Localization and expression of ADAM2 in the dromedary camel testis, epididymis and sperm during rutting season

ADAM2 (fertilin ) is a sperm surface protein reported in several mammalian species. However, the presence of ADAM2 in the male reproductive system and sperm of the camel is not well known. The present study was to clarify the localization and expression of ADAM2 in the dromedary camel testis, epididymis and spermatozoa during rutting season using immunohistochemistry (IHC) and the quantitative real-time polymerase chain reaction (qPCR). Tissue samples were obtained from the testis (proximal and distal) and epididymis (caput, corpus, and cauda) from eight mature male camels. Epididymal and ejaculated sperms were collected from four other fertile camels. IHC analysis clearly showed the localization of ADAM2 protein in the spermatocytes and the round and elongated spermatids of the testis, in the epithelial cells along the epididymis tract, on the posterior head of the sperm within the cauda epididymis, and on the acrosomal cap of both the epididymal and ejaculated sperm. The expression of camel ADAM2 mRNA was significantly higher (P 0.05) in the testis when compared with the epididymis. These findings may suggest an important role of ADAM2 in the fertility of male dromedary camels.(AU)

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) – epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 – the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the αtocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the αtocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.(AU)

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)

Características regionais sobre a ultra-estrutura das células principais do epitélio do ducto epididimal de Agouti paca / Regional ultra structural features of principal cells from the epididymal epithelium lining of Agouti paca

As células principais (P) do epitélio de revestimento do epidídimo de paca foram relacionadas com processos citofisiológicos de endocitoses do tipo adsortiva e de fase fluida, respectivamente, aparentemente realizando também secreção apócrina. Essas funções foram propostas embasando-se em características de ultra-estrutura das células P, em cujos citoplasma observaram-se um expressivo número de vesículas, com diferentes formas, tamanhos e presença de conteúdo internalizado em algumas das vesículas revestidas por endomembranas, ocorrendo ainda caveolas e vesículas diminutas localizadas junto à borda apical de microvilos. Ademais, observaram-se vesículas grandes e revestidas e/ou com superfícies lisas; endossomos, e lisossomos de localização predominantemente apical. Uma via de secreção apócrina foi sugerida com base na ocorrência de expansões (protrusões), citoplasmáticas intraluminais nas células P.(AU)

The principal (P) cells of epididymidis surface epithelium of Agouti paca were related to processes of adsorptive endocytosis and phase-fluid endocytosis, as well as protein secretion apparently also occur.These findings had been proposed on the base the cytoplasmic ultrastructural features of P cells in which were seen an expressive number of vesicles with several shapes, sizes and internalized content occurring also smaller pits and pale small vesicles located next to the apical brush border of microvilli. Moreover, occurred coated vesicles, smooth surface vesicles and great vesicles; multivesicular bodies, endosomes and lysosomes mainly viewed on supranuclear and apical positions. Presence of an appocrine secretory pathway was characterized in P cells through the occurrence of apical cytoplasmic expansions, protruding into the ducts epididymidis luminal compartment.(AU)

Localização histoquímica de algumas hidrolases no epidídimo do gerbilo / Histochemical localization of some hydrolases in the epididymis of the gerbil

Neste estudo verificou-se, principalmente, que a expressão tissular da FoAl esteve presente nas regiões proximais do epidídimo de gerbilo. As reatividades da FoAc e da ATPase foram intensas (fortes), na maioria das regiões do epidídimo caracterizadas, marcadamente ao nível do citoplasma apical das células do epitélio tubular, exceto no corpo epididimal um delgado ístmo de conexão da cabeça à cauda epidiclimárias nesta espécie, com notória hiporeatividade enzimática a praticamente todas as enzimas aqui estudadas. A SDH mostrou também fraca atividade em todas as regiões, e estruturas do ducto epidiclimário do gerbilo, exceto no compartimento intraluminal onde se verificou atividade SDH-intensa nos espermatozóides estocados no lúmen. As reações enzimáticas caracterizadas foram correlacionadas com alguns papéis histofisiológicos tais como a mediação enzimática das fosfatases e ATPase em processos como endocitose, secreção, absorção e processo de transporte ativo das células do epitélio tubular. Uma possível marcação específica de mitocôndrias da peça média dos espermatozóides intraluminais pela SDH foi sugerida, com embasamento em processos metabólicos das mitocôndrias.(AU)

This study mainly showed that alkaline phosphatase expression had been present in the proximal regions of the epididymidis ductus of the gerbil which comprised the initial segment and proximal caput. The reactivities of acid phosphatase and ATPase were strong in the proximal and clistal regions of the epididymidis ductus at the level of the apical cytoplasm and epithelium, except at the corpus level, a very thin isthmus located between the caput and cauda epididymidis, and as a general rule a low enzymatic reactive region of the epididymidis of gerbil. SDH revealed also low activities in ali the regions and regional structures of the duct, except into the luminal content formed by storaged spermatozoa, prior on the cauda leveI. The enzymes presented in the epididymidis were correlated to some histophysiological roles such as the enzymatic mecliation of endocytosis, secretion, absorption and active transport concerning to phosphatases and A TPase and a possible mitochondrial role of SDH could occur at the spermatozoa level in which the middle pieces were formed by a great amount of mitochondria.(AU)

Structural features of the epididymal region of the domestic duck (Anas plathyrynchos) / Características estruturais da região epididimária do pato doméstico (Anas plathyrynchos)

The epididymal region of the domestic duck is composed by efferent ductules, whose histotopology was characterized by the proximal and distal ductules and sequentially by the epididymal duct. The epithelial lining of the efferent ductules was ciliated pseudostratified and formed by columnar cells. Also the ducts epididymidis epithelium was pseudostratified but non-ciliated. Concerning the histomorphometric analysis, the epithelial height mean was significantly greater in the distal efferent ductules, differing from the lower epithelium height mean observed in proximal efferent ductules and epididymal duct. The maximum and minimum diameter mean were significantly greater in the proximal efferent ductules, comparatively to the same diameter means of the other ductules. (AU)

A região epididimária do pato doméstico era composta pelos ductúlos eferentes, os quais hitotopologicamente foram caracterizados como dúctulos eferentes proximal e distal e, sequencialmente, pelo ducto epididimal. O epitélio dos dúctulos eferentes era pseudo-estratificado, formado por células colunares. O epitélio dos ductos epididimários mostrou-se também pseudo-estratificado, mas não ciliado. De acordo com as análises histomorfométricas, a média da altura epitelial foi significativamente maior nos dútulos esferentes distais, diferindo das baixas médias de altura epitelial observadas nos dúctulos eferentes proximais e ducto epididimal. A média dos diâmetros máximos e mínimos foi significativamente maior nos dúctulos eferentes proximais, comparativamente as médias dos mesmos diâmetros dos outros dúctulos. (AU)

Avaliação da biometria testicular, epididimal e de glândulas sexuais acessórias e correlações entre características biométricas e histológicas em carneiros deslanados sprd

O objetivo deste estudo foi avaliar a biometria de partes do trato reprodutor de ovinos deslanados sem padrão racial definido (SPRD) criados no semi-árido nordestino, e correlacionar os dados biométricos testiculares, epididimais e de glândulas sexuais acessórias com variáveis associadas ao processo espermatogênico. Os dados biométricos foram obtidos a partir dos testículos, epidídimos e das glândulas sexuais acessórias de 19 animais abatidos em torno de 60 semanas de idade. O parênquima testicular foi avaliado quanto ao diâmetro, volume e comprimento dos túbulos seminíferos, altura do epitélio germinativo e população de células e Sertoli e germinativas. Foram realizadas análises de correlação de Pearson (p < 0.05) entre as características estudadas e montados modelos de regressão linear simples para estas mesmas variáveis através do programa estatístico StatView, 5.0 (SAS, 1998). Correlações significativas foram verificadas entre todas as medidas testiculares (peso, comprimento, largura e volume) e as outras medidas do trato reprodutor, como comprimento da cauda do epidídimo (CCauEp; r=0,66 a 0,68) e as peso das glândulas sexuais acessórias (r=0,68 a 0,72). Estas também tiveram correlações significativas com o peso do epidídimo e com o CCauEp (r=0,56 e 0,70, respectivamente), o que sugere a existência de uma sinergia no desenvolvimento do trato genital masculino. A largura testicular (LT) foi o parâmetro que apresentou correlações mais significativas com peso, comprimento e volume testiculares (r=0,88; 0,87 e 0,94) e os modelos de regressão que permitem estimar qualquer uma dessas características, a exemplo do volume testicular: VT = -260,17 + 68,479 x LT; R²= 0,95. Esta característica também esteve alta e significativamente correlacionada com importantes parâmetros histológicos, como volume do túbulo seminífero (r=0,96), número de espermátides arredondadas por testículo (r=0,89) e produção diária de espermátides arredondadas (r=0,89). Os modelos de regressão linear simples com coeficientes de determinação (R²) elevados permitem estimar as características histológicas a partir da largura testicular. No presente estudo, foram obtidos, pela primeira vez, os pesos das glândulas vesiculares e bulbo-uretrais em ovinos SPRD e pôde-se observar que animais com maior medidas testiculares também apresentaram elevados parâmetros quantitativos da espermatogênese e maiores epidídimos e glândulas vesiculares. A largura testicular pode ser útil para seleção de reprodutores por ser uma característica de fácil mensuração