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.
[1] J. Poongothai, T. S.Gopenath, S. Manonayaki "Genetics of human male
inferertility.” Singapore Med,,vol.50, pp.336-347,April 2009.
[2] 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.
[3] 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.
[4] 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.
[5] 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.
[6] 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.
[7] T.Shinka,Y.Nakahori,”The azoospermic factor on the &y
chromosome.”Acta Paediatr Jpn.,vol.38,no.4,pp.399-404, Aug.1996.
[8] 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.
[9] 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.
[10] 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.
[11] 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.
[12] 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.
[13] 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.
[14] 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.
[15] 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.
[16] P.H.Vogt, C.L.Falcao, R.Hanstein,J.Zimmer, "AZF Proteins. "
Int.J.Androl., vol. 31, no.4,pp. 383-394, May 2008.
[17] C.Foresta , E.Moro A. Ferlin ,” Y chromosome microdeletions and
alterations of spermatogenesis.” Endocrine Reviews, vol.22,no.2, pp.
226-23, July 2013.
[18] 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.
[19] 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.
[20] 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.
[21] 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.
[22] 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.
[23] 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.
[24] 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.
[25] 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.
[26] 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
[27] 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.
[28] 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.
[29] 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.
[30] 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.
[31] 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.
[32] 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.
[1] J. Poongothai, T. S.Gopenath, S. Manonayaki "Genetics of human male
inferertility.” Singapore Med,,vol.50, pp.336-347,April 2009.
[2] 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.
[3] 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.
[4] 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.
[5] 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.
[6] 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.
[7] T.Shinka,Y.Nakahori,”The azoospermic factor on the &y
chromosome.”Acta Paediatr Jpn.,vol.38,no.4,pp.399-404, Aug.1996.
[8] 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.
[9] 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.
[10] 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.
[11] 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.
[12] 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.
[13] 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.
[14] 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.
[15] 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.
[16] P.H.Vogt, C.L.Falcao, R.Hanstein,J.Zimmer, "AZF Proteins. "
Int.J.Androl., vol. 31, no.4,pp. 383-394, May 2008.
[17] C.Foresta , E.Moro A. Ferlin ,” Y chromosome microdeletions and
alterations of spermatogenesis.” Endocrine Reviews, vol.22,no.2, pp.
226-23, July 2013.
[18] 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.
[19] 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.
[20] 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.
[21] 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.
[22] 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.
[23] 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.
[24] 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.
[25] 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.
[26] 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
[27] 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.
[28] 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.
[29] 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.
[30] 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.
[31] 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.
[32] 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.
@article{"International Journal of Medical, Medicine and Health Sciences:55960", author = "J. Bernasovská and R. Lohajová Behulová and E. Petrejčiková and I. Boroňová and I. Bernasovský", title = "Analysis of the AZF Region in Slovak Men with Azoospermia", abstract = "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.
", keywords = "AZF, male infertility, microdeletions, Y
chromosome.", volume = "8", number = "9", pages = "567-4", }
