.L.Ja meson, "Gonadal determination and adrenal development are regulated by the orphan nuclear receptor steroidogenic factor-1, in a dosedependent manner." J Clin.Endocrinol Metab, vol.87,pp.1829-1833, Feb 2002.
 A.Fwrlin,E.Moro,A.Rossi,B.Dallapiccola,C.Foresta,"The huma Y chromosome ´s azoospermia factorb (AZFb) region: sequence, structure, and deletion analysis in infertile men". J Med Genet, vol.40,no.1,pp.18- 24, Jan 2003.
 S.Simoni,E.Bakker,C.Krausz," EAA/EMQN best practice guidelines for molecular diagnosis of Y -chromosomal microdeletions. State of the art 2004. Int J Androl, vol.27,no.4, pp. 240-249, Aug 2004.
 C.Kamp,K.Huellen,S.Fernandes,M.Sousa,P.N.Schlegel,A.Mielnik,S.Kle iman,H.Yavetz,W.Krause,W.Kupker,R.Johannison,W.Schulze,W.Weidn er,A.Barros,P.H.Vogt," High deletion frequency of the complete AZFa sequence in men with Sertoli-cell-only syndrome."Molec Hum Reprod, vol. 7,no.10,pp.987-994, July 2001.
 P.H.Vogt," AZF deletions and Y chromosomal haplogroups : history ad updates on sequence." Hum.Reprod., vol. 11,no.4, pp. 319-336, Aug 2005.
 P.H.Vogt, C.L.Falcao, R.Hanstein,J.Zimmer, "AZF Proteins. " Int.J.Androl., vol. 31, no.4,pp. 383-394, May 2008.
 C.Foresta , E.Moro A. Ferlin ,” Y chromosome microdeletions and alterations of spermatogenesis.” Endocrine Reviews, vol.22,no.2, pp. 226-23, July 2013.
 S.L. SãoPedro, R. Fraietta, D. Spaine, C.S. Porto, M. Srougi, A.P. Cedenho and M.C.W. Avellar ,"Prevalence of Y chromosome deletions in a Brazilian population of nonobstructive azoospermic and severely oligozoospermic men.” Braz J Med Biol Res, vol.36 ,no.6,pp.787-793, June 2003.
 M.Totonchi,A.Mohseni Meybody,P.Borijan,P.Boroujeni,M.Sedighi Gilani,N.Amadani,H.Gourabi,”Clinical data for 185 infertile Iranian men with Y-chromosome microdeletion.”J.Assist.Reprod.Genet.,vol.29, no.8,pp.847-853, Aug 2012.
 Y.S.Zhang,R.I.Dai,R.X.Wang,H.G.Zhang,S.Chen,R.Z.Liu,”Analysis of Y chromosome microdeletion in 1738 infertile men from northeastern China.”Urology,vol.82,no.3,pp.584-588, Sep 2013.
 A.Mitra,R.Dada,R.Kumar,N.P.Gupta,K.Kucheria,S.K.Gupta,”Screening for Y-chromosome microdeletions in infertile Indian males : utility of simplified multiplex PCR.” Indian J Med.Res,vol.127,no.2,pp.124-132, Feb 2008.
 G.R.Dohle,D.J.Halley,J.O.Van Hemel,A.M.Van Den Douwel,M.H. Pieters,R.F.Weber,L.C.Govaerts,”Genetic risk factors in infertile men with severe oligozoospermia and azoospermia.” Hum Reprod,vol.17, no.1,pp.13-16, Jan 2002.
 P.J.Stahl,P.Masson,A.Mielnik,M.B.Marean,P.N.Schlegel,D.A.Paduch,” A decade of experience emphasizes that testing for Y microdeletions is essential in American men with azoospermia and severe oligozoospermia.” Fertil.Steril.,vol.94 no.5,,pp.1753-1756, Oct 2010.
 M.Nakashima,E.Koh,M.Namiki,A.Yoshida,”Multiplex sequence-tagged site PCR for efficient screening of microdeletions in Y chromosome in infertile males with azoospermia or severe oligozoospermia.” Arcg Androl.,vol.48,no.5,pp.351-358, Sep-Oct 2002.
 I.Rejeb,R.M´rad,F.Maazoul,M.Trabelsi,L.Ben Jemaa,M.Caabouni,F.Zhio ua,H.Chaabouni.”Y chromosome microdeletions in Tunisian infertile males.”Pathol .Biol (Paris),vol.56,no.3,pp.111-115, May 2008.
 R.Behulova,I.Varga,L.Strhakova,A.Bozikova,D.Gabrikova,I.Boronova,V .Repiska,”Incidence of microdeletions in the AZF region of the Y chromosome in Slovak patients with azoospermia.”Biomed.Pap.Med.Fac. Univ.Palacky Olomouc,Czech Repub.,vol.155,no.1,pp.33-38, March 2011
 D.Chellat,M.L.Rezgoune,K.Mc.Elreavey,N.Kherouatou,S.Ben-Bouhadja, H.Douadi,B.Cherifa,N.Abadi,D.Satta,” First study of microdeletions in the Y chromosome of Algerian infertile men with idiopathic oligo or azoospermia.Urol .Int.,vol.90,no.4,pp.455-459, March 2013.
 M.Balkan,S.Tekes,A.Gedik,”Cytogenetic and Y chromosome microdeletion screening studies in infertile males with Oligozoospermia and Azoospermia in Southeast Turkey.”J.Assist.Reprod.Genet., vol.25,no.11-12,pp.559-565,Nov 2008.
 A.Ferlin,E.Motto,A.Garolla,C.Foresta,”Human male infertility and Y chromosome deletions: role of the YZF candidate genes DAZ,RBM and DFFRY.” Hum Reprod,vol.14,no.7,pp.1710-1716, July 1999.
 S.E.Kleiman,L.Yogev,R.Gamzu,R.Hauser,A.Botcha,G.Paz,J.B.Lessing,, Y.Yaron,H.Yavetz,”Three generations evaluation of Y-chromosome microdeletion.”J .Androl,vol20,no.3,394-398, May-June 1999.
 O.F.Khabour,A.S.Fararjeh,A.A.Alfaduri,”Genetic screening for AZF Y microdeletions in Jordan azoospermic infertile men.”Intl J Mol Epidemiol Genet.,vol.17,no.5,pp.47-50, Feb 2014.
 R. Behulova, I.Varga, L. Strhakova, A.Bozikova, D. Gabrikova,I. Boronova, V. Repiska, "Incidence of microdeletions in the AZF region of the Y chromosome I Slovak patients with azoospermia..” Biomed Pap Med Fac Univ Palacky Olomouc, Czech Repub.,vol. 155,no.1,pp.33–38, March 2011.