RESUMO

Seminal plasma proteins already demonstrated to reflect the testicular environment function and important regulatory mechanisms. However, it is crucial to understand which of these proteins participate in probable altered pathways in testicular germ cell tumours and after unilateral orchiectomy. In this study, we proposed to verify, by a multiplex approach, the levels of DNA damage and apoptosis pathways' proteins, in seminal plasma of men before and after unilateral orchiectomy, and also in control men. Comparing pre- and post-orchiectomy groups, just the apoptosis pathways' proteins presented different levels, in which Bad was lower and Bcl2, Akt, caspase-9, p53 and caspase-8 were higher after orchiectomy. When comparing pre- and post-orchiectomy groups with control, both presented lower levels of ChK1, Chk2, H2AX, p53 and p21, for DNA damage pathway. Regarding the apoptosis pathway, lower levels of JNK, Bcl2, Akt, caspase-9, p53 and caspase-8 and higher levels of Bad were observed before orchiectomy. The post-orchiectomy group did not differ from controls, demonstrating a probable restoration on its proteins levels. We can conclude that testicular tumours can alter both of the assessed pathways, and its removal is associated with a probable restoration of the apoptosis pathway.

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RESUMO

Background: Seminal plasma is a set of secretions from accessory male reproduction organs which infl uence spermaticfunctions. Climate changes that affect gametogenesis may produce low reproduction effi ciency in bulls. Electrophoresisis a great help in the diagnosis of reproduction pathologies and in animal differentiation with regard to fertility within thecontext of climate changes. Current research aims at studying the infl uence of the rainy and dry periods in testicle and epididymis morphometry, semen characteristics and protein of seminal plasma by SDS-PAGE electrophoresis in extensivelybred Nellore and Simental bulls.Materials, Methods & Results: Five Nellore and two Simental bulls, at mean age of four years, were employed. Semencollection was undertaken during the rainy (spring and summer) and dry (autumn and winter) at 15-day intervals. Onehundred and fi fty-four ejaculations were analyzed and scrotal, testicle and epididymis measurement was provided. Samplesof semen plasma were centrifuged and conditioned at -196°C till electrophoresis processing. Proteins were extracted from200 µL from each sample in an extraction buffer composed of 0.625 M Tris - HCl, pH 6.8; 2% SDS, 5% β - mercaptoethanol and 20% glycerol. Proteins were quantifi ed and electrophoreses processed. Data were evaluated by analysis ofvariance and means compared by Tukey´s test at 5%. The expression TV = 0.0396 x (average testicle length) x (scrotalperimeter)2 was used for testicle volume (TV), with TV = 460.14 cm3 for the Simental breed during the rainy season andTV = 571.26 cm3 during the dry season. Nellore bulls showed TV = 524.75 cm3 during the rainy season and 515.13 cm3during the dry season. TV increased in Simental bulls during the dry period, whereas Nellore breed increased during therainy one. There was an increase (P < 0.05) in big and total defects during the rainy season...

Assuntos

RESUMO

Background: Seminal plasma is a set of secretions from accessory male reproduction organs which infl uence spermaticfunctions. Climate changes that affect gametogenesis may produce low reproduction effi ciency in bulls. Electrophoresisis a great help in the diagnosis of reproduction pathologies and in animal differentiation with regard to fertility within thecontext of climate changes. Current research aims at studying the infl uence of the rainy and dry periods in testicle and epididymis morphometry, semen characteristics and protein of seminal plasma by SDS-PAGE electrophoresis in extensivelybred Nellore and Simental bulls.Materials, Methods & Results: Five Nellore and two Simental bulls, at mean age of four years, were employed. Semencollection was undertaken during the rainy (spring and summer) and dry (autumn and winter) at 15-day intervals. Onehundred and fi fty-four ejaculations were analyzed and scrotal, testicle and epididymis measurement was provided. Samplesof semen plasma were centrifuged and conditioned at -196°C till electrophoresis processing. Proteins were extracted from200 µL from each sample in an extraction buffer composed of 0.625 M Tris - HCl, pH 6.8; 2% SDS, 5% β - mercaptoethanol and 20% glycerol. Proteins were quantifi ed and electrophoreses processed. Data were evaluated by analysis ofvariance and means compared by Tukey´s test at 5%. The expression TV = 0.0396 x (average testicle length) x (scrotalperimeter)2 was used for testicle volume (TV), with TV = 460.14 cm3 for the Simental breed during the rainy season andTV = 571.26 cm3 during the dry season. Nellore bulls showed TV = 524.75 cm3 during the rainy season and 515.13 cm3during the dry season. TV increased in Simental bulls during the dry period, whereas Nellore breed increased during therainy one. There was an increase (P < 0.05) in big and total defects during the rainy season...(AU)

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RESUMO

The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.(AU)

Investigou-se a distribuição topográfica da ligação de proteínas seminais à membrana de espermatozoides bovinos epididimários e ejaculados. Utilizando imunocitoquímica e microscopia confocal, avaliaram-se a topografia de ligação das proteínas BSP-A3, albumina, transferrina, prostaglandina D sintetase (PGDS) e nucleobindina 2 (NUC2) à membrana espermática. Os experimentos foram realizados utilizando espermatozoides de cinco touros. Os resultados mostraram que, para espermatozoides epididimários, somente detectou-se a PGDS na crista do acrossomo. Nos espermatozoides ejaculados, a PGDS ligou-se de forma mais intensa à crista acrossômica, enquanto a NUC2 apresentou sinal bastante fraco na peça intermediária e região equatorial. A BSP-A3 ligou-se a todas as regiões estudadas, de forma mais intensa na peça intermediária e acrossomo. Nenhum sinal foi detectado para albumina ou transferrina, seja em espermatozoides epididimários ou ejaculados. Concluiu-se que PGDS, BSP-A3 e NUC2 interagem diretamente com espermatozoides bovinos, e mostrou distribuição topográfica específica. Estes achados permitem melhor compreensão sobre o papel desempenhado por essas proteínas na regulação da função espermática e da fertilidade.(AU)

Assuntos

RESUMO

The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.

Investigou-se a distribuição topográfica da ligação de proteínas seminais à membrana de espermatozoides bovinos epididimários e ejaculados. Utilizando imunocitoquímica e microscopia confocal, avaliaram-se a topografia de ligação das proteínas BSP-A3, albumina, transferrina, prostaglandina D sintetase (PGDS) e nucleobindina 2 (NUC2) à membrana espermática. Os experimentos foram realizados utilizando espermatozoides de cinco touros. Os resultados mostraram que, para espermatozoides epididimários, somente detectou-se a PGDS na crista do acrossomo. Nos espermatozoides ejaculados, a PGDS ligou-se de forma mais intensa à crista acrossômica, enquanto a NUC2 apresentou sinal bastante fraco na peça intermediária e região equatorial. A BSP-A3 ligou-se a todas as regiões estudadas, de forma mais intensa na peça intermediária e acrossomo. Nenhum sinal foi detectado para albumina ou transferrina, seja em espermatozoides epididimários ou ejaculados. Concluiu-se que PGDS, BSP-A3 e NUC2 interagem diretamente com espermatozoides bovinos, e mostrou distribuição topográfica específica. Estes achados permitem melhor compreensão sobre o papel desempenhado por essas proteínas na regulação da função espermática e da fertilidade.

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