RESUMO
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.
Assuntos
Anoctamina-1 , Astenozoospermia , Proteínas de Transporte de Cátions , RNA Mensageiro , Receptores Acoplados a Proteínas G , Motilidade dos Espermatozoides , Espermatozoides , Zinco , Humanos , Masculino , Astenozoospermia/metabolismo , Astenozoospermia/genética , Anoctamina-1/metabolismo , Anoctamina-1/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores Acoplados a Proteínas G/genética , Zinco/metabolismo , Espermatozoides/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Proteínas de Transporte de Cátions/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Metalotioneína/metabolismo , Metalotioneína/genética , Homeostase , Adulto , Análise do Sêmen , Relevância Clínica , Proteínas de NeoplasiasRESUMO
Sucrose, as the main product of photosynthesis, plays crucial roles in plant development. Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions. We focused on four sucrose metabolism related gene families including sucrose synthase, sucrose phosphate synthase, sucrose phosphate phosphatase and UDP-glucose pyrophosphorylase. These gene families exhibited different expansion and evolutionary history as their host genomes experienced differentiated rates of the whole genome duplication, tandem and segmental duplication, or mobile element mediated gene gain and loss. They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion. However, we have detected recent positive selection during intra-species divergence. Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses. Our studies uncovered the molecular mechanisms of gene expansion and evolution and also provided new insight into the role of positive selection in intra-species divergence. Overexpression data revealed novel biological functions of these genes in flowering time control and seed germination under normal and stress conditions.
Assuntos
Metabolismo dos Carboidratos/genética , Genes de Plantas , Família Multigênica , Plantas/genética , Plantas/metabolismo , Sacarose/metabolismo , Biomassa , Elementos de DNA Transponíveis , Evolução Molecular , Flores , Duplicação Gênica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Variação Genética , Genoma de Planta , Estudo de Associação Genômica Ampla , Genômica/métodos , Germinação/genética , Fenótipo , Filogenia , Plantas/classificação , Plantas Geneticamente Modificadas , Seleção Genética , Estresse Fisiológico/genéticaRESUMO
The Anabaena genome contains two ORFs that appear to encode glutaminases. The genes were expressed as histidine-tagged fusion proteins in Escherichia coli. The purified proteins possessed glutaminase activity using l-glutamine as the substrate, but differed in biochemical properties. All2934 showed an optimal activity at 20 degrees C and pH 6.0, with a higher affinity for l-glutamine than All4774, which had optimal activity at 37 degrees C and pH 7.5. Remarkably, the glutaminase activity of All2934 was phosphate dependent, while All4774 was phosphate independent. The expression of all2934 and all4774 was analyzed using semi-quantitative reverse transcriptase-PCR. The expression level of all2934 was much higher than that of all4774 under normal and nitrogen-depletion conditions, indicating that All2934 may play an important role in metabolizing glutamine in Anabaena.