RESUMO
To verify a possible synergistic effect of smoking and varicocele on the seminal plasma proteome and biological functions, a cross-sectional study was performed in 25 smokers and 24 nonsmokers. Samples were used for conventional semen analysis, functional analysis (DNA fragmentation, acrosome integrity and mitochondrial activity) and proteomics by a shotgun approach. Functional enrichment of biological pathways was performed in differentially expressed proteins. Smokers presented lower ejaculate volume (p = .027), percentage of progressively motile spermatozoa (p = .002), total sperm count (p = .039), morphology (p = .001) and higher percentage of immotile spermatozoa (p = .03), round cell (p = .045) and neutrophil count (p = .009). Smokers also presented lower mitochondrial activity and acrosome integrity and higher DNA fragmentation. We identified and quantified 421 proteins in seminal plasma, of which one was exclusive, 21 were overexpressed and 70 were underexpressed in the seminal plasma of smokers. The proteins neprilysin, beta-defensin 106A and histone H4A were capable of predicting the smoker group. Enriched functions were related to immune function and sperm machinery in testis/epididymis. Based on our findings, we can conclude that cigarette smoking leads to the establishment of inflammatory protein pathways in the testis/epididymis in the presence of varicocele that seems to act in synergy with the toxic components of the cigarette.
Assuntos
Fumar Cigarros/efeitos adversos , Infertilidade Masculina/imunologia , Sêmen/química , Proteínas de Plasma Seminal/análise , Varicocele/complicações , Acrossomo/efeitos dos fármacos , Acrossomo/imunologia , Acrossomo/patologia , Adulto , Brasil , Estudos Transversais , Fragmentação do DNA/efeitos dos fármacos , Epididimo/irrigação sanguínea , Epididimo/efeitos dos fármacos , Epididimo/imunologia , Humanos , Infertilidade Masculina/patologia , Masculino , Pessoa de Meia-Idade , não Fumantes/estatística & dados numéricos , Proteômica/estatística & dados numéricos , Sêmen/imunologia , Sêmen/metabolismo , Análise do Sêmen/estatística & dados numéricos , Proteínas de Plasma Seminal/metabolismo , Transdução de Sinais/imunologia , Fumantes/estatística & dados numéricos , Testículo/irrigação sanguínea , Testículo/efeitos dos fármacos , Testículo/imunologia , Nicotiana/toxicidade , Varicocele/imunologia , Adulto JovemRESUMO
High-fat diet is associated with hypercholesterolemia and seminal alterations in White New Zealand rabbits. We have previously reported disorders in the development of the manchette-acrosome complex during spermiogenesis and decreased testicular efficiency in hypercholesterolemic rabbits. On the other hand, olive oil incorporated into the diet improves cholesterolemia and semen parameters affected in hypercholesterolemic rabbits. In this paper, we report the recovery-with the addition of olive oil to diet-from the sub-cellular mechanisms involved in the shaping of the sperm cell and testicular efficiency altered in hypercholesterolemic rabbits. Using morphological (structural, ultra-structural and immuno-fluorescence techniques) and cell biology techniques, a reorganization of the manchette and related structures was observed when olive oil was added to the high-fat diet. Specifically, actin filaments, microtubules and lipid rafts-abnormally distributed in hypercholesterolemic rabbits-were recovered with dietary olive oil supplementation. The causes of the decline in sperm count were studied in the previous report and here in more detail. These were attributed to the decrease in the efficiency index and also to the increase in the apoptotic percentage in testis from animals under the high-fat diet. Surprisingly, the addition of olive oil to the diet avoided the sub-cellular, efficiency and apoptosis changes observed in hypercholesterolemic rabbits. This paper reports the positive effects of the olive oil addition to the diet in the recovery of testicular efficiency and normal sperm shaping, mechanisms altered by hypercholesterolemia.
Assuntos
Acrossomo/efeitos dos fármacos , Dieta Hiperlipídica/efeitos adversos , Hipercolesterolemia/tratamento farmacológico , Azeite de Oliva/administração & dosagem , Doenças Testiculares/prevenção & controle , Acrossomo/patologia , Animais , Modelos Animais de Doenças , Hipercolesterolemia/induzido quimicamente , Hipercolesterolemia/complicações , Infertilidade Masculina/etiologia , Infertilidade Masculina/prevenção & controle , Masculino , Microdomínios da Membrana/efeitos dos fármacos , Azeite de Oliva/farmacologia , Coelhos , Contagem de Espermatozoides , Espermatogênese/efeitos dos fármacos , Doenças Testiculares/etiologia , TestículoRESUMO
The aim of this study was to evaluate the effect of four methods of sperm selection, on the integrity and stability of the plasma membrane, integrity of the acrosomal membrane and spermatic morphology in frozen/thawed ovine semen. Two types of colloidal silica: colloidal silica-silane and colloidal silica-polyvinylpyrrolidone (PVP), and two aliquots: 1 and 4 ml, were used for sperm selection. Probes FITC-PSA and PI were used to measure the integrity of the plasma and acrosomal membranes. Plasma membrane stability was measured, using fluorescent probes M540 and YOPRO1. Effective reduction in the incidence of spermatozoa with acrosomal pathologies was only achieved using 1 ml colloidal silica-silane. All methods were efficient in select viable and unreacted spermatozoa. Only methods using 1 ml of silica were efficient in decrease spermatozoa stained by PI (death). Methods using silica colloidal-silane were more efficient to decrease apoptotic cells after selection when compared to silica colloidal-PVP. In conclusion, sperm selection in colloidal silica-silane and colloidal silica-PVP improved sperm quality when compared to the controls. The method using 1 ml of colloidal silica-silane is the preferred method because its effectiveness and lower cost.
Assuntos
Membrana Celular/metabolismo , Coloides/química , Criopreservação/métodos , Preservação do Sêmen/métodos , Sêmen/metabolismo , Acrossomo/metabolismo , Acrossomo/patologia , Animais , Sobrevivência Celular , Criopreservação/economia , Criopreservação/veterinária , Citometria de Fluxo , Corantes Fluorescentes , Masculino , Análise do Sêmen/veterinária , Preservação do Sêmen/efeitos adversos , Preservação do Sêmen/economia , Preservação do Sêmen/veterinária , Ovinos , Carneiro Doméstico/fisiologia , Motilidade dos Espermatozoides/fisiologiaRESUMO
Male ICR mice were orally administered samarium nitrate [Sm(NO3)3] to investigate its effects on sperm concentration and sperm quality. After acute exposure to ≥2880.00 mg/kg Sm(NO3)3 via intragastric gavage, sperm motility and acrosome integrity were decreased, and the sperm malformation percentage was increased (P < 0.05). After subchronic exposure to ≥500.00 mg/L Sm(NO3)3 administered via drinking water for 90 days, relative gonad weight, sperm concentration, and sperm quality significantly decreased (P < 0.05). Sperm malformation also increased after subchronic exposure to Sm, which was found to be the most sensitive index. Sperm head malformation accounted for the largest proportion of all types of sperm malformations evaluated. Of the six different subtypes of head malformation, irregular shape accounted for the largest proportion.
Assuntos
Acrossomo/efeitos dos fármacos , Samário/toxicidade , Motilidade dos Espermatozoides/efeitos dos fármacos , Acrossomo/patologia , Animais , Masculino , Camundongos , Camundongos Endogâmicos ICR , Samário/administração & dosagem , Contagem de EspermatozoidesRESUMO
OBJECTIVE: To analyze the seminal plasma proteome and biological functions associated with sperm functional alterations. DESIGN: Cross-sectional study. SETTING: University andrology and research laboratories. PATIENT(S): A total of 156 normozoospermic men. INTERVENTION(S): Sperm mitochondrial activity, acrosome integrity, and DNA fragmentation were evaluated in a semen aliquot. Remaining semen was centrifuged, and seminal plasma was utilized for proteomic analysis (liquid chromatography-tandem mass spectrometry). Patients were divided into percentiles (15%) to form the following groups: substudy 1, high (control, n = 26) and low (study, n = 23) sperm mitochondrial activity; substudy 2, high (control, n = 23) and low (study, n = 22) sperm acrosome integrity; and substudy 3, low (control, n = 22) and high (study, n = 22) sperm DNA fragmentation. Groups were compared using univariate and multivariate analyses. Differentially expressed proteins were used for functional enrichment analysis. MAIN OUTCOME MEASURE(S): Seminal plasma proteome and postgenomic pathways are associated with several sperm functional traits. RESULT(S): In total, 506, 493, and 464 proteins were observed in substudies 1, 2, and 3, respectively. Enriched functions in substudy 1 were intramolecular oxidoreductase activity, aminoglycans catabolism, endopeptidases inhibition, lysosomes, and acute-phase response (study group). In substudy 2, main enriched functions were phospholipase inhibition, arachidonic acid metabolism, exocytosis, regulation of acute inflammation, response to hydrogen peroxide, and lysosomal transport (study group). In substudy 3, enriched functions were prostaglandin biosynthesis and fatty acid binding (study group). We proposed eight, six, and eight seminal biomarkers for substudies 1, 2, and 3, respectively. CONCLUSION(S): Seminal plasma proteome reflects sperm mitochondrial activity reduction, acrosome damage, and DNA fragmentation, with several postgenomic functions related to these alterations.
Assuntos
Sêmen/metabolismo , Proteínas de Plasma Seminal/metabolismo , Espermatozoides/metabolismo , Acrossomo/metabolismo , Acrossomo/patologia , Adulto , Biomarcadores/metabolismo , Cromatografia Líquida , Estudos Transversais , Dano ao DNA , DNA Mitocondrial/metabolismo , Análise Discriminante , Fertilidade , Humanos , Análise dos Mínimos Quadrados , Modelos Lineares , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Análise Multivariada , Proteômica/métodos , Espermatozoides/patologia , Espectrometria de Massas em TandemRESUMO
The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.