2009 Vienna - European Society of Human Genetics

Cytogenetics
spermia, 11 oligospermia) and 30 controls were recruited. Chromosome
analysis were performed from peripheral blood and Y- chromosome
deletions were detected by using commercial Kit. Sperm DNA
purifications were carried out from both patients and controls. Mitochondrial
ND1, ATPase6 and Cytb genes were amplified and the amplicons
were analysed by direct sequencing.
All the participants were 46, XY with no Y- deletions. Some common
polymorphisms/ mutations including patients and controls were detected
in ND1, ATPase6 and Cytb genes.
No correlation was found between sperm parameters and mtDNA mutations
in this study. It is suggested that low sample number may be a
reason. We are looking forward to increase the number of cases and
determine D-loop which is another important locus of mtDNA in terms
of male infertility.
P03.193
cAG repeat length in the androgen receptor gene in infertile men
in serbia
M. L. Ristanovic1 , C. Tulic2 , J. Trifunovic1 , A. Ristanovic1 ;
1 2 Institute of Human genetics, Belgrade, Serbia, Institute of Urology and Nephrology,
Belgrade, Serbia.
The purpose of this study was to evaluate CAG repeat length in the
androgen receptor gene in Serbian men with severe oligozoospermia.
Initially, 180 infertile patients were included in the study and spermogram
has been performed in order to determine the sperm density.
Patients were excluded if clinical evidence of obstructive azoospermia,
known cytogenetic defects, Y chromosome microdeletion or abnormal
hormonal parameters were present. Control group of 100 age-matched
men who had fathered at least two children was also analyzed for CAG
repeat length in the androgen receptor gene. The screening was performed
in 106 selected patients with idiopathic infertility by polymerase
chain reaction (PCR) method on DNA extracted from peripheral blood.
The mean CAG repeat length in the androgen receptor gene in infertile
group (20.2±1.2) did not differ significantly than in control group
(21.0±0.7) (t=0, 0978, p>0.05) Conclusion: No significant correlation
was found in CAG repeat length between infertile men and controls in
Serbian population.
P03.194
sperm mitochondrial DNA mutations associated with reactive
oxygen species (ROs) and sperm DNA damage in male infertility
S. Venkatesh 1 , M. Shamsi 1 , M. Kumar 1 , R. Kumar 2 , N. P. Gupta 2 , R. K. Sharma
3 , P. Talwar 3 , S. Mittal 4 , R. Dada 1 ;
1 Laboratory for Molecular Reproduction and Genetics, Department of Anatomy,
All India Institute of Medical Sciences, New Delhi, India, 2 Department of Urology,
All India Institute of Medical Sciences, New Delhi, India, 3 ART centre, R&R
Hospital, New Delhi, India, 4 Department of Obstetrics and Gynaecology , All
India Institute of Medical Sciences, New Delhi, India.
Excess Reactive oxygen species (ROS) in the semen is believed to affect
the sperm function. However, the mechanism behind the elevated
ROS and impaired sperm parameters is not clear. The present study
was aimed to find the correlation between ROS levels, mtDNA mutations
and sperm DNA damage in semen of idiopathic infertile men.
Study included 50 idiopathic infertile men and 45 fertile controls. ROS
was measured by chemiluminescence assay. Whole sperm mtDNA
was sequenced by standard PCR-DNA sequencing method. Sperm
DNA damage was studied by COMET assay. Infertile group showed
significant difference in the sperm parameters compared to control
men. Infertile group showed significantly (p< 0.001) higher ROS levels
(157.76+78.88 cpm) /10 6 spermatozoa compared to fertile controls
(4.98+1.82 cpm) / 10 6 spermatozoa. mtDNA sequencing revealed that
66% of the infertile group harboured one or more nucleotide changes
in the mitochondrial genome compared to control men inspite some
common nucleotide changes (A750G, A4769G) in both the groups.
An average of 60% sperms showed (C+D) grade-higher DNA damage
comet than the control group, which had an average of 15% sperm
cells showing C+D grade comet. Higher ROS in the semen infertile
men compared to the controls may be due to large number of nucleotide
changes in the mtDNA. As sperm DNA integrity is important for
successful fertilization, screening mtDNA mutations in infertile men
with severe oxidative stress may help in the better management for
treatment/ART
P03.195
MTHFR 5’UtR hypermethylation in testicular biopsy of
iranian patients with nonobstructive azoospermia: the role of
epigenetics in male infertility
M. Noruzinia, N. Khazamipour, P. Fatehmanesh, M. Keyhanee;
Sarem Research Center (SARC), Tehran, Islamic Republic of Iran.
Genetic factors involved in male infertility are not yet completely
explored. MTHFR has been shown to be involved in male infertility
through the pathway of folate metabolism. However, as contradictory
results are reported regarding polymorphisms of this gene, other
mechanism of pathogenesis like as promoter hypermethylation can be
involved. In this study we explored methylatin status of 5’ UTR region
of MTHFR in male patients with nonobstructive azoospermia with patient
who had no known anomaly of spermatogenesis.
Materials and method:
DNA from peripheral blood of 50 patients and 50 controls where extracted
by salting out method. DNA from testicular biopsy samples
of 35 patients with nonobstructive azoospermia and 5 patients with
obstructive azoospemia and normal spermatogenesis were extracted
using Roche DNA extraction kit. MSP was performed using primers
which had been designed to hybridize to CpG island in 5’UTR of MTH-
FR. PCR products were electrophoresed on 8% bisacrylamide.
Results:
Blood samples in patient and control groups showed no difference in
methylation. 53% of patients with nonobstructive azoospemia showed
the presence of methylated allele compaired to 0% in control group.
This result confirms the presence of hypermethylation in testicular
sample of patients with azoospermic male infertility (P-values=0.03