[Expressions of zinc homeostasis proteins, GPR39 and ANO1 mRNA in the sperm of asthenozoospermia patients and their clinical significance].

OBJECTIVE: To explore the expressions of zinc homeostasis-related proteins, G protein-coupled receptor 39 (GPR39) and ANO1 mRNA in the sperm of patients with asthenozoospermia (AS), and analyze their correlation with sperm motility. METHODS: We collected semen samples from 82 male subjects with PR+NP < 40%, PR < 32% and sperm concentration > 15×106/ml (the AS group, n = 40) or PR+NP ≥ 40%, PR ≥ 32% and sperm concentration > 15×106/ml (the normal control group, n = 42). We analyzed the routine semen parameters and measured the zinc content in the seminal plasma using the computer-assisted sperm analysis system, detected the expressions of zinc transporters (ZIP13, ZIP8 and ZNT10), metallothioneins (MT1G, MT1 and MTF), GPR39, and calcium-dependent chloride channel protein (ANO1) in the sperm by real-time quantitative PCR (RT qPCR), examined free zinc distribution in the sperm by laser confocal microscopy, and determined the expressions of GPR39 and MT1 proteins in the sperm by immunofluorescence staining, followed by Spearman rank correlation analysis of their correlation with semen parameters. RESULTS: There was no statistically significant difference in the zinc concentration in the seminal plasma between the AS and normal control groups (P>0.05). Compared with the controls, the AS patients showed a significantly reduced free zinc level (P<0.05), relative expressions of MT1G, MTF, ZIP13, GPR39 and ANO1 mRNA (P<0.05), and that of the GPR39 protein in the AS group (P<0.05). No statistically significant differences were observed in the relative expression levels of ZIP8, ZNT10 and MT1 mRNA between the two groups (P>0.05). The relative expression levels of GPR39, ANO1, MT1G and MTF mRNA were positively correlated with sperm motility and the percentage of progressively motile sperm (P<0.05). CONCLUSION: The expressions of zinc homeostasis proteins (MT1G, MTF and ZIP13), GPR39 and ANO1 mRNA are downregulated in the sperm of asthenozoospermia patients, and positively correlated with sperm motility.

Association of SNPs in zinc transporter genes on seminal plasma zinc levels in humans.

This study is to examine the effects of single nucleotide polymorphisms (SNPs) of SLC30A and SLC39A on seminal plasma zinc concentration. Blood and seminal plasma samples were collected from outpatients. SNPs of zinc transporters were analyzed by next Generation sequencing technology, and seminal plasma zinc concentration were determined by inductively coupled plasma optical emission spectrometry. Our date showed nine SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, SLC39A14 rs1051708) were significantly associated with seminal plasma zinc concentration, and 14 SNPs (SLC30A8 rs2466295, rs2466294, SLC30A10 c.-160 C>G, SLC39A6 rs148550301, SLC39A8 rs9331, rs9705, rs151392, rs151393, SLC39A11 rs9912126, rs61736066, rs36041371 and SLC39A14 rs1051708, rs76963096, rs17060854) were found to be significantly associated with total zinc per ejaculate. The seminal plasma zinc concentrations and total zinc per ejaculate were associated with the number of SNPs, and decreased significantly when five SNPs (SLC39A8 rs9331, rs9705, rs151392, rs151393 and SLC39A14 rs1051708) were a combination of homozygous genotype. Our findings suggest that different zinc transporter SNPs may significantly affect seminal plasma zinc levels.