RESUMEN
Hyperpolarization is associated with decreased intracellular Na+ concentration through the closure of the epithelial Na+ channels (ENaCs) during capacitation. 5'-AMP-activated protein kinase (AMPK) is involved in the regulation of Na+ transport by reducing ENaC-ß abundance in the plasma membrane in somatic cells. However, it is not known whether AMPK acts on ENaCs in sperm. The aim of the present study was to analyze the role of AMPK activation in the regulation of ENaC and to examine its relationship with capacitation-associated hyperpolarization of human sperm. Human sperm were treated with AICAR (AMPK activator) in non-capacitating and capacitating conditions. AMPK activity and ENaC-ß concentration were evaluated by ELISA. Flow cytometry was used to measure tyrosine phosphorylation, hyperpolarization, intracellular Na+ concentration and acrosome reaction. Immunofluorescence staining was carried out to analyze the distribution of ENaC-ß and CD46 in sperm. We found that induction of capacitation triggered AMPK phosphorylation. AMPK activation by AICAR increased tyrosine phosphorylation. AICAR decreased ENaC-ß levels, mainly localized at the principal-piece of the flagellum, resulting in lower intracellular Na+ concentration and increased hyperpolarization of the plasma membrane. Altogether, these data provide evidence that AMPK activation is involved in capacitation-associated hyperpolarization by reducing ENaC abundance in human sperm.
Asunto(s)
Canales Epiteliales de Sodio , Capacitación Espermática , Proteínas Quinasas Activadas por AMP/metabolismo , Canales Epiteliales de Sodio/metabolismo , Humanos , Masculino , Fosforilación , Semen/metabolismo , Sodio/metabolismo , Espermatozoides , Tirosina/metabolismoRESUMEN
This study aimed to investigate the difference between infertile men and healthy (normozoospermic) men in terms of demographic characteristics, dietary habits, anthropometric measurements, and body composition. We included 80 males (40 subfertile and 40 healthy normozoospermic) between the ages of 25 and 54 years. Information was obtained from the participants regarding their socio-demographic characteristics, health status, dietary habits, and food intake. Food frequency questionnaires, food records, anthropometric measurements, body composition, and sperm analysis were statistically evaluated using IBM SPSS Statistics 20 programme. The findings of this study showed that the mean BMI of the subfertile group was significantly higher than that of the normozoospermic group. The frequency of eating out was significantly higher in the subfertile group than in the normozoospermic group. It was also determined that the consumption of fish was significantly lower; in contrast, consumption of sugar sweetened beverages, and alcohol was significantly higher in the subfertile group than in the normozoospermic group. Moreover, it was found that sugar sweetened bevareges, red meat, organ meats consumption are negatively; and that fish, egg, nut consumption are positively correlated with sperm parameters. In summary, in men receiving infertility treatment, excessive consumption of meat and sugary drinks should be considered cautiously. However, fish, nuts and eggs consumption should be provided in line with the nutrition guidelines.
Asunto(s)
Dieta/efectos adversos , Sacarosa en la Dieta/efectos adversos , Conducta Alimentaria , Fertilidad , Infertilidad Masculina , Adulto , Sacarosa en la Dieta/administración & dosificación , Ingestión de Alimentos , Humanos , Masculino , Persona de Mediana Edad , Encuestas Nutricionales , Salud Reproductiva , Análisis de Semen , Encuestas y Cuestionarios , TurquíaRESUMEN
PURPOSE: To compare the effect of two different sperm preparation techniques, including swim-up and gradient methods on sperm deoxyribonucleic acid (DNA) fragmentation status of semen samples from unexplained and mild male factor subfertile patients undergoing intrauterine insemination (IUI). DESIGN: A prospective randomized study was conducted in 65 subfertile patients, including 34 unexplained and 31 male factor infertility to compare basal and post-procedure DNA fragmentation rates in swim-up and gradient techniques. Sperm DNA fragmentation rates were evaluated by a sperm chromatin dispersion (SCD) test in two portions of each sample of semen that was prepared with either swim-up or gradient techniques. Sperm motility and morphology were also assessed based on WHO 2010 criteria. RESULTS: Swim-up but not gradient method yielded a statistically significant reduction in the DNA fragmented sperm rate after preparation as compared to basal rates, in the semen samples of both unexplained (41.85 ± 22.04 vs. 28.58 ± 21.93, p < 0.001 for swim-up; and 41.85 ± 22.04 vs. 38.79 ± 22.30, p = 0.160 for gradient) and mild male factor (46.61 ± 19.38 vs. 30.32 ± 18.20, p < 0.001 for swim-up and 46.61 ± 19.38 vs. 44.03 ± 20.87, p = 0.470 for gradient) subgroups. CONCLUSIONS: Swim-up method significantly reduces sperm DNA fragmentation rates and may have some prognostic value on intrauterine insemination in patients with decreased sperm DNA integrity.
Asunto(s)
Centrifugación por Gradiente de Densidad/métodos , Fragmentación del ADN , Infertilidad Masculina , Inseminación Artificial/métodos , Manejo de Especímenes/métodos , Motilidad Espermática/fisiología , Adulto , Femenino , Humanos , Masculino , Estudios ProspectivosRESUMEN
OBJECTIVE: This study aimed to evaluate the effect of luteal phase support on clinical pregnancy and live birth rates after ovulation induction and intrauterine insemination (IUI). METHODS: 579 cycles from 2010 to 2013 were retrospectively evaluated. Ovarian stimulation was performed with gonadotropins, and rHCG was used for ovulation triggering. All patients received IUI. 451 cycles were supported by receiving vaginal micronized progesterone capsules (142 cycles) or vaginal progesterone gel (309 cycles) whereas 128 cycles were not supported. RESULTS: Clinical pregnancy (20.6 versus 9.4%; p = 0.004) and live birth rates (14 versus 7%; p = 0.036) were higher for supported group than for unsupported group. Progesterone gel and micronized progesterone subgroups achieved similar clinical pregnancy and live birth rates (21.4 versus 19%, p = 0.567 and 14.2 versus 13.4%, p = 0.807; respectively). CONCLUSIONS: Luteal phase support improved the success of IUI cycles affecting both clinical pregnancy and live birth rates when gonadotropins were used for ovulation induction. The use of vaginal progesterone gel or micronized progesterone significantly improves clinical pregnancy rates. The live birth rates were higher in the progesterone gel group, but were similar in the micronized progesterone group compared to the unsupported group.
Asunto(s)
Mantenimiento del Cuerpo Lúteo/efectos de los fármacos , Fertilización In Vitro/métodos , Inseminación Artificial , Fase Luteínica/efectos de los fármacos , Inducción de la Ovulación/métodos , Progesterona/uso terapéutico , Administración Intravaginal , Adulto , Tasa de Natalidad , Femenino , Humanos , Embarazo , Índice de Embarazo , Progesterona/administración & dosificación , Estudios Retrospectivos , Adulto JovenRESUMEN
PURPOSE: To compare the efficacy of gradient and swim-up semen preparation techniques on pregnancy rates in couples undergoing intrauterine insemination (IUI) cycles with low dose gonadotropin stimulation with the diagnosis of unexplained or mild male subfertility. METHODS: Two hundred and twenty three couples were randomized into swim up or gradient technique groups for sperm preperation. The clinical and on going pregnancy rates per cycle and per patient were evaluated. RESULTS: Both clinical and ongoing pregnancy rates per cycle were significantly higher in the "gradient" group (19% and 16.9%) in comparision with the "swim up" group (9.7% and 6.9%) (p < 0.05). Clinical pregnancy and on-going pregnancy rates per patient were higher in the "gradient" group (26.1% and 23.4%) when compared to the "swim up" group (15.2% and 10.7%), (p < 0.05). In the subgroup of 191 unexplained subfertile couples with 290 cycles; the "gradient" group also revealed significantly higher clinical and ongoing pregnancy rates per cycle (21.6% and 17.9%) when compared with the "swim up" group (10.3% and 7.1%) (p < 0.05). In total of 48 treatment cycles upon 32 couples with mild male factor subfertility no significant difference were found between the two sperm preparation techniques in terms of clinical (% 5.3 vs %6.9, p > 0.05) and ongoing (% 5.3 vs %6.9, p > 0.05) pregnancy rates per cycle. CONCLUSION: The gradient technique significantly improves clinical outcome in IUI cycles of unexplained subfertile couples when compared to swim up technique. In male subfertile patients, both techniques yield similar clinical outcomes.
Asunto(s)
Inseminación Artificial Homóloga , Análisis de Semen/métodos , Adulto , Femenino , Humanos , Infertilidad Masculina/patología , Masculino , Inducción de la Ovulación , Embarazo , Índice de EmbarazoRESUMEN
Genes critical for fertility are highly conserved in mammals. Interspecies DNA sequence variation, resulting in amino acid substitutions and post-transcriptional modifications, including alternative splicing, are a result of evolution and speciation. The mammalian follicle-stimulating hormone receptor (FSHR) gene encodes distinct species-specific forms by alternative splicing. Skipping of exon 2 of the human FSHR was reported in women of North American origin and correlated with low response to ovarian stimulation with exogenous follicle-stimulating hormone (FSH). To determine whether this variant correlated with low response in women of different genetic backgrounds, we performed a blinded retrospective observational study in a Turkish cohort. Ovarian response was determined as low, intermediate or high according to retrieved oocyte numbers after classifying patients in four age groups (<35, 35-37, 38-40, >40). Cumulus cells collected from 96 women undergoing IVF/ICSI following controlled ovarian hyperstimulation revealed four alternatively spliced FSHR products in seven patients (8%): exon 2 deletion in four patients; exon 3 and exons 2 + 3 deletion in one patient each, and a retention of an intron 1 fragment in one patient. In all others (92%) splicing was intact. Alternative skipping of exons 2, 3 or 2 + 3 were exclusive to low responders and was independent of the use of agonist or antagonist. Interestingly, skipping of exon 3 occurs naturally in the ovaries of domestic cats--a good comparative model for human fertility. We tested the signaling potential of human and cat variants after transfection in HEK293 cells and FSH stimulation. None of the splicing variants initiated cAMP signaling despite high FSH doses, unlike full-length proteins. These data substantiate the occurrence of FSHR exon skipping in a subgroup of low responders and suggest that species-specific regulation of FSHR splicing plays diverse roles in mammalian ovarian function.