Download Novel sequence variations in exon 25 of JHDM2A among

P310
Novel sequence variations in exon 25 of JHDM2A among
overweight and obese sub/infertile males in South India
Mohsen Najafi and Suttur S. Malini
University of Mysore, Mysore, India
Objectives: To analyze three exons of JHDM2A gene in obese
males associated with sub/infertility and to compare with non-obese
infertile and fertile males as controls in Karnataka, South India.
Materials and methods: For the candidate gene analysis, 200 cases
with obesity/ and infertility and 50 controls (non-obese fertile males)
were selected. After 3-5 days of ejaculatory abstinence the semen
samples were collected in a sterile plastic container by the process
of masturbation from the subjects (WHO, 1999). The collected
samples were allowed to liquefy at 37°C for 30 minutes and analyzed
for semen volume, sperm count, motility, viability etc. within one
hour after collection. The blood samples of the cases and controls
were analyzed following the genomic DNA extraction and RT-PCR
amplification. The qPCR products then immediately were subjected
to High Resolution Melting analysis in order to find out the variation
in the mentioned exons. The suspected samples with variation were
sequenced for confirmation, using Applied Biosystem 3130 genetic
analyzer, and the sequences were subjected for multiples nucleotide
sequence alignment using CLC Main Workbench 6.0to analyze the
changes in nucleotide as well as amino acid sequences.
Results: Variations including single or double nucleotide insertion
as well as substitution was found in exon 25 of the candidate gene
in 11 subjects. No variation was observed in either exon 23 and 24.
Semen parameters were found to be more impaired in obese infertile
males compared to other groups in the study.
Conclusion: obesity was found to be linked with male fertility
problems. Although the JHDM2A variants were found in obese males
to be associated with subfertility or infertility, this study encourages
further comprehensive multidisciplinary research taking diverse large
human populations to understand the relationship between obesity
and male infertility.