Efficacy of Essential Trace Elements Supplementation on Mineral Composition, Sperm Characteristics, Antioxidant Status, and Genotoxicity in Testis of Tebuconazole-treated Rats / 生物医学与环境科学（英文）

Objective@#This research was performed to evaluate the effect of tebuconazole (TBZ) on reproductive organs of male rats and to assess the protective role of combined essential trace elements in alleviating the detrimental effect of TBZ on male reproductive function.@*Methods@#For this purpose, 48 rats were exposed to 100 mg/kg TBZ, TBZ supplemented with zinc (Zn), selenium (Se), copper (Cu), and iron (Fe), TBZ + (Se + Zn); TBZ + Cu; or TBZ + Fe. The experiment was conducted for 30 consecutive days.@*Results@#TBZ caused a significant perturbation in mineral levels and reduction in reproductive organs weights, plasma testosterone level, and testicular antioxidant enzyme activities. The TBZ-treated group also showed a significant increase in sperm abnormalities (count, motility, and viability percent), plasma follicle-stimulating hormone and luteinizing hormone concentrations, lipid peroxidation, protein oxidation, and severe DNA degradation in comparison with the controls. Histopathologically, TBZ caused testis impairments. Conversely, treatment with trace elements, in combination or alone, improved the reproductive organ weights, sperm characteristics, TBZ-induced toxicity, and histopathological modifications in testis.@*Conclusion@#TBZ exerts significant harmful effects on male reproductive system. The concurrent administration of trace elements reduces testis dysfunction, fertility, and toxicity induced by TBZ.

Impact of testicular sperm extraction and testicular sperm aspiration on gonadal function in an experimental rat model

ABSTRACT Purpose To assess the impact of sperm retrieval on the gonadal function of rats with impaired spermatogenesis by comparing testicular sperm extraction (TESE) to aspiration (TESA). The efficacy of these procedures to sperm obtainment was also compared. Materials and Methods A pilot study showed impaired spermatogenesis, but normal testosterone (T) production after a bilateral orchidopexy applied to 26 rats, which were randomly assigned into four groups: TESE (n=7), TESA (n=7), SHAM (n=6) and Control (n=6). The T levels were measured through comparative analysis after the orchidopexy. Results There was no statistical difference in the animal's baseline T levels after orchidopexy in comparison to the controls: the TESE and TESA groups, 6.66±4.67ng/mL; the SHAM group (orchidopexy only), 4.99±1.96ng/mL; and the Control, 4.75±1.45ng/mL, p=0.27. Accordingly, no difference was found in the postoperative T levels: TESE, 5.35±4.65ng/mL; TESA, 3.96±0.80ng/mL; SHAM, 3.70±1.27ng/mL; p=0.4. The number of sperm cells found through TESE (41.0±7.0) was significantly larger than that found through TESA (21.3±8.1, p=0.001). Moreover, higher tissue weight was found through TESE (0.09±0.02g versus 0.04±0.04g, p=0.04). Conclusions The testicular sperm capture performed in rats through extraction or aspiration, after orchidopexy, did not significantly decrease the T levels. The amount of sperm found through testicular sperm extraction was higher than that through testicular sperm aspiration.

In vitro chemokine (C-C motif) receptor 6-dependent non-inflammatory chemotaxis during spermatogenesis

BACKGROUND: Chemokine (C-C motif) receptor 6 (CCR6) is present in sperm and plays a significant role in sperm motility and chemotaxis acting in the reproductive tracts. However, the expression and functional significance of CCR6 in testis are still poorly understood, especially in the process of spermatogenesis. METHODS AND RESULTS: CCR6 was expressed in spermatogenic cell lines and its expression was shown in an age-dependent upregulation manner from puberty to adulthood in mouse testis. Immunostaining results confirmed the localization of CCR 6 in testis. Further chemotaxis assays demonstrated that spermatogenic cells GC-1 and -2 exhibited a directional movement toward CCR6-specific ligand such as CCL20 or Sertoli cells in vitro. CONCLUSIONS: The present findings indicate that CCR6 is involved in the chemotaxis of spermatogenic cells in vitro and promotes chemotaxis under non-inflammatory conditions during normal spermatogenesis.

Comparison of sexual performance and testicular characteristics of melatonin treated Kivircik and Charollais rams during the non-breeding season / Comparação de desempenho reprodutivo e características testiculares de carneiros Kivircik e Charollais tratados com melatonina durante a estação não reprodutiva

This study was conducted to investigate the effects of melatonin implantation on basic testicular characteristics and reproductive performance of Kivircik and Charollais rams and ewes during the non-breeding season. In this study, 8 Kivircik and 6 Charollais rams were used. Rams were implanted with 54 mg melatonin according to the manufacturer's instructions. At melatonin implantation and at ram introduction the reproductive performance and testicular characteristics were evaluated. Throughout the experimental period, rams were permanently kept outdoors under conditions of natural day length and at ambient temperature. The effects of exogenous melatonin treatments on the reproductive performances of rams and ewes, estrus response, pregnancy rate, litter size and twinning rate of ewes were evaluated in all groups. Libido values were significantly higher in Charollais rams compared to Kivircik rams (P<0.001). Testicular volume (TV) was increased in both ram breeds. Scrotal length (SL) was also increased in both Kivircik and Charollais rams (P<0.01). In conclusion, we showed that the treatment of rams with slow release melatonin implants increased scrotal diameters and testicular volumes in both Kivircik and Charollais rams. Furthermore, melatonin implantation improved the reproductive performances of ewes naturally mated with these melatonin implanted rams during non-breeding season.(AU)

Esse estudo foi conduzido para investigar os efeitos de implantação de melatonina em características testiculares básicas e desempenho reprodutivo de carneiros Kivircik e Charollais durante a estação não reprodutiva. Nesse estudo, oito carneiros Kivircik e 6 Charollais foram utilizados. Foi implantado 54 mg de melatonina nos cordeiros de acordo com as instruções do produtor. Ao implantar a melatonina e à introdução do carneiro, foram avaliados o desempenho reprodutivo e as características testiculares. Ao longo do período experimenta, os cordeiros foram mantidos em ambiente externo sob condições de luz e ritmo circadiano naturais e à temperatura ambiente. O efeito de tratamento com melatonina exógena no desempenho reprodutivo de carneiros e ovelhas, resposta estral, taxa de prenhez, tamanho de ninhada e taxa de gemelaridade foram avaliados em todos os grupos. Valores de libido foram significativamente maiores em Charollais comparados a Kivircik (P<0.001). Volume testicular (VT) estava aumentado em ambas as raças. Comprimento escrotal (CE) também estava aumentado tanto em Kivircik quanto em Charollais (P<0.01). Conclui-se que o tratamento de carneiros com implantes de liberação lenta de melatonina aumentou diâmetro e volume testiculares tanto em carneiros Kivircik quanto em Charollais. Ademais, implantação de melatonina melhorou o desempenho reprodutivo de ovelhas naturalmente cruzadas com esses carneiros implantados com melatonina durante o período não reprodutivo.(AU)

Phytotherapy in reducing glycemic index and testicular oxidative stress resulting from induced diabetes: a review / Fitoterapia na redução da glicemia e do estresse oxidativo testicular resultantes do diabetes experimental: uma revisão

Abstract Oxidative stress plays a main role in the development of diabetes complications. The impairment of gonadal antioxidant potential and endocrine disturbance in diabetic males causes testicular damage and failure in sperm production. Plants have been widely used to control diabetes due their hypoglycemic and antioxidant potential, contributing towards the recovery of testicular function. Current study comprises a review of the literature on the main medicinal plants used in the recovery of testicular oxidative damage in animals with experimental diabetes. Eighteen plant species in the nineteen studies selected from the search strategy were evaluated. Plant extracts were evaluated according to their effects on blood glucose and insulin levels, antioxidant enzymes and oxidant levels, lipid peroxidation, total protein, testosterone levels, gonadosomatic index, diameter of seminiferous tubules, seminiferous epithelium height and integrity, number of germ cells at stage VII and apoptosis in the seminiferous epithelium, sperm production, motility, viability and morphology. After the analysis of the studies, it was observed that plant species, used alone or in combination, may control testicular oxidative damage triggered by diabetes. The antioxidant potential varies among species, with some plants proving to have a better performance in the recovery of reproduction parameters than others.

Resumo O estresse oxidativo desempenha um dos principais papéis no desenvolvimento das complicações do diabetes. Em machos diabéticos, as alterações ao potencial antioxidante gonadal e os distúrbios endócrinos resultam em danos testiculares e falhas na produção de espermatozoides. Plantas têm sido amplamente utilizadas como um método de controle do diabetes devido aos seus potenciais hipoglicemiante e antioxidante, contribuindo para a recuperação da função testicular. Assim, o objetivo deste estudo é fazer uma revisão da literatura sobre as principais plantas medicinais utilizadas na recuperação de danos oxidativos testiculares em animais induzidos ao diabetes experimental. Nos dezenove estudos selecionados a partir da estratégia de busca, dezoito espécies de plantas foram utilizadas. Os extratos das espécies vegetais foram avaliados de acordo com os seus efeitos nas concentrações de glicose e insulina, níveis das enzimas antioxidantes e dos oxidantes, peroxidação lipídica, quantidade de proteínas totais, níveis de testosterona, índice gonadossomático, diâmetro dos túbulos seminíferos, altura e integridade do epitélio seminífero, número de células germinativas no estágio VII, número de células em apoptose no epitélio seminífero, produção, motilidade, viabilidade e morfologia espermática. Depois das análises aos estudos, foi observado que as espécies vegetais, sozinhas ou em combinação, podem ajudar a controlar os danos oxidativos testiculares desencadeados pelo diabetes. Contudo, o potencial antioxidante varia entre as espécies, algumas tenho uma melhor performance na recuperação dos parâmetros reprodutivos do que outras.

Permanent germinal epithelium and reproductive cycle of Atractosteus tropicus (Lepisosteiformes: Lepisosteidae) males, Tabasco, Mexico / Epitelio germinal permanente y ciclo reproductivo en machos de Atractosteus tropicus (Lepisosteiformes: Lepisosteidae) Tabasco, México

Abstract:The tropical gar A. tropicus plays an important ecological role as it regulates other fish stocks in different water bodies in Southeastern México. Nevertheless, wild populations are declining, and one conservation alternative is the aquaculture production and basic knowledge of reproductive biology; for males, this requires the study of germ and somatic structures of testes, to characterize the reproductive cycle, and to provide basic knowledge for exploitation and conservation models and strategies. With this aim, a total of 24 males with an average sL = 47.2 cm were collected from wild populations from the Laguna Pomposú, municipality of Jalpa de Mendez (18°19' - 93°01'12" W), Tabasco, Mexico. Fish were collected with a trawl net and were transported live to the Tropical Aquaculture Laboratory, División Académica de Ciencias Biológicas (DACBiol), Universidad Juárez Autónoma de Tabasco (UJAT). Males were killed by prolonged immersion in MS222. Testes samples were collected from each specimen and were processed using the standard histological procedures, that consisted of dehydration in an ascending ethanol series, xylol, embedding in paraffin, sectioning at 7 µm, and staining with hematoxylin-eosin (HE). The diameter of 20 seminiferous tubules (Dst), height of germinal epithelium (Hge), gonadosomatic index (GSI) and gonad volume (gV) were determined monthly. Based on morphometric and morpho-physiological characteristics, the testes consisted of a network of anastomosed tubules with non-restricted cystic spermatogenesis, and a permanent germinal epithelium. This is the first report of a permanent germinal epithelium in A. tropicus. Five reproductive classes were histologically identified: Class I Regressed; Class II Early Maturation; Class III Mid Maturation; Class IV Late Maturation; Class V Regression. Monthly GSI, gV and Dst values were lower in January and February, the testis showed spermatozoa remains and a regenerating discontinuous germinal epithelium. In March spermiogenesis increased and proliferation of spermatogonia decreased. Male tropical gar followed a seasonal reproductive cycle, indicated by the monthly variation of the reproductive classes and the reproductive season processes observed, and for which temperature and rainfall seem to stimulate reproductive activity and spermiation. Rev. Biol. Trop. 64 (4): 1597-1609. Epub 2016 December 01.

Resumen:A. tropicus tiene un papel ecológico importante, como regulador de otras poblaciones de peces, en los cuerpos de agua de México, pero sus poblaciones silvestres se reducen. Una alternativa de conservación es el cultivo, el cual requiere caracterizar el ciclo reproductivo por medio del estudio de estructuras germinales y somáticas de los testículos, conocimientos que son básicos para formar modelos de aprovechamiento y conservación. Se capturaron mensualmente tres machos sexualmente maduros (N = 24), con un promedio de sL = 47.2 cm en Laguna de Pomposú, Jalpa de Méndez (18°19´59" N - 93°01´12" W), Tabasco, México, de octubre 2009 a septiembre 2010. La técnica de captura fue red de arrastre, se transportaron vivos al laboratorio de acuicultura tropical, DACBiol, UJAT. Los machos recolectados se sacrificaron con baños de inmersión en sobredosis de MS222, los testículos se procesaron para análisis histológico. Se determinó mensualmente el diámetro de 20 túbulos seminíferos (Dst), altura de epitelio germinal (Hge), índice gonadosomático (GSI) y volumen de gónada (gV). Características morfo-fisiológicas del testículo muestran que está constituido de una red de túbulos anastomosados con espermatogénesis quística no restringida, y un epitelio germinal permanente, de nuestro conocimiento es la primera vez que se reporta este tipo de epitelio en Holostei (Lepisosteiformes: Lepisosteidae). Se identificaron cinco clases reproductivas: Clase I Recrudescencia, Clase II Madurez temprana, Clase III Madurez intermedia, Clase IV Maduración tardía, Clase V Regresión, que al contrastarlo con el valor mensual de los indicadores sexuales "GSI, gV, Dst" muestra un patrón de variación; durante enero-febrero se presentan valores bajos, se observa un epitelio germinal discontinuo en regeneración; durante marzo se incrementa la proliferación de espermatogonias disminuyendo la espermatogénesis. Los machos de A. tropicus muestran una actividad reproductora estacional anual, explicado por las variaciones mensuales de los indicadores reproductores, donde la temperatura y la precipitación parecen tener un papel importante como factores que estimulan la actividad reproductora y por tanto la espermiación.

Rediscovering hermaphroditism in Grammatidae with the description of the testicular gland in Brazilian Basslet Gramma brasiliensis / Redescobrindo o hermafroditismo em Grammatidae com a descrição da glândula testicular de Gramma brasiliensis

Abstract Many aspects of sex change in reef fishes have been studied, including behavior and social organization. However, gonad histology remains the most robust way to identify sexual patterns in fishes. Some uncommon tissues remain poorly described, such as the accessory gonadal structures found in species from the Gobiidae family, which are rare in other bony fishes. This is the first report of the testicular gland in Gramma brasiliensis and for the Grammatidae family. Between April 2011 and February 2012 eighty specimens were collected during four dive campaigns on the Taipus de Fora reef (13°56’20”S 38°55’32”W), Bahia, Northeast Brazil, and their sex was determined. Thirteen per cent of the active-females and 90% of the active-males had testicular gland tissue in their ovotestis. This discovery led to additional research into the characteristics of the gland tissue and its relationship with gonadal maturation. Three patterns of testicular gland development were found in Brazilian basslet ovotestis. Both ova and sperm-producing gonad contained testicular gland tissue, and the appearance of this tissue seems to be the first modification of ovotestis tissue marking the beginning of the protogynous sex-change process in G. brasiliensis.

Resumo Diversos aspectos da troca de sexo em peixes recifais vem sendo estudados, incluindo comportamentos e organização social. Entretanto, a histologia das gônadas continua sendo a maneira mais robusta para se identificar padrões sexuais em peixes. Alguns tecidos incomuns, tais como as estruturas anexas a gônada encontradas em espécies da família Gobiidae e raras em outras espécies são pouco estudados. Este trabalho é a primeira descrição da glândula testicular em Gramma brasiliensis e para a família Grammatidae. Entre abril de 2011 e fevereiro de 2012, oitenta espécimes foram coletados durante quatro amostragens no recife de Taipus de Fora (13°56’20”S 38°55’32”W), Bahia, Brasil, e tiveram seus sexos determinados. Treze por cento das fêmeas ativas e noventa por cento dos machos ativos apresentaram tecido da glândula testicular em suas gônadas. Esta descoberta levou ao estudo da características dessa estrutura e sua relação com a maturação gonadal. Foram identificados três padrões de desenvolvimento da glândula testicular nas gônadas do Gramma brasiliensis. Tanto as gônadas produtoras de espermatozoides quanto as de oócitos apresentaram tecido da glândula testicular, e o surgimento desse tecido parece ser a primeira modificação gonadal do início da troca de sexo protogínica em G. brasiliensis.

Influence of scrotal bipartition on spermatogenesis yield and sertoli cell efficiency in sheep / Influência da bipartição escrotal sobre o rendimento da espermatogênese e eficiência das células de Sertoli em ovinos

With the objective to assess the effect of scrotal bipartition on spermatogenesis in sheep, the testes were used from 12 crossbred rams of sheep farms in the municipality of Patos, Paraíba, Brazil, distributed into two groups: GI with six rams with scrotal bipartition, and GII with six rams without scrotal bipartition. The testicular biometry was measured and the testes were collected, fixed in Bouin and fragments were processed to obtain histological slides. The spermatogenesis yield and the Sertoli cell efficiency was estimated by counting the cells of the spermatogenetic line at stage one of the seminiferous epithelium cycle and the Sertoli cells. The results were submitted to analysis of variance with the ASSISTAT v.7.6 program and the mean values were compared by the Student-Newman-Keuls test (SNK) at 5% significance. The testicular biometric parameters did not show statistical difference (p>0.05) between the groups. The meiotic, spermatogenetic and Sertoli cell efficiency were higher in bipartitioned rams (p<0.05), while the mitotic yield did not differ (p>0.05) between GI and GII. The results indicated that there is superiority in the spermatogenetic parameters of bi-partitioned rams, suggesting that these sheep present, as reported in goats, indication of better reproductive indices.

Com o objetivo de avaliar o efeito da bipartição escrotal sobre a espermatogênese em ovinos, foram utilizados os testículos de 12 ovinos sem raça definida oriundos de criadouros do município de Patos-PB, Brasil, distribuídos em dois grupos, GI de seis animais com bipartição escrotal e o GII de seis animais sem bipartição escrotal. Realizou-se a aferição da biometria testicular, em seguida, os testículos foram coletados, fixados em Bouin e fragmentos foram processados para obtenção de lâminas histológicas. Foi estimado o rendimento da espermatogênese e eficiência das células de Sertoli contando-se as células da linhagem espermatogênica no estádio I do Ciclo do Epitélio Seminífero, bem como as células de Sertoli. Os resultados foram submetidos à análise de variância pelo programa ASSISTAT v.7.6 e os valores médios foram comparados pelo teste Student-Newman-Keuls (SNK) a 5% de significância. Os parâmetros de biometria testicular não apresentaram diferença estatística (p>0,05) entre os grupos. Os rendimentos meiótico, espermatogênico e a eficiência das células de Sertoli mostraram-se superiores em animais bipartidos (p<0,05), enquanto o rendimento mitótico não diferiu (p>0,05) entre GI e GII. Os resultados indicaram existir superioridade nos parâmetros espermatogênicos de ovinos bipartidos, sugerindo que estes animais apresentam, assim como constatado em caprinos, indicativo de melhores índices reprodutivos.

Age and education influence the performance of elderly women on the dual-task Timed Up and Go test / Idade e educação influenciam o desempenho de mulheres idosas no teste Timed Up and Go com dupla tarefa

Gait variability is related to functional decline in the elderly. The dual-task Timed Up and Go Test (TUG-DT) reflects the performance in daily activities. Objective To evaluate the differences in time to perform the TUG with and without DT in elderly women with different ages and levels of education and physical activity. Method Ninety-two elderly women perfomed the TUG at usual and fast speeds, with and without motor and cognitive DT. Results Increases in the time to perform the TUG-DT were observed at older ages and lower educational levels, but not at different levels of physical activity. More educated women performed the test faster with and without DT at both speeds. When age was considered, significant differences were found only for the TUG-DT at both speeds. Conclusion Younger women with higher education levels demonstrated better performances on the TUG-DT. .

Alterações da marcha são indícios de declínio funcional em idosos. O TUG com dupla tarefa (TUG-DT) reflete o desempenho das atividades do cotidiano. Objetivo Avaliar as diferenças no tempo de execução do TUG com e sem DT em idosas com diferentes faixas etárias, e níveis de escolaridade e atividade física. Método Noventa e duas idosas foram avaliadas pelo TUG nas velocidades usual e máxima, sem e com DT cognitiva e motora. Resultados Houve aumento no tempo de execução do TUG-DT em idosas com maior faixa etária e menor escolaridade, mas não para diferentes níveis de atividade física. Aquelas com maior escolaridade realizaram o teste mais rápido com e sem DT nas duas velocidades. Com relação à faixa etária, foram obervadas diferenças apenas nos testes com DT nas duas velocidades. Conclusão Idosas mais jovens com maior escolaridade demonstraram um melhor desempenho no TUG com DT. .

The importance of the gubernaculum in testicular migration during the human fetal period

Objectives The objective of this review is to study the role of the gubernaculum in the testicular migration process during the human fetal period. Materials and Methods We performed a descriptive review of the literature about the role of the gubernaculum in testicular migration during the human fetal period. Results In the first phase of testicular migration, the gubernaculum enlarges to hold the testis near the groin and in the second phase the gubernaculum migrates across the pubic region to reach the scrotum. The proximal portion of the gubernaculum is attached to the testis and epididymis and the presence of multiple insertions in the distal gubernaculum is extremely rare. The presence of muscle and nerves in the human gubernaculum is very poor. The gubernaculum of patients with cryptorchidism has more fibrous tissue and less collagen and when the patients are submitted to hormonal treatment, the gubernaculum components alter significantly. Conclusions The gubernaculum presents significant structural modifications during testicular migration in human fetuses. .

Light wavelength dependent circadian and seasonal responses in blackheaded bunting.

Animals in the wild are exposed to daily variations in sun light, viz. duration, intensity and spectrum. Photosensitive blackheaded buntings (Emberiza melanocephala) were exposed to photoperiods differing in the length of light period, wavelengths and intensity. The effects of such light changes were measured on locomotor activity rhythm as well as seasonal responses like development of migratory restlessness: Zugunruhe, body mass and gonadal growth. The results show that the buntings are differentially responsive to light wavelengths and intensities and are indicative of a phase-dependent action of light on the circadian photoperiodic system. These birds seem to use changes in the light variables of the solar environment to regulate their circadian and seasonal responses.

vitro culture of human testicular stem cells on feeder-free condition

Spermatogonial stem cells are subpopulation of spermatogonial cells in testis tissue that support beginning and maintenance of spermatogenesis. Ubiquitin carboxy-terminal hydrolase L1 [UCHL1] could be a specific marker for identification of spermatogonial stem cells including spermatogonial sperm cells [SSCs] in testis tissue and during the culture; therefore we undertook this study to culture these human testicular stem cells [hTSCs] in vitro and approved the presence of human testicular stem cells [hTSCs] by UCHL1, also known as PGP9.5. Enzymatic digestion of human testicular biopsies was done by collagenase IV [4 mg/ml] and trypsin [0.25%]. Differential plating of testicular cells in DMEM/F12 and 10% FBS was applied for 16 hr. Floating cells were collected and transferred onto laminin-coated plates with Stem-Pro 34 media supplemented with growth factors of GDNF, bFGF, EGF and LIF to support self-renewal divisions; testicular stem cell clusters were passaged every 14 days for two months. Spermatogonial cells propagation was studied through Expression of UCHL1 in testis tissue and the entire testicular stem cell culture. Testicular stem cell clusters from 10 patients with obstructive azoospermia were cultured on laminin-coated plates and subsequently propagated for two months. The average of harvested viable cells was approximately 89.6%. UCHL1 was expressed as specific marker in testicular stem cells entire the culture. Human testicular stem cells could be obtained from human testicular tissue by a simple digestion, culturing and propagation method for long-term in vitro conditions. Propagation of these cells approved by specific marker UCHL1, during the culture period

Correlations between testicular hemodynamic and sperm characteristics in rams / Correlações entre a hemodinâmica testicular e as características espermáticas em carneiros

The testicular artery is responsible for the blood supply that reaches the testis and has great importance in heat radiation. Vascular changes in the testis may lead to damage in sperm production, reflected in sperm motility and morphology. The aim of the present study was to evaluate correlations between testicular vascularity and sperm characteristic. Eight adult Santa Ines rams showing different reproductive status were used. The testicular vascularity and sperm characteristics were evaluated fortnightly during 90 days. Color Doppler ultrasonography was used to evaluate the testicular hemodynamic. Resistance index (RI) and pulsatility index (PI) of the testicular artery were evaluated by spectral-Doppler mode. The color-Doppler mode was used to evaluate the blood flow of the pampiniform plexus and testicular parenchyma. The semen analyses assessed were volume, concentration, motility, and morphology. The data were submitted to Pearson´s linear correlations test (p 0.05 was considered significant). No correlations were found between motility and testicular hemodynamic. The percentage of total sperm defects was positively correlated to left and right parenchymal score and to left RI and PI. On the other hand, the pampiniform plexus score was positively correlated to the number of colored pixels and negatively correlated to the RI and PI, for both sides. This study showed that the increase of sperm defect can be related to increase of testicular blood flow; however, more studies are need.

A artéria testicular é responsável pelo fluxo de sangue que chega aos testículos e tem grande importância na termorregulação. Mudanças vasculares nos testículos podem levar à queda da produção espermática, refletindo na motilidade e morfologia. O objetivo deste trabalho foi avaliar as correlações entre a vascularização testicular e as características espermáticas. Foram utilizados oito carneiros adultos Santa Inês com diferentes status reprodutivos. A vascularização testicular e as características seminais foram avaliadas por um período de 90 dias. A ultrassonografia Doppler colorida foi utilizada para avaliar a hemodinâmica testicular. Foram calculados os índices de resistência (RI) e os índices de pulsatilidade (PI) com o modo espectral do Doppler. O modo colorido do Doppler foi utilizado para analisar o fluxo sanguíneo do plexo pampiniforme e do parênquima testicular. As características seminais avaliadas foram o volume, concentração, motilidade e morfologia. Os dados foram submetidos ao teste de correlação linear de Person (P 0.05 foi considerado significativo). Não foram encontradas relações entre motilidade e a hemodinâmica testicular. A porcentagem de defeitos totais correlacionou-se positivamente com o escore de vascularização dos parênquimas direito e esquerdo, e com o RI e PI esquerdos. Também o escore de vascularização dos plexos se correlacionou positivamente com a média de pixels e negativamente com o RI e PI, de ambos os lados. Este trabalho mostrou que o aumento de defeitos espermáticos pode estar correlacionado com o aumento do fluxo sanguíneo nos testículos; contudo, mais estudos são necessários.

Administração de cafeína durante a gestação altera parâmetros testiculares na prole de camundongos C57/BL6 na vida adulta / The testis of the mice C57/BL6 offspring in adulthood have alterations due to maternal caffeine consumption

Metabólitos ativos da cafeína apresentam toxicidade, podendo causar efeitos deletérios em muitos órgãos no período fetal pelo consumo materno. O estudo objetivou avaliar o papel da administração de cafeína durante a gestação em vários parâmetros importantes para a função testicular em camundongos adultos C57BL/6. Fêmeas grávidas foram divididas em: grupo controle (C), que receberam injeções de sol. salina (0,9% NaCl) e grupo cafeína (CF), que receberam diariamente injeções subcutâneas de 20 mg / kg de cafeína. Filhotes tiveram livre acesso à ração padrão desde o desmame até três meses de idade, quando foram mortos. O grupo CF apresentou uma redução no consumo alimentar (C = 4,36 ± 0,14; CF = 3,79 ± 0,05/g, p<0,05) e ganho de massa corporal (C = 25,73 ± 0,14; CF = 21,08 ± 0,41/g, p=0,0001). Ainda este grupo, apresentou alterações estruturais nos testículos da prole, como redução no peso relativo (C = 0,078 ± 0,003; CF = 0,063 ± 0,002/g, p=0,002), diâmetro tubular (C = 455,4 ± 2,7; CF = 393,5 ± 3,1/µm, p=0,0001), lume tubular (C = 310,6 ± 2,5; CF = 254,8 ± 2,9/µm², p=0.0001) e altura do epitélio seminífero (C = 144,8 ± 0,9; CF = 138,7 ± 1,3/µm, p=0,0005) observados pela técnica de morfometria. Testosterona sérica avaliada por quimioluminescência foi reduzida (C = 6,6 ± 2,8; CF = 0,5 ± 0,2/ng/ml, p=0.04). Western Blotting foi utilizado para análise de expressão protéica...

Active metabolites of caffeine present toxicity, may cause deleterious effects on many organs in the fetal period by maternal consumption. This study aimed to evaluate the role of caffeine administration during pregnancy on several parameters of adult male testis in C57BL/6 mice. Pregnant C57BL/6 female mice were divided into two groups (n = 10): Control group (C), dams were injected with the vehicle only (saline 0.9 % NaCl); Caffeine group (CF), dams received daily a subcutaneous injection of 20 mg/kg of caffeine/day (1 mg/mL saline). Pups had free access to standard chow since weaning to 3 months of age, when they were killed. Treatment of dams with caffeine did not produce any visible signs of maternal toxicity. At adulthood CF group presented a reduction in the food consumption (C = 4.36 ± 0.14, CF = 3.79 ± 0.05/g, p<0.05) and body weight gain (C = 25.73 ± 0.14, CF = 21.08 ± 0.41/ g, p=0.0001). Several alterations were seen in testis structure sighted by morphometry: testicular weight (C = 0.078 ± 0.003, CF = 0.063 ± 0.002/g, p=0.002), tubular diameter (C = 455.4 ± 2.7, CF = 393.5 ± 3.1/µm, p=0.0001), tubular lumen (C = 310.6 ± 2.5, CF = 254.8 ± 2.9/µm², p=0.0001) and epithelial height (C = 144.8 ± 0.9; CF = 138.7 ± 1.3/µm, p=0.0005). Chemioluminesense was performed to evaluate testosterone serum levels, which was decreased (C = 6.6 ± 2.8, CF = 0.5 ± 0.2/ng/ml, p=0.04). Western blotting showed an increase in the protein expression of leptin’s receptor...

Reproducción Estacional en el Macho / Seasonal Reproduction in the Male

Hay una sorprendente relación entre el ambiente y adaptaciones de la conducta reproductiva, muy evidente en los reproductores estacionales que pueden reproducirse en días cortos o largos, de acuerdo a factores proximales, especialmente el fotoperiodo luminoso que provoca cambios fotoneuroendocrinos. Estos, involucran fotoreceptores, un reloj biológico y el aparato neuroendocrino. Las gonadotrofinas (GT), el desarrollo gonadal, la retroalimentación negativa de las GT por los esteroides sexuales, la intervención de las fibras retino-hipotalámico y los núcleos supraquiasmáticos así como la secreción de melatonina, intervienen en esta regulación. El pulso generador del hipotálamo (eminencia media) es importante en el control de la adenohipófisis respecto de la secreción de LH y FSH. En el testículo, las endocrinocitos intersticiales (de Leydig) (que secretan testosterona y también estrógenos), establecen un asa de retroalimentación con la adenohipófisis y el hipotálamo en un circuito de asa larga, corta y ultracorta, donde neuronas neuroendocrinas tienen un rol importante. Los sustentocitos (células de Sertoli) (intratubulares) son importantes por su rol mecánico, trófico y metabólico respecto a las células germinales y la secreción de activina e inhibina, que provoca o inhibe la secreción de FSH respectivamente. Los sustentocitos también secretan muchas proteínas específicas entre las cuales se encuentra la proteína que liga andrógenos (ABP), importante porque concentra 100 veces la testosterona en el parénquima testicular. La secreción tónica, por pulsos de GT, especialmente LH, es debida a actividad hipotalámica a través del control de generación de estos pulsos que se inicia en la pubertad. La reproducción en el potro y el toro se presentan como ejemplos.

There is a surprising interrelationship between environments and adaptation of reproductive behaviour, very evident in seasonal breeders; which may reproduce in long or short days, according to proximal factors, mainly the light photoperiod which triggers photoneuroendocrine changes. These involve photoreceptors, a clock and the neuroendocrine apparatus. Gonadotropins (GT), gonadal development, negative feed back of GT done by sexual steroid, the intervention of the retino-hypothalamic fibers, and suprachiasmatics nucleus as well as melatonine secretion, intervene in this regulation. Of importance is the pulse generator of the hypothalamus (medial eminence) and its control of adenohypofisis for the secretion of LH and FSH. In the testis interstitial endocrine cells (Leydig)(secreting testosterone and also estrogens), establish a feed back loop with the adenohypofisis and hypothalamus in a circuit of long, short and ultra short circuit with neuroendocrine neurons playing a key role. Sustentocyte intratubular (Sertoli)are also important for their trophic, mechanic and metabolic relationships with the germ cells, and the secretions of activine and inhibine, which triggers or inhibits FSH secretions respectively. Sustentocyte also secrete many specifics proteins among which ABP (Androgen Binding Protein) is important because it concentrates 100 fold testosterone in the testicular parenchyme. Tonic secretion by pulses of GT, mainly LH, is due to hypothalamic activity with the control generation of these pulses by puberty. Reproduction in the stallion and the bull are presented as examples.

Avaliação da morfologia testicular e dos parâmetros espermáticos em ratos espontaneamente hipertensos e tratados com enalapril / Evaluation of testicular morphology and sperm parameters in spontaneously hypertensive rats treated with enalapril

A hipertensão arterial sistêmica (HAS) é um problema de saúde pública, geralmente associada a outras doenças, como obesidade, diabetes, doença renal, aterosclerose, acidente vascular cerebral (AVC) e identificado como um dos fatores de risco mais prevalentes para o desenvolvimento de doenças cardiovasculares. Orgãos-alvo, como coração, rins, cérebro e olhos, são comumente afetados em pacientes hipertensos. No entanto, o dano testicular causado pela hipertensão não foi claramente definido. A hipertensão é um fator de risco bem estabelecido para a disfunção erétil, mas sua relação com o dano testicular e a fertilidade masculina não é claramente compreendida. Este estudo avalia a morfologia testicular e alguns parâmetros espermáticos de ratos espontaneamente hipertensos (SHR), virgem de tratamento e tratados com enalapril. Ratos SHR foram distribuídos em dois grupos, um grupo hipertenso (H), e um grupo tratado com enalapril (HE). Ratos Wistar-Kyoto (WKY) foram utilizados como controles. A pressão arterial sistólica foi medida semanalmente, até o final do experimento. A concentração de espermatozóides, motilidade e viabilidade foram determinadas em amostras coletadas da cauda do epidídimo. Métodos estereológicos foram usados para analisar objetivamente a morfologia testicular macroscopicamente e microscopicamente. Todos os dados foram analisados por ANOVA com pós-teste de Tukey, considerando p <0,05. Ao final do experimento a pressão arterial sistólica no grupo HE (153,9 mmHg ± 21,03 ) foi semelhante a dos animais pertencentes ao grupo WKY (153,4 ± 24,41) e menor que a dos animais H (205,1 ± 24,9). A concentração espermática do grupo H (1,31 x 107 sptz/ml ± 0,27) foi inferior à do grupo WKY (2,11 x 107 sptz/ml ± 0,34), entretanto o controle da pressão arterial com o enalapril melhorou este parâmetro e a concentração espermática do grupo HE (2,46 x 107 sptz/ml ± 0,54) foi semelhante a do WKY...

Hypertension is a major public health problem, usually associated with other disorders such as obesity, diabetes, kidney disease, atherosclerosis, stroke and identified as one of the most prevalent risk factors for developing cardiovascular diseases. Target organs, such as heart, kidney, brain and eyes, are very commonly affected in hypertensive patients. However the testicular damage caused by hypertension has not been clearly defined. Hypertension is a well-established risk factor for erectile dysfunction, but its relation to testicular damage and male fertility is not clearly understood. This study evaluates the testicular morphology and some spermatozoid parameters of spontaneously hypertensive rats (SHR) untreated and treated with enalapril. SHR rats were assigned into two groups, a hypertensive group (H), and an enalapril treated group (HE). Wistar-Kyoto rats (WKY) were used as controls. Systolic blood pressure was measured weekly until at the end of the experiment. The spermatozoid concentration, motility and viability were determined in epididimal tail collected sample. Stereological methods were used to analyze testicular morphology macroscopically and microscopically. Data were analyzed by one-way ANOVA and Tukey´s post test, considering p<0.05. At the end of the experiment systolic blood pressure in the HE group (153,9 mmHg ± 21,03 ) was similar to WKY animals (153,4 ± 24,41), and lower than H animals (205,1 ± 24,9). Sperm concentration of the H group (1,31 x 107 sptz/ml ± 0,27) was lower than WKY group (2,11 x 107 sptz/ml ± 0,34). The blood pressure control with enalapril improved this parameter and HE group (2,46 x 107 sptz/ml ± 0,54) was similar to WKY...

New perspectives in the diagnosis of pediatric male hypogonadism: the importance of AMH as a Sertoli cell marker / Novas perspectivas no diagnóstico do hipogonadismo pediátrico masculino: a importância do AMH como marcador de células de Sertoli

Sertoli cells are the most active cell population in the testis during infancy and childhood. In these periods of life, hypogonadism can only be evidenced without stimulation tests, if Sertoli cell function is assessed. AMH is a useful marker of prepubertal Sertoli cell activity and number. Serum AMH is high from fetal life until mid-puberty. Testicular AMH production increases in response to FSH and is potently inhibited by androgens. Serum AMH is undetectable in anorchidic patients. In primary or central hypogonadism affecting the whole gonad and established in fetal life or childhood, serum AMH is low. Conversely, when hypogonadism affects only Leydig cells (e.g. LHβ mutations, LH/CG receptor or steroidogenic enzyme defects), serum AMH is normal or high. In pubertal males with central hypogonadism, AMH is low for Tanner stage (reflecting lack of FSH stimulus), but high for the age (indicating lack of testosterone inhibitory effect). Treatment with FSH provokes an increase in serum AMH, whereas hCG administration increases testosterone levels, which downregulate AMH. In conclusion, assessment of serum AMH is helpful to evaluate gonadal function, without the need for stimulation tests, and guides etiological diagnosis of pediatric male hypogonadism. Furthermore, serum AMH is an excellent marker of FSH and androgen action on the testis.

As células de Sertoli são a população de células mais ativa nos testículos durante a primeira e segunda infância. Neste período, o hipogonadismo só pode ser evidenciado sem o uso de testes estimulatórios se a função das células de Sertoli for avaliada. O AMH é um marcador útil do número e da atividade das células de Sertoli no período pré-puberal. A concentração sérica de AMH é alta da metade da vida fetal até a metade da puberdade. A produção de AMH pelos testículos aumenta em resposta ao FSH e é potencialmente inibida por androgênios. O AMH sérico não é detectável em pacientes anorquídicos. No hipogonadismo central ou primário afetando a gônada inteira, ou estabelecido na vida fetal ou infância, a concentração de AMH sérica é baixa. Por outro lado, quando o hipogonadismo afeta apenas as células de Leydig (por exemplo, nas mutações, LHβ, defeitos do receptor de LH/CG ou das enzimas esteroidogênicas), a concentração de AMH sérico é normal ou alta. Em meninos púberes com hipogonadismo central, a concentração de AMH é baixa para o estágio na escala de Tanner (refletindo a falta de estímulo pelo FSH), mas alta para a idade (indicando a falta do efeito inibidor da testosterona). O tratamento com FSH provoca um aumento do AMH sérico, enquanto a administração de hCG aumenta os níveis de testosterona, que fazem a downregulation do AMH. Em conclusão, a concentração sérica de AMH é útil na avaliação da função gonadal, excluindo a necessidade de testes estimulatórios, e direciona o diagnóstico etiológico do hipogonadismo pediátrico masculino. Além disso, o AMH sérico é um marcador excelente da ação do FSH e dos androgênios nos testículos.

Estructura testicular y calidad seminal en ratones seleccionados por peso / Testicular structure and semen quality in mice lines selected for weight

La capacidad reproductiva femenina se modificó como consecuencia de la selección de peso realizada en un par de líneas de selección divergente durante 50 generaciones (s: bajo peso; s': alto peso), originadas a partir de una población testigo (t) de ratones CF1 y criadas en la Facultad de Ciencias Veterinarias (UNR). Se probó la existencia de diferencias reproductivas atribuibles al genotipo paterno observadas en el tamaño de camada al nacimiento y en los días transcurridos desde el ingreso a servicio hasta la parición. Con el propósito de explicar las diferencias se comparó la estructura testicular - peso testicular (PTEST), diámetro del túbulo seminífero (DTS), altura de su epitelio (AES) - y la calidad seminal - número de espermatozoides (NE), porcentaje de células anormales (AN), motilidad y desplazamiento de las células espermáticas (Parámetros CASA) -, en machos adultos de 10 a 14 semanas de edad. Se estimó el promedio del DTS y AES en 100 túbulos seminíferos por individuo. El semen se obtuvo de la porción caudal de los epidídimos y de los conductos deferentes. El recuento de espermatozoides se realizó en cámara de Neubauer hemocitométrica y el porcentaje de células anormales se calculó sobre 100 espermatozoides por extendido. Los parámetros de motilidad se determinaron con analizador computarizado de semen Ceros 12.1. La selección por peso diferenció significativamente a las líneas en el mismo sentido de la selección practicada para PTEST, AES y NE. Los machos de la línea más pesada (s') presentaron PTEST y AES mayores y tuvieron un mayor número de espermatozoides al compararlos con los de las líneas s y t (p<0,05). La calidad del semen, bajo las condiciones del setup utilizado, fue similar para los machos de las tres líneas. Por lo tanto, la calidad del semen no aportó a la explicación de las diferencias de fertilidad masculina entre las líneas.

Reproductive female capacity was affected by the genetic selection performed in a pair of lines of two-way selection of body weight for 50 generations (s: downward selection; s': upward selection). These lines were originated from a control CF1 mice population (t) at the Faculty of Veterinary Sciences (UNR). The existence of reproductive differences attributed to the male genotype in the litter size and in the interval from the service beginning to first parturition was proven. With the aim of explaining them, the testicular structure - testicular weight (PTEST), vas deferens diameter (DTS), epithelium height (AES) - and the semen quality - spermatozoa number (NE), abnormal cell percentage (AN), motility and movement of spermatic cells (CASA parameters) were compared, in adult males between 10 and 14 weeks of age. The means of the DTS and AES were estimated for 100 seminiferous tubules per individual. Semen was obtained from the caudal portion of the epididymides and vasa deferentia. Sperm count was performed in Neubauer hemocytometer and abnormal cell percentage was estimated over 100 spermatozoa per sample. Motility parameters were determined with a Ceros 12.1 semen computerized analyzer. Body weight selection occasioned significant differences between lines in the same way of the selection performed for PTEST, AES and NE. Males of the heaviest line (s') show heavier testicles, higher seminiferous epithelium and, consequently, a higher number of spermatozoa when compared with s and t males (p<0,05). Semen quality under the employed setup conditions was similar for males of the three lines. Therefore, semen quality does not contribute to the explanation of the fertility differences between lines.

Morfología testicular del yaque Leiarius marmoratus (Pisces: Siluridae) en estadio de madurez reproductiva / Testicular morphology of yaque Leiarius marmoratus(Pisces: Siluridae) in maturity reproductive stage

El conocimiento de la morfología y fisiología del testículo, permite generar bases para el entendimiento del comportamiento reproductivo de las especies, ya que diferencias anatómicas están relacionadas con el desempeño reproductivo. Con el fin de contribuir al conocimiento biológico y reproductivo del yaque Leiarius marmoratus, fueron analizadas las características anatómicas, morfológicas y funcionales del testículo en animales durante el estado de madurez reproductiva. En L. marmoratus los testículos están ubicados ventralmente a la vejiga gaseosa y presentan numerosas y largas digitaciones que terminan en un conducto espermático. Microscópicamente se encontró que el testículo es espermatogonial irrestricto de tipo tubular, donde la porción anterior presenta actividad espermatogénica, que disminuye en la porción media y desaparece en la región distal, dando paso a tejido glandular con actividad secretora, que actúa como vesícula seminal. Al interior de cada túbulo testicular, se observan cistos que contienen células espermáticas en el mismo estadio de desarrollo. Basados en las características microscópicas, fueron identificados espermatocitos, espermátides y gran cantidad de espermatozoides libres en el lumen. También fueron identificadas células glandulares, tejido muscular liso y tejido epitelial. En el ducto espermático (región media y distal), se encontró abundante secreción acidofílica que en algunas regiones estaba acompañada de espermatozoides libres.

Knowledge of the morphology and physiology of the testis, leads to generate bases for understanding the reproductive behavior of species, because the anatomical differences are related to reproductive performance. To contribute to biologic and reproductive knowledge of Yaque Leiarius marmoratus, anatomic, morphologic and functional characteristics of the testis in animals in a state of reproductive maturity were analyzed. In L. marmoratus, the testicles are located ventrally to the gas bladder and have many long finger-like structures that finish in spermatic duct. Microscopically it was found that the testis is espermatogonial unrestricted and tubular type, where the anterior portion presented spermatogenic activity, which diminishes in the medium portion and disappears in the caudal portion, leading to tissue with glandular secretory activity serving as a seminal vesicle. Inside each tubule of the testis, there are cysts containing sperm cells within the same stage of development. Based on microscopic features, were identified spermatocytes, spermatids and a large amount of free sperm into the lumen. Glandular cells were also identified, as well as smooth muscle tissue and epithelial tissue. In the spermatic duct (medium and distal portion), acidophilous secretion was abundant and in some regions were accompanied by free spermatozoa.

Sperm morphological characteristic and mating behaviour of Proviron® treated West African dwarf bucks with testicular degeneration / Características morfológicas de los espermatozoides y comportamiento en el apareamiento de la cabra enana del Oeste Africano con degeneración testicular tratadas con Proviron®

This study was conducted to determine the effect of scrotal insulation and post insulation Proviron® treatment on sperm morphological characteristics and mating behavior. Twelve healthy WAD bucks, free from any clinical or andrological disorder were used. Scrotal insulation was done using insulating bags for 30 days. After insulation, the bucks were divided into two groups. The treatment group received 100mg/head /week of Proviron® for 3 weeks. Semen collection was done using the electroejaculation method in all phases of the study. The proviron® treated bucks, when compared with insulation and post insulation untreated phases, showed significant reduction (p<0.05) in sperm cell abnormalities which reduced from 15.89 +/- 22.89 in post insulation untreated phase to 2.81 +/- 0.83 in post insulation treated phase. The Proviron® treated bucks also showed increased physical vigor by riding, mounting and fighting their untreated counterparts.

Este estudio se realizó para determinar el efecto de aislamiento escrotal y aislamiento posterior al tratamiento con Proviron® sobre las características morfológicas de los espermatozoides y el comportamiento del apareamiento. Fueron utilizadas 12 cabras enanas del Oeste Africano, sanas, libres de cualquier desorden clínico andrológico. El aislamiento escrotal se realizó utilizando bolsas de aislamiento durante 30 días. Después de aislamiento, los machos se dividieron en dos grupos. El grupo de tratamiento recibió 100mg/cada una por semana de Proviron® durante 3 semanas. La recolección de semen se realizó mediante el método de electroeyaculación en todas las fases del estudio. Las cabras tratadas con Proviron®, cuando se compararon con las fases post-aislamiento no tratadas, mostraron una reducción significativa (p<0,05) en las anormalidades de células espermáticas, las cuales se redujeron desde 15,89 +/- 22,89 en las fases post-aislamiento sin tratamiento a 2,81 +/- 0,83 en las fases post-aislamiento tratadas. Las cabras tratadas con Proviron® también mostraron un incremento en el vigor físico, la monta y la lucha contra sus homólogos no tratados.