Polymorphic noncoding region of GNRH1 gene in male patients with impaired fertility

Abstract

Impaired male fertility is a multifactorial pathological condition and genetic contribution is up to 20%. Gonadotropin Releasing Hormone 1 (GNRH1) is associated with maturation of spermatozoa. We aimed to study genetic sequence of GNRH1 gene in azoospermia males. We collected 50 clinically diagnosed infertile males after the informed consent. DNA was extracted and primers were designed to amplify the exonic and exon/intron boundaries of exon 1 and exon 2 region of GNRH1 gene. The amplicons were sequenced and analyzed using bioninformatic tools. We have identified sequence variations at c.1325-131del.A, c.1325+40 T>G, c.1325+34T>G, c.1325+124 Ins. A, c.1325+126 Ins. A, c.1325+181T>G, c.1325+264 Ins.G and c.1325+269 Ins. A positions. Human Splicing Finder predicted that c.1325-131del.A, c.1325+34T>G, c.1325+124 Ins. A, c.1325+264 Ins.G and c.1325+269 Ins. A have impact on the splicing while c.1325+40 T>G, c.1325+126 Ins. A and c.1325+181T>G have relation with the splicing. The c.1325+40 reported in literature as rs2709608 but has no association with the male infertility. The c.1325+181T>G We could not find other variations in the literature therefore suggested to be novel. Our study suggests that intronic region of GNRH1 gene a highly variable in Pakistani infertile male patients and suggests to explore its correlation with the disease on a large scale.