Influence of year periods on testicle and epididymal morphometry, semen characteristics and seminal proteins by SDS-PAGE in Zebu and Taurine / Influência das épocas do ano na morfometria testicular e epididimária, características do sêmen e proteínas do sêmen em SDS-PAGE em zebus e taurinos

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)

Influence of year periods on testicle and epididymal morphometry, semen characteristics and seminal proteins by SDS-PAGE in Zebu and Taurine / Influência das épocas do ano na morfometria testicular e epididimária, características do sêmen e proteínas do sêmen em SDS-PAGE em zebus e taurinos

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...

Binding patterns of seminal plasma plasma proteins on bovine epididymal and ejaculated sperm membrane / Topografia de ligação de proteínas do plasma seminal à membrana de espermatozoides bovinos epididimários e ejaculados

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)

Proteoma do plasma seminal de touros da raça Gir - Leiteiro (Bos taurus indicus) clinicamente normais e com hipoplasia testicular

A raça Gir Leiteiro é uma excelente opção para os rebanhos leiteiros no Brasil pelas características apresentadas por seus animais, como rusticidade, adaptabilidade e produção de leite expressiva. A hipoplasia testicular causa problemas reprodutivos nos touros podendo levar à subfertilidade ou, em casos mais graves, à infertilidade. No Brasil, a incidência dessa patologia varia de 5 a 15%. Devido à grande importância que animais da raça Gir Leiteiro têm na pecuária brasileira e aos impactos negativo, tanto em termos de fertilidade quanto economicamente, causados pela hipoplasia testicular, este trabalho objetivou traçar o perfil proteico diferencialmente expresso entre animais com esta patologia e clinicamente normais. Amostras de sêmen de 18 animais (9 hipoplásicos e 9 normais) foram coletadas e o plasma seminal foi centrifugado e submetido à eletroforese bidimensional (2-DE). Os géis foram corados com Coomassie Blue G-250, digitalizados e analisados pelo programa PDQuest 2-D Analysis Software para identificação dos spots. Foram identificados 33 spots diferencialmente expressos entre os dois grupos de animais, sendo 26 mais intensamente expressos em animais hipoplásicos. Os spots foram cortados dos géis e submetidos à espectrometria de massa sendo identificadas 38 proteínas e, dentre estas, o grupo de proteínas conhecidas como Binder of Sperm Proteins (BSP) e a platelet-activating fator acetylhydrolase (PAF AH) foram as mais presentes nos spots identificados nos animais hipoplásicos. Os animais também foram submetidos à avaliação clinico-andrológica e houve diferenças estatísticas quanto ao comprimento, largura e volume testicular, turbilhonamento e defeitos espermáticos (p<0,05).

The Gir - Dairy is an excellent option for dairy herds in Brazil because of the features presented by its animals, such as hardiness, adaptability and expressive milk production. Testicular hypoplasia causes reproductive problems in bulls and can lead to subfertility or, in more severe cases, infertility. In Brazil, the incidence of this disease varies from 5 to 15%. Due to the great importance that Gyr- Dairy animals have in the Brazilian cattle industry and because of the negative impact, not only in economically terms but also in fertility, caused by testicular hypoplasia, this study aimed to determine the differentially expressed protein profile between animals with this disease and clinically normal . Semen samples from 18 animals (9 hypoplastic and 9 normal) were collected and seminal plasma was centrifuged and subjected to two-dimensional electrophoresis (2-DE). Gels were stained with Coomassie Blue G-250, digitized and analyzed using PDQuest program 2-D Analysis Software to identify the "spots". Were identified 33 "spots" differentially expressed between the two groups of animals, 26 more intensely expressed in hypoplastic animals. The "spots" were cut from the gels and subjected to mass spectrometry and identified 38 proteins and, among these, the group of proteins known as Binder of Sperm Proteins (BSP) and platelet-activating factor acetylhydrolase (PAF - AH) were more present in spots identified in hypoplastic animals. The animals also underwent clinical and soundness evaluation and were statistical differences in the length, width and testicular volume, turbulence and sperm defects (p <0.05).