2008 Barcelona - European Society of Human Genetics

Cytogenetics
and 90 normozoospermic men . We screened simultaneously for 254
different CFTR mutations and variations using arrayed primer extension
(APEX) genotyping microarray (Asper Biotech Ltd) .
Results: CFTR mutations and variants were demonstrated in 22
(17 .7%) of 124 oligozoospermic patients and in 13 (14 .4%) of 90 control
men . In addition, the total frequency of mutant/variant alleles in
infertility group was slightly, but not significantly higher than in controls
(9 .7 vs 7 .2%) . Similar trend was also observed for IVS8-5T allele
frequencies (3 .6 vs 2 .2%, respectively) . Although we demonstrate
comparable CFTR mutation/variation frequencies in both groups, the
causal relationships between specific CFTR mutations and male infertility
cannot be completely ruled out .
P02.238
simultaneous inactivation of the transcription factors sox9 and
sox8 in murine testis development leads to complete infertility
F. J. Barrionuevo 1 , H. Scherthan 2 , C. Lécureil 3 , F. Guillou 3 , M. Wegner 4 , G.
Scherer 1 ;
1 Institute of Human Genetics, University of Freiburg, Freiburg, Germany, 2 Institut
für Radiobiologie der Bundeswehr, München, Germany, 3 Institut National de
la Recherche Agronomique, Université de Tours, Nouzilly, France, 4 Institute of
Biochemistry, University of Erlangen, Erlangen, Germany.
Heterozygous loss-of-function mutations of the HMG-box transcription
factor SOX9 result in campomelic dysplasia, a human skeletal malformation
syndrome associated with XY sex reversal . The murine Sox9
gene is expressed in embryonic and postnatal Sertoli cells of the mouse
testis, and inactivation of Sox9 before the sex determination stage at
E11 .5 leads to complete XY sex reversal . To see whether Sox9 is required
for testis development after testis induction, we crossed Sox-
9 flox mice with an AMH(Anti-Müllerian Hormone)-Cre transgenic line .
Conditional Sox9 null mutants, SOX9-negative at E14 .0, are initially
fertile, but become sterile from complete meiotic arrest at around 5
months . As Sox8, a Sox9 related transcription factor, i) is expressed
similar to Sox9 during murine testis development, ii) has been shown
in vitro to activate AMH, a Sox9 target during testis development, iii) is
expressed normally in AMH-Cre;Sox9 flox/flox mutants, and iv) as homozygous
Sox8 null mutants show no obvious early gonadal phenotype,
we hypothesized that Sox8 may compensate for the absence of Sox9 .
We therefore generated a Sox9 conditional knockout on a Sox8 mutant
background . In double mutants heterozygous for Sox8, testes develop
normally up to post-natal day 10 (P10), but subsequently show
spermatogenic arrest . Homozygous double mutants show normal testis
cord formation at E15 .5, but subsequent testis cord development
is impaired; at P6, testis cords are completely irregular in shape and
appear fibrotic, resulting in complete infertility. In summary, concerted
function of Sox9 and Sox8 in post E14 .0 Sertoli cells is essential for the
maintencance of testicular function .
P02.239
Real -time PcR evaluation of tsPY copy number in infertile and
seminoma patients.
R. Vodicka 1 , K. Krizova 1 , R. Vrtel 1 , L. Dusek 2 , J. Santavy 1 ;
1 University Hospital and Palacky University Olomouc, Olomouc, Czech Republic,
2 Institute of Biostatistik and Analyses, Masaryk University Brno, Brno, Czech
Republic.
Introduction: Multicopy TSPY gene is localized on chromosome Y in
MSY region in gene clusters . Total number of the gene copies is estimated
from 20 to 40. Testis specific expression indicates a role in
meiotic or post meiotic processes during spermatogenesis .
The aims of this work are:
1) Confirmation of our previous findings (Vodicka R, et al., TSPY gene
copy number as a potential new risk factor for male infertility . Reprod
Biomed Online . 2007 May;14(5):579-87 .)
2) Analyses and comparison of a new DNA samples from infertile and
seminoma patients .
Material and method: There were included 104 infertile and 6 seminoma
patients and 50 healthy controls into study .
Copy number relative quantification was measured using the combination
of two Real -Time PCRs by Y quantifiler kit and by SYBR green
kit .
Results: Our results confirmed increasing copy number in infertile patients
(in average 53 relative TSPY copies) compare to controls (in
average 31 relative copies) even after enlarged collection of samples .
In addition the seminoma patients showed twice as many copy number
compare to the infertile patients (in average 102 relative copies) .
Conclusion: The main importance of our findings lies in great diagnostic
potential in male infertility genetic background testing . Number
of TSPY copies could be also significant tumor marker in relation to
testicular tumorgenesis .
P02.240
AZF microdeletions on the Y chromosome of Iranian infertile
men with non-obstructive azoospermia
R. Mirfakhraie 1,2 , S. M. Kalantar 3 , M. Montazeri 2 , N. Salsabili 4 , G. Modabber 2 ,
S. M. Seyed Hassani 3 , M. Houshmand 2 , F. Mirzajani 2 ;
1 Islamic Azad University of Tehran, Science & Research Campus, Tehran, Islamic
Republic of Iran, 2 National Institute for Genetic Engineering & Biotechnology,
Tehran, Islamic Republic of Iran, 3 Research & Clinical Centre for Infertility,
Yazd, Islamic Republic of Iran, 4 Mirza Kouchak Khan Hospital,Tehran University
of Medical Sciences, Tehran, Islamic Republic of Iran.
The human Y chromosome contains genes that are essential for spermatogenesis
specially those that are located on three major intervals
defined as AZFa, AZFb and AZFc.
Deletions in these genes may result in spermatogenic failure in patients
with non-obstructive azoospermia and oligozoospermia . Widely
different frequencies of such deletions (0-55%) have been reported
from different populations .
The main purpose of this study is to detect the frequency of Y chromosome
microdeletions in Iranian patients with non-obstructive azoospermia
and fertile control subjects . Multiplex polymerase chain reaction
(PCR) was applied using several sequence-tagged site (STS)
primer sets, in order to determine Y chromosome microdeletions in
100 infertile males and 50 fertile controls .
Microdeletions in AZFa, AZFb and AZFc (DAZ gene) regions were only
detected in seventeen of the patients (17%) with the frequency of 15%,
49%, and 36% respectively .
Our findings suggest that knowing the prevalence of AZF microdeletions
in Iranian infertile men will be informative before starting assisted
reproductive treatments .
P02.241
male infertility and biotransformation enzyme gene
polymorphisms
M. Volk 1 , A. Kastrin 1 , H. Jaklič 1 , B. Zorn 2 , B. Peterlin 1 ;
1 Institute of Medical Genetics, University Medical Center Ljubljana, Ljubljana,
Slovenia, 2 Department of Andrology, University Medical Center Ljubljana, Ljubljana,
Slovenia.
Environmental xenobiotics such as organophosphate pesticides are
known to be involved in male infertility . Interindividual genetic variations
in biotransformation enzyme activities can lead to differences
in the susceptibility to male infertility . In this case-control study, PCR
was used to investigate the association between polymorphisms in
the PON and GST genes (PON1-55/192, PON2-311, GSTM1/T1) and
male infertility in 381 Slovenian participants (the study group of 187
infertile male participants: 86 with azoospermia, 101 with oligoasthenoteratozoospermia;
the control group of 194 fertile males) .
We found statistical significant difference in the PON1-55 genotype
distribution between the infertile and fertile men (chi-square(2) = 7 .37;
p = 0 .02), which after applying Bonferroni correction was no longer
significant. Likewise, no significant differences in frequencies of genotypes
of other tested polymorphisms, PON1-192, PON2-311, GSTM1/
T1, respectively, and the occurrence of male infertility were observed
(Table 1) .
In this case-control study we didn’t confirm the association between
PON1/2 or GSTM1/T1 genetic variations and male infertility in Slovenian
participants . However, limitations of the genetic association studies,
namely, the relatively small sample size and population specific
genotype effects which make results difficult to reproduce, should be
considered when interpreting and generalizing the results .
Table 1 . Genotype frequencies of the PON and GST polymorphisms in
infertile and control group .