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1.
Am J Hum Genet ; 108(2): 309-323, 2021 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-33472045

RESUMO

Asthenoteratozoospermia characterized by multiple morphological abnormalities of the flagella (MMAF) has been identified as a sub-type of male infertility. Recent progress has identified several MMAF-associated genes with an autosomal recessive inheritance in human affected individuals, but the etiology in approximately 40% of affected individuals remains unknown. Here, we conducted whole-exome sequencing (WES) and identified hemizygous missense variants in the X-linked CFAP47 in three unrelated Chinese individuals with MMAF. These three CFAP47 variants were absent in human control population genome databases and were predicted to be deleterious by multiple bioinformatic tools. CFAP47 encodes a cilia- and flagella-associated protein that is highly expressed in testis. Immunoblotting and immunofluorescence assays revealed obviously reduced levels of CFAP47 in spermatozoa from all three men harboring deleterious missense variants of CFAP47. Furthermore, WES data from an additional cohort of severe asthenoteratozoospermic men originating from Australia permitted the identification of a hemizygous Xp21.1 deletion removing the entire CFAP47 gene. All men harboring hemizygous CFAP47 variants displayed typical MMAF phenotypes. We also generated a Cfap47-mutated mouse model, the adult males of which were sterile and presented with reduced sperm motility and abnormal flagellar morphology and movement. However, fertility could be rescued by the use of intra-cytoplasmic sperm injections (ICSIs). Altogether, our experimental observations in humans and mice demonstrate that hemizygous mutations in CFAP47 can induce X-linked MMAF and asthenoteratozoospermia, for which good ICSI prognosis is suggested. These findings will provide important guidance for genetic counseling and assisted reproduction treatments.


Assuntos
Astenozoospermia/genética , Infertilidade Masculina/genética , Animais , Astenozoospermia/patologia , Astenozoospermia/fisiopatologia , Estudos de Coortes , Feminino , Deleção de Genes , Genes Ligados ao Cromossomo X , Hemizigoto , Humanos , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Infertilidade Masculina/fisiopatologia , Masculino , Camundongos Endogâmicos C57BL , Mutação , Mutação de Sentido Incorreto , Linhagem , Fenótipo , Injeções de Esperma Intracitoplásmicas , Motilidade Espermática , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/patologia , Espermatozoides/fisiologia , Espermatozoides/ultraestrutura , Sequenciamento Completo do Exoma
2.
Am J Hum Genet ; 107(3): 514-526, 2020 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-32791035

RESUMO

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenoteratozoospermia. Although recent studies have revealed several MMAF-associated genes and demonstrated MMAF to be a genetically heterogeneous disease, at least one-third of the cases are still not well understood for their etiology. Here, we identified bi-allelic loss-of-function variants in CFAP58 by using whole-exome sequencing in five (5.6%) unrelated individuals from a cohort of 90 MMAF-affected Chinese men. Each of the men harboring bi-allelic CFAP58 variants presented typical MMAF phenotypes. Transmission electron microscopy demonstrated striking flagellar defects with axonemal and mitochondrial sheath malformations. CFAP58 is predominantly expressed in the testis and encodes a cilia- and flagella-associated protein. Immunofluorescence assays showed that CFAP58 localized at the entire flagella of control sperm and predominantly concentrated in the mid-piece. Immunoblotting and immunofluorescence assays showed that the abundances of axoneme ultrastructure markers SPAG6 and SPEF2 and a mitochondrial sheath protein, HSP60, were significantly reduced in the spermatozoa from men harboring bi-allelic CFAP58 variants. We generated Cfap58-knockout mice via CRISPR/Cas9 technology. The male mice were infertile and presented with severe flagellar defects, consistent with the sperm phenotypes in MMAF-affected men. Overall, our findings in humans and mice strongly suggest that CFAP58 plays a vital role in sperm flagellogenesis and demonstrate that bi-allelic loss-of-function variants in CFAP58 can cause axoneme and peri-axoneme malformations leading to male infertility. This study provides crucial insights for understanding and counseling of MMAF-associated asthenoteratozoospermia.


Assuntos
Anormalidades Múltiplas/genética , Astenozoospermia/genética , Axonema/genética , Infertilidade Masculina/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Anormalidades Múltiplas/patologia , Alelos , Animais , Astenozoospermia/fisiopatologia , Axonema/patologia , Sistemas CRISPR-Cas/genética , Proteínas de Ciclo Celular/genética , Homozigoto , Humanos , Infertilidade Masculina/patologia , Mutação com Perda de Função/genética , Perda de Heterozigosidade/genética , Masculino , Camundongos , Camundongos Knockout , Proteínas dos Microtúbulos/genética , Mitocôndrias/genética , Cauda do Espermatozoide/metabolismo , Cauda do Espermatozoide/patologia , Testículo/metabolismo , Testículo/patologia , Sequenciamento Completo do Exoma
3.
Acupunct Med ; 37(1): 25-32, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30942613

RESUMO

OBJECTIVE: To evaluate the effect of transcutaneous electrical acupuncture point stimulation (TEAS) on sperm parameters and the underlying molecular mechanisms. METHODS: A total of 121 patients diagnosed with oligozoospermia, asthenozoospermia or oligoasthenozoospermia were randomised into four groups (three treatment groups, one control): the TEAS groups were treated with 2 Hz (n=31), 100 Hz (n=31), or mock stimulation (n=29) at acupuncture points BL23, ST36, CV1 and CV4 for 2 months. The control group (n=30) was provided with lifestyle advice only. RESULTS: The changes in total sperm count and motility in the 2 Hz TEAS group were significantly greater than those in the mock group and the control group. The change in neutral α-glucosidase (NAG) and zinc levels in the 2 Hz group were significantly greater than those in the mock group and control group, and the changes in fructose levels of the 2 Hz group were significantly greater than those in the control group. Significant increases in calcium and integrin-binding protein 1 (CIB1) and reduction of cyclin-dependent kinase 1 b (CDK1) were also found after 2 Hz TEAS treatment. CONCLUSIONS: The present findings suggest that 2 Hz TEAS can improve sperm count and motility in patients with abnormal semen parameters, and is associated with increases in seminal plasma zinc, NAG and fructose. The upregulation of CIB1 and downregulation of CDK1 by TEAS may be associated with its positive effects on sperm motility and count. TRIAL REGISTRATION: http://www.chictr.org ; registration no. ChiCTR-TRC-11001775.


Assuntos
Astenozoospermia/terapia , Eletroacupuntura , Oligospermia/terapia , Sêmen/metabolismo , Pontos de Acupuntura , Adulto , Astenozoospermia/metabolismo , Astenozoospermia/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Oligospermia/metabolismo , Oligospermia/fisiopatologia , Sêmen/citologia , Contagem de Espermatozoides , Motilidade Espermática , Espermatozoides/citologia , Resultado do Tratamento
4.
Fertil Steril ; 111(5): 897-908.e2, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31029245

RESUMO

OBJECTIVE: To characterize in depth and investigate the role of exosomes present in seminal plasma in affecting parameters underlying sperm activity. DESIGN: In vitro experimental study. SETTING: Research hospital. PATIENT(S): Normozoospermic, severe asthenozoospermic, and post-vasectomy azoospermic men 18-55 years of age were considered for the study. Seminal plasma was collected and processed to separate spermatozoa and exosomes. INTERVENTION(S): None. MAIN OUTCOMES MEASURE(S): Exosomes from seminal plasma were isolated and characterized by means of nanoparticle tracking analysis, transmission electron microscopy and Western blot. Exosome uptake by spermatozoa was monitored by means of immunofluorescence and flow cytometry. The effect of exosomes on spermatozoa was determined by evaluating progressive motility and capacitation, the latter assessed by means of tyrosine phosphorylation and acrosome reaction. RESULT(S): We isolated and characterized exosomes from seminal plasma of normo-, astheno-, and azoospermic patients. They display similar features in terms of shape, size, expression of canonic exosome markers and proteins involved in spermatozoa maturation, and fertilization capacity. After ejaculation, sperm cells are still receptive and are able to take up exosomes in a time- and pH-dependent manner. Exosomes derived from normozoospermic but not from asthenozoospermic individuals improve spermatozoa motility and trigger capacitation. Transfer of cysteine-rich secretory protein 1 from exosomes to spermatozoa may have a role in these phenomena. CONCLUSION(S): These findings provide evidence that: 1) sperm can still receive vesicle-derived cargo after ejaculation; 2) sperm motility and ability to undergo capacitation can benefit from exosomal transfer; and 3) semen quality is affected by male tract exosomes.


Assuntos
Astenozoospermia/diagnóstico , Exossomos/fisiologia , Sêmen/fisiologia , Capacitação Espermática/fisiologia , Motilidade Espermática/fisiologia , Espermatozoides/fisiologia , Adolescente , Adulto , Astenozoospermia/genética , Astenozoospermia/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Adulto Jovem
5.
Open Biol ; 9(4): 180091, 2019 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-31014201

RESUMO

Asthenozoospermia is a common cause of male infertility, the aetiology of which remains unclear in 50-60% of cases. The current study aimed to characterize metabolic alterations in asthenozoospermic seminal plasma and to explore the signalling pathways involved in sperm motility regulation. At first, high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry was used to detect the targeted metabolic network of arachidonic acid (AA). Metabolomic multivariate data analysis showed significant distinction of AA metabolites between asthenozoospermic and healthy seminal plasma. AA as well as its lipoxygenase (LOX) and cytochrome P450 metabolites were found to be abnormally increased, while cyclooxygenase (COX) metabolites were complicatedly disturbed in asthenozoospermic volunteers compared with those in healthy ones. In vitro experiments and western blot analysis of sperm cells revealed a decrease in sperm motility and upregulation of sperm phosphor-P38 induced by AA. P38 inhibitor could increase AA-reduced sperm motility. Also, all the inhibitors of the three metabolic pathways of AA could block AA-induced P38 mitogen-activated protein kinase (MAPK) activation and further improve sperm motility. We report here for the first time that an abnormal AA metabolic network could reduce sperm motility via P38 MAPK activation through the LOX, cytochrome P450 and COX metabolic pathways, which might be an underlying pathomechanism of asthenozoospermia.


Assuntos
Ácido Araquidônico/metabolismo , Redes e Vias Metabólicas/fisiologia , Motilidade Espermática/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Adulto , Astenozoospermia/metabolismo , Astenozoospermia/fisiopatologia , Cromatografia Líquida de Alta Pressão/métodos , Sistema Enzimático do Citocromo P-450/metabolismo , Humanos , Lipoxigenase/metabolismo , Masculino , Metabolômica/métodos , Pessoa de Meia-Idade , Prostaglandina-Endoperóxido Sintases/metabolismo , Transdução de Sinais , Espermatozoides/metabolismo , Espermatozoides/fisiologia , Espectrometria de Massas em Tandem/métodos , Adulto Jovem
6.
Mol Hum Reprod ; 25(4): 171-183, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30824926

RESUMO

In mouse and bovine sperm, GSK3 activity is inversely proportional to motility. Targeted disruption of the GSK3A gene in testis results in normal spermatogenesis, but mature sperm present a reduced motility, rendering male mice infertile. On the other hand, GSK3B testis-specific KO is fertile. Yet in human sperm, an isoform-specific correlation between GSK3A and sperm motility was never established. In order to analyze GSK3 function in human sperm motility, normospermic and asthenozoospermic samples from adult males were used to correlate GSK3 expression and activity levels with human sperm motility profiles. Moreover, testicular and sperm GSK3 interactomes were identified using a yeast two-hybrid screen and coimmunoprecipitation, respectively. An extensive in-silico analysis of the GSK3 interactome was performed. The results proved that inhibited GSK3A (serine phosphorylated) presents a significant strong positive correlation (r = 0.822, P = 0.023) with the percentage of progressive human sperm, whereas inhibited GSK3B is not significantly correlated with sperm motility (r = 0.577, P = 0.175). The importance of GSK3 in human sperm motility was further reinforced by in-silico analysis of the GSK3 interactome, which revealed a high level of involvement of GSK3 interactors in sperm motility-related functions. The limitation of techniques used for GSK3 interactome identification can be a drawback, since none completely mimics the physiological environment. Our findings prove that human sperm motility relies on isoform-specific functions of GSK3A within this cell. Given the reported relevance of GSK3 protein-protein interactions in sperm motility, we hypothesized that they stand as potential targets for male contraceptive strategies based on sperm motility modulation.


Assuntos
Astenozoospermia/genética , Fertilidade/genética , Quinase 3 da Glicogênio Sintase/genética , Processamento de Proteína Pós-Traducional , Motilidade Espermática/genética , Espermatogênese/genética , Espermatozoides/enzimologia , Adulto , Animais , Astenozoospermia/enzimologia , Astenozoospermia/fisiopatologia , Bovinos , Expressão Gênica , Perfilação da Expressão Gênica , Quinase 3 da Glicogênio Sintase/metabolismo , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Camundongos , Fosforilação , Ligação Proteica , Mapeamento de Interação de Proteínas , Espermatozoides/patologia , Testículo/enzimologia , Testículo/patologia
7.
Life Sci ; 218: 81-88, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30550884

RESUMO

AIMS: Idiopathic nature of male infertility disorder needs to be investigated by different horizons of molecular biology for its treatment and to device male contraceptive. Further, it can also aid in advancement of assisted reproductive technology (ART), as nowadays the failure and disquiets of ART are consistent. Herein, we have attempted to find out proteins responsible for male infertility by comparing proteome profile of sperms collected from normal control and asthenozoospermic (AS) males. MAIN METHODS: Differential proteome profiles were studied by 2-dimensional differential gel electrophoresis (2D-DIGE) and mass spectrometry. The confirmation of proteome profiling results was done by western blotting and ELISA. Quantitative reverse-transcription-PCR was also performed in an independent cohort of AS and normal individuals to investigate the transcriptional regulation of proteins. KEY FINDINGS: Although seven differentially regulated proteins were identified, highpoints of the study were two proteins, TEX40 and ATP6V0A2. Lower expression of a crucial sperm motility related protein, TEX40 is reported for the first time in clinically diagnosed AS males in the present investigation. Most likely with reference to previous findings the down regulation of TEX40 leads to fewer entries of calcium ions in the sperm and lower expression of ATP6V0A2 is responsible for acrosomal de-acidification. SIGNIFICANCE: Conclusively, the down regulation of these two proteins in AS males might result in diminished sperm motility. The findings can be worthwhile for male contraception and ART management besides their use for male infertility therapy.


Assuntos
Acrossomo/metabolismo , Astenozoospermia/fisiopatologia , Canais de Cálcio/metabolismo , Cálcio/metabolismo , Proteômica/métodos , ATPases Translocadoras de Prótons/metabolismo , Espermatozoides/metabolismo , Adulto , Astenozoospermia/metabolismo , Biomarcadores/metabolismo , Estudos de Casos e Controles , Humanos , Masculino , Prognóstico
8.
Reprod Sci ; 26(7): 997-1004, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30270743

RESUMO

Given the higher risk of developing imprinting disorders in assisted reproductive technology (ART)-conceived children, we hypothesized that ART may affect DNA methylation of the insulin-like growth factor 2 (IGF2), H19, small nuclear ribonucleoprotein polypeptide N (SNRPN) differentially methylated regions (DMRs) at the fetal stage, which in turn may be associated with sperm abnormalities. A total of 4 patient groups were recruited, namely, multifetal reduction following in vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI; n = 56), multifetal reduction following controlled ovarian hyperstimulation (COH; n = 42), male patients with normal semen parameters denoted as normozoospermia group (NZ) for IVF (n = 36), and male patients presenting with asthenozoospermia (OAZ) for ICSI (n = 38). The expression levels and the DNA methylation status of IGF2-H19 and SNRPN DMRs in the fetuses and the semen samples were evaluated by real-time quantitative polymerase chain reaction and pyrosequencing. In our results, the expression levels of H19 were significantly higher, whereas the methylation rates were lower in IVF-conceived fetuses compared to the control group (P < .05). Furthermore, higher methylation rates of IGF2 DMR2 and SNRPN DMR were detected both in IVF- and ICSI-conceived fetuses (P < .05). The data further indicated that the patients who presented with the majority of the CpG sites in the H19 DMR region that were lower methylated were those in the OAZ group. The results demonstrated that the epigenetic dysregulations of IGF2-H19 and SNRPN DMRs that were caused by ART were noted in the fetuses. Moreover, the present study suggested that epigenetic perturbations of the H19 DMR might be a key biomarker for spermatogenesis defects in humans.


Assuntos
Astenozoospermia/genética , Metilação de DNA , Fertilização In Vitro/efeitos adversos , Feto/metabolismo , Loci Gênicos , Impressão Genômica , Fator de Crescimento Insulin-Like II/genética , RNA Longo não Codificante/genética , Espermatozoides/metabolismo , Adulto , Astenozoospermia/metabolismo , Astenozoospermia/patologia , Astenozoospermia/fisiopatologia , Estudos de Casos e Controles , Ilhas de CpG , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Fator de Crescimento Insulin-Like II/metabolismo , Masculino , Gravidez , RNA Longo não Codificante/metabolismo , Injeções de Esperma Intracitoplásmicas/efeitos adversos , Espermatogênese/genética , Espermatozoides/patologia , Proteínas Centrais de snRNP/genética , Proteínas Centrais de snRNP/metabolismo
9.
Andrologia ; 51(1): e13146, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30255511

RESUMO

Maintaining sperm motility after ejaculation is important for fertilisation. Apoptosis may play an important role to reduce sperm motility after ejaculation. The aim of this study was to perceive whether or not an increase in apoptosis reduces sperm motility in a higher degree after ejaculation and whether it can be predicted by laboratory tests, such as sperm chromatin structure assay (SCSA). Fifty-one Asthenozoospermia and 20 fertile subjects participated in this study. SCSA was applied using flow cytometry. Fluorescein-labelled inhibitors of Caspases (FLICA) method was used for assessment of active Caspase-3. Motility was assessed every 2 hr after ejaculation for 12 hr. Both SCSA and spermatozoa with active Caspase-3 were significantly correlated with the rate of motility reduction after ejaculation. In the subgroups who had SCSA <27% and active Caspase-3 <40%, the sperm motility reduction significantly occurred 6-8 hr after ejaculation compared to the fresh sample. In the cases of SCSA ≥27% and active Caspase-3 ≥ 40%, a significant decrease in motility was observed between 2 and 4 hr after ejaculation. The result demonstrated a significant trend in the rate of sperm motility reduction with SCSA increase, which suggests SCSA may indirectly show a good scheme of apoptosis status and may forecast the rate of motility reduction after ejaculation in Asthenozoospermia.


Assuntos
Astenozoospermia/fisiopatologia , Cromatina/metabolismo , Motilidade Espermática/fisiologia , Espermatozoides/fisiologia , Adulto , Apoptose/fisiologia , Astenozoospermia/metabolismo , Caspase 3/metabolismo , Dano ao DNA/fisiologia , Fragmentação do DNA , Ejaculação/fisiologia , Humanos , Masculino , Espermatozoides/metabolismo , Adulto Jovem
10.
Isr Med Assoc J ; 20(4): 250-253, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29629734

RESUMO

BACKGROUND: Male infertility is solely responsible for approximately 20% of all infertility in couples. Various factors have been proposed as having a negative effect on sperm quality; however, the reasons for the global decline in sperm parameters during the last few decades are still controversial. OBJECTIVES: To investigate the fluctuations of semen parameters (sperm concentration, motility, and morphology) in three sperm quality groups and to examine the trends of those parameters in the same men over time. RESULTS: Our data showed deterioration in all semen parameters assessed in the group of men originally considered as having normal semen values according to the 2010 criteria of the World Health Organization. In contrast, we found significant improvement over time in all semen parameters in the group of men with severe oligo-terato-asthenozoospermia. CONCLUSIONS: Our results suggest that, although there were changes in sperm quality over time in the groups assessed, the clinical significance is negligible and does not necessarily justify a change in the therapeutic approach to infertility or sperm cryopreservation.


Assuntos
Infertilidade Masculina/fisiopatologia , Sêmen/fisiologia , Contagem de Espermatozoides/métodos , Motilidade Espermática/fisiologia , Espermatozoides/fisiologia , Astenozoospermia/fisiopatologia , Estudos de Coortes , Seguimentos , Humanos , Masculino , Oligospermia/fisiopatologia , Estudos Retrospectivos , Índice de Gravidade de Doença , Teratozoospermia/fisiopatologia , Fatores de Tempo
11.
J Cell Physiol ; 233(1): 11-22, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28247940

RESUMO

Reduced sperm motility (asthenospermia) and resulting infertility arise from deletion of the Plasma Membrane Ca2+ -ATPase 4 (Pmca4) gene which encodes the highly conserved Ca2+ efflux pump, PMCA4. This is the major Ca2+ clearance protein in murine sperm. Since the mechanism underlying asthenospermia in PMCA4's absence or reduced activity is unknown, we investigated if sperm PMCA4 negatively regulates nitric oxide synthases (NOSs) and when absent NO, peroxynitrite, and oxidative stress levels are increased. Using co-immunoprecipitation (Co-IP) and Fluorescence Resonance Energy Transfer (FRET), we show an association of PMCA4 with the NOSs in elevated cytosolic [Ca2+ ] in capacitated and Ca2+ ionophore-treated sperm and with neuronal (nNOS) at basal [Ca2+ ] (ucapacitated sperm). FRET efficiencies for PMCA4-eNOS were 35% and 23% in capacitated and uncapacitated sperm, significantly (p < 0.01) different, with the molecules being <10 nm apart. For PMCA4-nNOS, this interaction was seen only for capacitated sperm where FRET efficiency was 24%, significantly (p < 0.05) higher than in uncapacitated sperm (6%). PMCA4 and the NOSs were identified as interacting partners in a quaternary complex that includes Caveolin1, which co-immunoprecipitated with eNOS in a Ca2+ -dependent manner. In Pmca4-/- sperm NOS activity was elevated twofold in capacitated/uncapacitated sperm (vs. wild-type), accompanied by a twofold increase in peroxynitrite levels and significantly (p < 0.001) increased numbers of apoptotic germ cells. The data support a quaternary complex model in which PMCA4 co-ordinates Ca2+ and NO signaling to maintain motility, with increased NO levels resulting in asthenospermia in Pmca4-/- males. They suggest the involvement of PMCA4 mutations in human asthenospermia, with diagnostic relevance.


Assuntos
Astenozoospermia/enzimologia , Sinalização do Cálcio , ATPases Transportadoras de Cálcio/metabolismo , Membrana Celular/enzimologia , Óxido Nítrico/metabolismo , Motilidade Espermática , Espermatozoides/enzimologia , Animais , Apoptose , Astenozoospermia/genética , Astenozoospermia/patologia , Astenozoospermia/fisiopatologia , ATPases Transportadoras de Cálcio/deficiência , ATPases Transportadoras de Cálcio/genética , Caveolina 1/metabolismo , Fertilidade , Transferência Ressonante de Energia de Fluorescência , Predisposição Genética para Doença , Masculino , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Camundongos Knockout , Complexos Multienzimáticos , Óxido Nítrico Sintase Tipo I/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Estresse Oxidativo , Ácido Peroxinitroso/metabolismo , Fenótipo , Espermatozoides/patologia
12.
Rom J Intern Med ; 56(1): 15-20, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-28865234

RESUMO

INTRODUCTION: Human seminal plasma contains a variety of macro and trace elements including magnesium (Mg), copper (Cu), zinc (Zn), and iron (Fe) that have essential roles in normal functioning of semen and its quality. The imbalance of these elements has been reported in several pathologic and male infertility disorders. Therefore, this study aimed to determine the levels of these elements in seminal plasma samples, their relationships with each other and their impact on sperm motility. METHODS: Overall, 192 males (96 normospermic and 96 asthenospermic males) were enrolled in the study. Semen samples were collected by masturbation and computer-assisted/aided semen analysis of sperm motility was performed. The samples were centrifuged and seminal levels of Mg, Cu, Zn and Fe were measured using atomic absorption spectroscopy. RESULTS: The levels of Zn did not differ between the two groups, while the levels of Mg, Cu, and Fe were significantly higher in normospermic males. Fe showed a positive correlation with Mg and Cu in asthenospermic group. However, a negative relationship was found between Mg and Fe levels and between Mg and sperm concentration in the normospermic group. Fe levels were higher in the normospermic group compared to the asthenospermic group. Nevertheless, increased Fe levels caused a decrease in most of sperm motility fractions. CONCLUSION: Elements play major roles in male fertility and directly affect sperm quality. According to the results of this study, the levels of Zn do not affect the sperm quality and motility, while Fe, Cu and Mg are decreased in males with sperm motility problems. Nevertheless, Fe levels can adversely affect sperm motility in normospermic men.


Assuntos
Astenozoospermia/fisiopatologia , Sêmen/metabolismo , Motilidade Espermática/fisiologia , Oligoelementos/metabolismo , Astenozoospermia/metabolismo , Cobre/análise , Cobre/metabolismo , Humanos , Ferro/análise , Ferro/metabolismo , Magnésio/análise , Magnésio/metabolismo , Masculino , Sêmen/química , Oligoelementos/análise , Zinco/análise , Zinco/metabolismo
13.
Andrologia ; 50(2)2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28762516

RESUMO

To study the outcome of FISH sperm examination in cases with sperm pathology and outline the potential correlation with certain chromosomal defects. A retrospective study of prospectively collected data was performed in IAKENTRO, Infertility Treatment Center. Rates of abnormal FISH semen examination were compared between male infertility patients and fertile controls. Detection of abnormal FISH semen examination as well as each chromosomal abnormality detected was correlated with each sperm deficiency (asthenozoospermia, oligozoospermia and teratozoospermia) in a univariate regression model. There were 72 male partners included, of which 52 male infertility patients and 20 controls. The rate of abnormal sperm FISH examination was significantly higher in patients' group (55.8% vs. 15.0% for controls, p = .002). Asthenozoospermia, oligozoospermia and teratozoospermia were significantly correlated with detection of abnormal FISH examination (p = .004, p = .01 and p < .001 respectively). Teratospermia was significantly correlated with increased aneuploidy rate for chromosome 17 (p = .005), chromosome X (p = .05) and Y (p = .03). FISH examination reveals pathology in a significant proportion of patients with sperm defects and should be recommended to achieve early detection of chromosomal defects that may postpone favourable reproductive outcome.


Assuntos
Astenozoospermia/fisiopatologia , Oligospermia/fisiopatologia , Análise do Sêmen/métodos , Espermatozoides/patologia , Teratozoospermia/fisiopatologia , Adulto , Aneuploidia , Astenozoospermia/genética , Estudos de Casos e Controles , Cromossomos Humanos Par 17/genética , Cromossomos Humanos X/genética , Cromossomos Humanos Y/genética , Humanos , Hibridização in Situ Fluorescente , Masculino , Oligospermia/genética , Estudos Prospectivos , Estudos Retrospectivos , Teratozoospermia/genética
14.
Andrologia ; 50(2)2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28782302

RESUMO

Sperm DNA damage, excessive oxidative stress and decrease in motility may lead to low fertilisation or poor assisted reproductive techniques outcomes in asthenoteratozoospermic men. Selenium was considered as essential element for male reproductive functions. Selenium has important role in enzymatic process for elimination of excessive reactive oxygen species and helps to maintain membrane integrity. The aim of this study was to determine the effect of selenium supplementation on sperm quality, DNA fragmentation, mitochondrial membrane potential and membrane lipid peroxidation during sperm sampling in vitro at different times. In this experimental study, semen samples were collected from 50 asthenoteratozoospermic men. Samples were divided into two groups as control group and test group (incubated with 2 µg/ml selenium at 37°C for 2, 4 and 6 hr). Motility and viability were assessed based on WHO 2010 criteria. Mitochondrial membrane potential, sperm DNA fragmentation and malondialdehyde levels were evaluated in each group. Results revealed that motility, viability and mitochondrial membrane potential were significantly higher in the test group (p < .05). Also malondialdehyde levels were significantly lower in the test group (p < .03). DNA fragmentation significantly decreased in the test group after 6 hr of incubation (p < .02). In conclusion, in vitro selenium supplementation may protect spermatozoa from maltreatment effect of reactive oxygen species (ROS) during sperm sampling via keeping enzymatic and antioxidant process in optimum condition.


Assuntos
Antioxidantes/farmacologia , Astenozoospermia/terapia , Técnicas de Reprodução Assistida , Selênio/farmacologia , Espermatozoides/efeitos dos fármacos , Adulto , Astenozoospermia/fisiopatologia , Fragmentação do DNA/efeitos dos fármacos , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Análise do Sêmen , Motilidade Espermática/efeitos dos fármacos , Recuperação Espermática/efeitos adversos , Espermatozoides/fisiologia , Adulto Jovem
15.
Biochim Biophys Acta Gen Subj ; 1862(3): 660-668, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29247744

RESUMO

BACKGROUND: We previously cloned the Ssp411 gene. We found that the Ssp411 protein is predominantly expressed in elongated spermatids in the rat testis in a stage-dependent manner. Although our findings strongly suggested that Ssp411 might play an important role in mammalian spermatogenesis, this hypothesis has not been studied. METHODS: We first used real-time PCR, Western blotting and immunohistochemistry to confirm that the expression pattern of Ssp411 in several murine tissues is similar to its expression pattern in corresponding rat tissues. To better understand the roles of Ssp411 in male reproduction in vivo, we identified and characterized an Ssp411 expression-disrupted murine strain (Ssp411PB/PB) that was generated by piggyBac (PB) transposon insertion. We studied Ssp411-interacting proteins using proteome microarray, co-IP and GST pull-down assay. RESULTS: Both Ssp411 mRNA and protein were detected exclusively in spermatids after step 9 during spermiogenesis in testis. Phenotypic analysis suggested that only Ssp411PB/PB males are sterile. These males have smaller testes, reduced sperm counts, decreased sperm motility and deformed spermatozoa. Microscopy analysis indicated that the manchette, a structurally reshaped sperm head, is aberrant in Ssp411PB/PB spermatids. The results of proteome microarray analysis and GST pull-down assays suggested that Ssp411 participates the ubiquitin-proteasome system by interacting with PSMC3. This has been reported to be manchette-associated and important for the head shaping of spermatids. CONCLUSIONS: Our study suggested that Ssp411 is required for spermiogenesis. It seems to play a role in sperm head shaping. The lack of Ssp411 causes sperm deformation and results in male infertility. GENERAL SIGNIFICANCE: Ssp411PB/PB mouse strain is an animal model of idiopathic oligoasthenoteratozoospermia (iOAT), and the gene may represent a therapeutic target for iOAT patients.


Assuntos
Astenozoospermia/genética , Cabeça do Espermatozoide/ultraestrutura , Espermatogênese/fisiologia , Teratozoospermia/genética , Animais , Astenozoospermia/fisiopatologia , Elementos de DNA Transponíveis , Modelos Animais de Doenças , Feminino , Humanos , Masculino , Camundongos , Camundongos Mutantes , Microtúbulos/patologia , Mutagênese Insercional , Especificidade de Órgãos , Complexo de Endopeptidases do Proteassoma/metabolismo , Mapeamento de Interação de Proteínas , Proteoma , RNA Mensageiro/biossíntese , Motilidade Espermática , Espermátides/metabolismo , Espermatogênese/genética , Teratozoospermia/fisiopatologia , Testículo/metabolismo , Ubiquitina/metabolismo
16.
Syst Biol Reprod Med ; 64(2): 112-121, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29083935

RESUMO

Androgen mediating signaling is implicated in regulating the expression of reproductive related genes. Any deviation in the gene expression might be the ignitable precursor for genomic instability that inflames the genomic rearrangements and a leading cause of cancer. The main goal of this study was to determine the relationships between trinucleotide repeats of androgen receptor (AR), sex steroids, and sexual function in men presenting with reduced sperm motility. We investigated the singleton or combinatorial effects of the length of trinucleotide repeats of AR on reproductive hormones, clinical parameters, semen analyses, as well as sexual assessment function of 210 asthenospermic outpatients along with 125 normal subjects. Sexual assessment was executed using the International Index of Erectile Function (IIEF-15 score) which measures erectile function (EF), orgasmic function (OR), sexual desire (SD), intercourse satisfaction (IS), and overall satisfaction. Our findings suggest that long (>26 CAG)n repeats have an inverse correlation with circulatory FSH and T, whereas long (>25 GGC)n repeats have moderated affiliation with reduced sperm concentration. The study revealed a novel finding by exploring the negative correlation between elongated (CAG)n repeats and the cumulative IIEF-15 score, orgasm function (OR), and erectile function (EF) in asthenospermic men. This study examines the tri-nucleotide correlation with sexual function in Punjabi men enhancing our understanding of the regulatory mechanisms of sexual performance. ABBREVIATIONS: AR: androgen receptor; IIEF-15 score: International Index of Erectile Function; EF: erectile function; OR: orgasmic function; SD: sexual desire; IS: intercourse satisfaction; FSH: follicular stimulating hormone; T: testosterone; NTD: N-terminal transactivation domain; DBD: DNA-binding domain; LBD: ligand binding domain; TNR: tri-nucleotide repeat.


Assuntos
Astenozoospermia/sangue , Astenozoospermia/genética , Hormônio Foliculoestimulante Humano/sangue , Receptores Androgênicos/genética , Testosterona/sangue , Repetições de Trinucleotídeos , Adulto , Astenozoospermia/fisiopatologia , Biomarcadores/sangue , Estudos de Casos e Controles , Fertilidade , Humanos , Masculino , Pessoa de Meia-Idade , Paquistão , Reprodução , Comportamento Sexual , Motilidade Espermática
17.
J Huazhong Univ Sci Technolog Med Sci ; 37(6): 915-921, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29270753

RESUMO

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations (group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control (group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate (CR), tail DNA percentage (TD), tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.


Assuntos
Acetilcarnitina/farmacologia , Acrossomo/efeitos dos fármacos , Antioxidantes/farmacologia , Astenozoospermia/fisiopatologia , Criopreservação/métodos , Crioprotetores/farmacologia , Acrossomo/metabolismo , Acrossomo/ultraestrutura , Astenozoospermia/metabolismo , Estudos de Casos e Controles , Sobrevivência Celular/efeitos dos fármacos , Humanos , Infertilidade Masculina/metabolismo , Infertilidade Masculina/fisiopatologia , Masculino , Estresse Oxidativo , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Motilidade Espermática/efeitos dos fármacos
18.
Zhen Ci Yan Jiu ; 42(2): 114-8, 2017 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-29071957

RESUMO

OBJECTIVE: To investigate the effects of different electroacupuncture (EA) parameters for the treatment of asthenozoospermia in rats. METHODS: One hundred and five male Sprague-Dawley rats were randomly divided into 2 Hz-EA treatment daily in 3 d group (n=9), sham-EA group (n=10), model group (n=10); 2 Hz-EA treatment every other day in 5 d group, sham-EA group, model group (8 rats in each group); 2 Hz-EA treatment every other day in 9 d group, sham-EA group, model group (10 rats in each group); 100 Hz-EA treatment every other day in 9 d group (n=7), sham-EA group (n=8), model group (n=7). Asthenozoospermia model was established by intragastric administration of ornidazole (ORN,400 mg·kg-1·d-1) once daily till the end of treatment. EA treatments (2 Hz or 100 Hz) were applied to "Shenshu" (BL 23,bilateral), "Zusanli" (ST 36, bilateral) for 30 min, intensity of 1-2-3 mA (increasing 1 mA per 10 min), once a day or once every other day for 3 times or 5 times. Sham-EA groups were treated with similar procedure except that the output leads of the stimulator were disconnected. The sperm density, viability, motility, the number of grade A sperm, and grade A+B sperm were examined by computer-assisted sperm analysis. RESULTS: (1) 2 Hz-EA treatment daily in 3 d:compared with the model group and the sham-EA group, 2 Hz-EA treatment once daily had no significant effect on all of the sperm motility indexes in the asthenozoospermic rats (P>0.05). (2) 2 Hz-EA treatment every other day in 5 d:compared with the model group, EA treatment could increase the sperm motility (P<0.05), the number of grade A sperm (P<0.05), and the number of grade A+B sperm (P<0.05) in the asthenozoospermic rats. However, compared with the sham-EA group, EA treatment could only improve the number of grade A+B sperm (P<0.05). (3) 2 Hz-EA treatment every other day in 9 d:compared with both the model group and the sham-EA group, EA treatment could markedly improve the sperm viability (P<0.001), the sperm motility (P<0.001), the number of grade A sperm (P<0.001), and the number of grade A+B sperm (P<0.001) in the asthenozoospermic rats. (4) 100 Hz-EA treatment every other day in 9 days:compared with both the model group and the sham-EA group, all of the sperm indexes in the asthenozoospermic rats including the sperm viability (P<0.001 vs. the model group, P<0.05 vs. the sham-EA group), the sperm motility (P<0.001 vs. the model group, P<0.01 vs. the sham-EA group), the number of grade A sperm (P<0.01) and the number of grade A+B sperm (P<0.01) also could be improved after EA treatment. Unexpectedly,none of the EA treatment had significant influence on the sperm density in the asthenozoospermic rats. CONCLUSIONS: Both 2 Hz-EA and 100 Hz-EA treatment once every other day for 5 times in 9 d had a therapeutic effect on asthenozoospermia by improving the sperm viability and the sperm motility in the rats.


Assuntos
Pontos de Acupuntura , Astenozoospermia/terapia , Animais , Astenozoospermia/fisiopatologia , Eletroacupuntura , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Contagem de Espermatozoides , Espermatozoides/citologia
19.
Arch Gynecol Obstet ; 296(3): 589-596, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28707057

RESUMO

PURPOSE: To evaluate the effect of different concentrations and durations of seminal testosterone supplementation upon the motility of sperm from asthenozoospermic males. METHODS: Semen was collected from 41 infertile men with asthenozoospermia. After liquefaction, 200 µL was extracted from each semen sample and divided equally into five groups for a negative control, a vehicle control, and three experimental portions mixed with 4.75, 7.75, and 17.75 nmol/L of testosterone, respectively. The sperm motility was evaluated at 5, 15, 30, and 45 min following the addition of testosterone. The supernatant from remaining samples were sent for testosterone assay. Sperm viability was also evaluated after 45 min. RESULTS: There was no difference in the number of samples in each group which showed a 20% improvement in sperm motility. Group 3 showed a significant retardation in the reduction of motility compared with Group 5 (P < 0.05). Semen samples with a final testosterone concentration of 4.51-10 nmol/L showed a significant improvement in sperm motility when measured 5 min after addition. In contrast, samples showing a rise in testosterone level above 10 nmol/L were associated with a reduction in both sperm motility and viability. CONCLUSION: Despite sperm motility decreasing over time, supplementation of semen samples with 4.75 nmol/L of testosterone could delay such reduction. A final seminal testosterone concentration of 4.51-10 nmol/L appears to be optimal for the best sperm motility.


Assuntos
Astenozoospermia/fisiopatologia , Motilidade Espermática/efeitos dos fármacos , Testosterona/farmacologia , Humanos , Masculino , Análise do Sêmen
20.
Artigo em Inglês | MEDLINE | ID: mdl-28638871

RESUMO

BACKGROUND: Kidney tonifying - spleen strengthening method being one of the modalities for treatment of astheno-oligozoospermia is currently commonly used in the clinical setting. To investigate the mechanism of YiShenJianPi (YSJP) Recipe, used in Traditional Chinese Medicine to benefit "the kidney" and strengthen "the spleen". MATERIALS AND METHODS: Oligoasthenozoospermia, male BALB/c mice were randomly divided into normal control, disease model, positive control, low-dosage and high-dosage groups. Oligoasthenozoospermia was induced by tripterygium glucosides intragastric administration before treatment started. Through using computer-aided sperm analysis to test the changes in sperm quality, utilizing flow cytometry to test the percentage of sperm with normal mitochondrial transmembrane potential (JC-1 + %), utilizing X-ray microscopy to observe epididymal sperm ultra-microstructure placing special emphasis and photographing the differences in mitochondria of the flagellum region. RESULTS: Compared with DM, sperm quality of the treated mice was significantly better (P<0.05, respectively). Compared with PC, the LD group had significantly better quality sperms, while the parameters in the HD group were numerically better. Compared with NC, all other groups had significantly lower percentage of sperms with normal mitochondrial membrane potential. In PC, LD and HD groups, the percentage of sperms with normal mitochondrial membrane potential was significantly higher than that of D. The 9+9+2 mitochondrial sheath structure was complete in NC but damaged in DM. In the treatment groups, this structure was fairly clear. CONCLUSION: YSJP improved semen quality with oligoasthenozoospermia by improving sperm mitochondrial membrane potential and restoring sperm mitochondrial ultrastructure.


Assuntos
Astenozoospermia/tratamento farmacológico , Medicamentos de Ervas Chinesas/administração & dosagem , Tripterygium/toxicidade , Animais , Astenozoospermia/induzido quimicamente , Astenozoospermia/fisiopatologia , Glicosídeos/toxicidade , Humanos , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Mitocôndrias/efeitos dos fármacos , Motilidade Espermática , Espermatozoides/citologia , Espermatozoides/efeitos dos fármacos
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