Reproduction Abstracts

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 min and analyzed for semen volume, sperm count, motility, viability, etc. within 1 h 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 6.0 to 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.