RESUMO
Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm-ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm-egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein-protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved not only in sperm-egg fusion but also in the previous stage of sperm-ZP binding through its specific interaction with human ZP3.
Assuntos
Proteínas do Ovo/genética , Glicoproteínas de Membrana/genética , Receptores de Superfície Celular/genética , Capacitação Espermática/genética , Espermatozoides/metabolismo , Zona Pelúcida/metabolismo , Reação Acrossômica/efeitos dos fármacos , Adulto , Anticorpos/farmacologia , Sítios de Ligação , Ligação Competitiva , Proteínas do Ovo/metabolismo , Proteínas do Ovo/farmacologia , Epididimo/citologia , Epididimo/efeitos dos fármacos , Epididimo/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/metabolismo , Glicoproteínas de Membrana/farmacologia , Ligação Proteica , Receptores de Superfície Celular/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Transdução de Sinais , Capacitação Espermática/efeitos dos fármacos , Espermatozoides/citologia , Espermatozoides/efeitos dos fármacos , Zona Pelúcida/efeitos dos fármacos , Glicoproteínas da Zona PelúcidaRESUMO
In the winter of 2008, cattle on a farm in the province of Neuquen, Argentina died from subacute and chronic liver fluke disease despite four previous treatments with Triclabendazole (TCBZ). In the spring of 2009, a preliminary efficacy test revealed good performance using nitroxynil, whereas TCBZ efficacy was only 18% by egg counts of Fasciola eggs in the faeces. Resistance to Fasciola hepatica to TCBZ has never been reported in South America, so in January of 2010 a controlled trial was conducted to confirm and to define the degree of resistance in this herd. In a clinical trial, the fluke egg output was monitored on Days 14 and 21 and serum enzymes gamma-glutamyl transpeptidase (GGT) and glutamic-oxaloacetic transaminase (GOT) on Days 0 and 21 in 36 calves treated with TCBZ or with closantel. The results showed a reduction of 100% in fluke egg output at Days 14 and 21 for closantel. The mean epg in the TCBZ-treated groups did not decrease. Because of the fact that in this study TCBZ treatment in cattle had no effect, even at double the recommended dose, it is highly indicative that resistance of F. hepatica against TCBZ is present on this farm. The GGT and GOT levels decreased in the closantel-treated group as a result of the treatment at 21 days after dosing. To evaluate the importance of TCBZ resistance in F. hepatica in Argentina, a study on more farms from endemic areas is needed.
Assuntos
Anti-Helmínticos/farmacologia , Benzimidazóis/farmacologia , Doenças dos Bovinos/parasitologia , Fasciola hepatica/crescimento & desenvolvimento , Fasciolíase/veterinária , Hepatopatias/veterinária , Salicilanilidas/farmacologia , Animais , Anti-Helmínticos/uso terapêutico , Argentina , Aspartato Aminotransferases/sangue , Benzimidazóis/uso terapêutico , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/enzimologia , Resistência a Medicamentos , Fasciolíase/tratamento farmacológico , Fasciolíase/enzimologia , Fasciolíase/parasitologia , Fezes/parasitologia , Hepatopatias/tratamento farmacológico , Hepatopatias/enzimologia , Hepatopatias/parasitologia , Contagem de Ovos de Parasitas/veterinária , Salicilanilidas/uso terapêutico , Triclabendazol , gama-Glutamiltransferase/sangueRESUMO
En ésta revisión se discuten las posibles intervenciones para proteger la fertilidad futura de aquellos pacientes oncológicos prepúberes o en la pubertad temprana. En el varón, además de la conocida técnica de crioconservación del semen, se proponen alternativas útiles para niños prepúberes y aquellos adolescentes y jóvenes en los que la enfermedad ha deteriorado significativamente la calidad del semen. Se discute la criopreservación de biopsias testiculares que podrán ser utilizadas para: transplante subcutáneo de tejido, auto y xenotransplante de células germinales y para espermatogénesis in vitro. En la mujer se examina la crioconservación de embriones y ovocitos y la conservación de biopsias de ovario destinadas al auto y xeno transplante y la maduración folicular in vitro. Estadísticas recientes indican que 1 de cada 650 niños sufrirá de alguna forma de cáncer y que aproximadamente el 70 por ciento resultará curado. Sin embargo, éste éxito terapéutico trae aparejada una fuerte incidencia de alteraciones graves de la futura función reproductiva, tanto en hombres como mujeres...
Assuntos
Humanos , Masculino , Feminino , Antineoplásicos/efeitos adversos , Fertilidade , Infertilidade Feminina , Infertilidade Masculina , Ovário , Testículo/transplante , Infertilidade , Infertilidade Feminina , Infertilidade Masculina , Ovário , TestículoRESUMO
En ésta revisión se discuten las posibles intervenciones para proteger la fertilidad futura de aquellos pacientes oncológicos prepúberes o en la pubertad temprana. En el varón, además de la conocida técnica de crioconservación del semen, se proponen alternativas útiles para niños prepúberes y aquellos adolescentes y jóvenes en los que la enfermedad ha deteriorado significativamente la calidad del semen. Se discute la criopreservación de biopsias testiculares que podrán ser utilizadas para: transplante subcutáneo de tejido, auto y xenotransplante de células germinales y para espermatogénesis in vitro. En la mujer se examina la crioconservación de embriones y ovocitos y la conservación de biopsias de ovario destinadas al auto y xeno transplante y la maduración folicular in vitro. Estadísticas recientes indican que 1 de cada 650 niños sufrirá de alguna forma de cáncer y que aproximadamente el 70 por ciento resultará curado. Sin embargo, éste éxito terapéutico trae aparejada una fuerte incidencia de alteraciones graves de la futura función reproductiva, tanto en hombres como mujeres...(AU)