Different Types Of Spermatogenesis Disorder Genome-wide Methylation Differences Analysis With Important Functional Gene Expression Validation

Objective: Application of high-density DNA methylation chip to analyze the methylation level in different types of spermatogenesis failureMethods: Collected72normal fertile male semens and different types of infertile male semen samples to do the extraction of sperm DNA and bisulfite conversion. Application of Illumina HD450K Infinium methylation BeadChip to analyze different gene methylation between different types of sperm DNA damage and normal fertile man.Results:678genes showed significant differences in the degree of methylation between the oligospermia group and normal fertile group (Diff Score=50, P<0.00001), which included568hyper-methylation genes and119hypo-methylation genes.2231genes showed significant differences in the degree of methylation between the asthenozoospermia group and normal fertile group (Diff Score=50, P<0.00001), which included1181hyper-methylation genes and420hypo-methylation genes.1550genes showed significant differences in the degree of methylation between the unexplained infertile group and normal fertile group (Diff Score=50, P<0.00001), which included1154hyper-methylation genes and396hypo-methylation genes. Cluster analysis of the candidate genes showed that:abnormal methylation genes in oligospermia men mainly participated in the apoptosis, cell cycle proteins, while abnormal methylation genes in asthenozoospermia men mainly involved in ion channels and transport protein, cytoskeleton, flagellum moveability. abnormal methylation genes in unexplained infertile men are mainly focused on allograft rejection, immune related, endocrine and metabolism disorder. These genes may be one of the reasons that leaded to different types of spermatogenesis failure.Conclusion:Based on the analyse by high-density genome-wide methylation chips,we screened out a batch of abnormal methylation genes related to the process of spermatogenesis. This study lays the foundation for further exploration of the molecular mechanism deep into different types of spermatogenesis failure. Objective:To study sperm mRNA transcript expression of MMEL1and TEKT5gene in different types of spermatogenesis failure cases.Methods:Using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) method to detect MMEL1and TEKT5gene mRNA levels in normal control group, asthenozoospermia and oligozoospermia group respectively.Results:Asthenozoospermia and oligozoospermia MMEL1and TEKT5mRNA expression levels were lower than that of normal control group. The relative expression amount of MMEL1in the normal control group was significantly higher than the asthenozoospermia group (0.696vs.0.329, P=0.000<0.05), while less significant differences compared in the oligozoospermia group (0.696vs.0.665, P=0.972>0.05). The asthenozoospermia group of MMEL1relative expression level was significantly lower than the oligozoospermia group (0.329vs.0.665, P=0.000<0.05). Statistical analysis of different samples TEKT5relative expression showed that TEKT5expression of the normal group and oligozoospermia group were significantly higher than that in the asthenozoospermia group (0.681vs.0.316, P=0.0000.05,0.527vs.0.316, P=0.008<0.05), while no significant difference between normal group and the oligozoospermia group (0.681vs.0.527, P=0.204>0.05).Conclusions:Abnormal methylation of MMEL1and TEKT5gene in different types of spermatogenesis failure may show the silencing effect. Genomic methylation plays an important role in the pathogenesis of spermatogenesis. It provides a preliminary basis for further study to elucidate the molecular mechanism of oligozoospermia and asthenozoospermia.