RESUMO
Rat epididymal protein DE associates with the sperm surface during maturation and participates in sperm-egg fusion. Immunization of male rats with DE raised specific antibodies and produced a significant reduction in the animals' fertility. The present study focused on determining the in vivo mechanism involved in fertility inhibition. Wistar males were injected with DE, and antibody levels and animal fertility were evaluated. Results revealed an association between the two parameters, since animals with absorbance values lower than 0.5 in ELISA presented high fertility rates (66%, 100%) while those with absorbance values higher than 0.5 exhibited the lowest fertility rates (0%, 33%). Histological studies showed no evidence of orchitis, epididymitis, or vasitis in DE-immunized animals. ELISA results revealed the presence of anti-DE antibodies in epididymal and vas deferential fluids. Indirect immunofluorescence and ELISA experiments indicated that these antibodies would not interfere with the synthesis or secretion of DE or with its association with the sperm surface. Finally, while epididymal sperm recovered from DE-immunized animals presented no changes in motility, viability, or ability to undergo capacitation and acrosome reaction, they exhibited a significant decrease in their ability to fuse with zona-free eggs, with no effect on their ability to bind to the oolemma. Together these results indicate that immunization of male rats with epididymal protein DE specifically interferes with the sperm fertilizing ability, supporting the use of epididymal proteins for contraceptive vaccine development.
Assuntos
Anticoncepção Imunológica , Imunização , Metaloproteínas/imunologia , Interações Espermatozoide-Óvulo , Hormônios Testiculares/imunologia , Animais , Anticorpos/imunologia , Anticorpos/farmacologia , Proteínas Secretadas pelo Epidídimo , Epididimo/imunologia , Feminino , Imunofluorescência , Masculino , Ratos , Ratos Wistar , Capacitação Espermática , Motilidade dos Espermatozoides , Espermatozoides/fisiologiaRESUMO
A commercially available staining kit (Spermac) was combined with swelling in a hypoosmotic medium (HOS) for simultaneous assessment of viability and acrosome reaction in mouse spermatozoa. We compared the results obtained with the combined technique (HOS-Spermac) with those obtained with currently used techniques: the chlortetracycline fluorescence assay and eosin exclusion. The results obtained with HOS-Spermac were the same as those obtained with the chlortetracycline fluorescence assay. Viability assessment with HOS-Spermac showed a good correlation with the percentage of spermatozoa showing eosin dye exclusion. Using this novel technique, we studied the effect of a nitric oxide synthase inhibitor (NG-nitro-L-arginine methyl ester, L-NAME) on the acrosome reaction. L-NAME produced a dose-dependent inhibition of spontaneous acrosome reaction and its inhibitory effect was specifically counteracted by L-arginine. We conclude that HOS-Spermac provides a simple and reliable tool for assessment of the acrosome reaction and that nitric oxide synthase participates in this important function of the spermatozoon.
Assuntos
Acrossomo/enzimologia , Óxido Nítrico Sintase/antagonistas & inibidores , Espermatozoides/enzimologia , Acrossomo/efeitos dos fármacos , Animais , Arginina/farmacologia , Tamanho Celular , Sobrevivência Celular/fisiologia , Inibidores Enzimáticos/farmacologia , Corantes Fluorescentes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase/metabolismo , Espermatozoides/citologia , Espermatozoides/ultraestrutura , Coloração e Rotulagem/métodos , Fatores de TempoRESUMO
Se presenta una serie de 64 pacientes en quienes se hizo cirugía de las suprarrenales. La edad promedio fue 45 años, con similar distribución para ambos sexos. La patología más frecuente fue feocromocitoma 33%, luego el hiperaldosteronismo primario 13% y el hipercortisolismo 17%. Carcinomas no funcionantes 6% y causas misceláneas en 8%. Se hizo autotrasplante de médula suprarrenal al encéfalo por enf. de Parkinson en 15 (23%) pacientes. Se discuten los fundamentos diagnósticos, bioquímicos y métodos de imágenes. Se presentan los accesos quirúrgicos y resultados. La morbilidad baja y ausencia de mortalidad estimulan esta cirugía como un tratamiento electivo de diversas patologías adrenales