Intertubular morphometric and ultrastructural testes analyses in mdx mice

Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.(AU)

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.(AU)

Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells

This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3β-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.

Estágio reprodutivo, histologia e morfometria sazonal do testículo de Dermanura cinerea (Chiroptera: Phyllostomidae) em fragmento de Mata Atlântica no Litoral Sul de Pernambuco, Brasil / Reproductive stage, histology and seasonal morphometry of the testes of Dermanura cinerea (Chiroptera: Phyllostomidae) in Atlantic Forest fragment on the South Coast of Pernambuco, Brazil

This study analyzed the reproductive stage, histology and morphometry of the testes of Dermanura cinerea (Gervais, 1856) in Atlantic Forest fragment on the South Coast of Pernambuco. Specimens were captured by mist net, and collections occurred monthly over two consecutive nights for 18 months. The meteorological data were grouped in rainy and dry months. Eighteen adult specimens were used, which were classified in males with descendant testes and with testes no descendant. For the histological analysis, the testes were collected, fixed and processed following the routine histological technique. The slides obtained were stained by Hematoxylin-Eosin. In the morphometric analysis the area of occupation of the tubular and intertubular compartment were measured, the number of Leydig, Sertoli, spermatocytes and elongated spermatids were quantified. The morphometric data were submitted to statistical analysis. Dermanura cinerea presented higher sperm production activity in dry months and higher hormone production in rainy months. This information is also related to the reproductive pattern of the females and to the increase in rainfall indexes, since it is a determinant factor in the mediation of the food availability of the frugivorous species, such as D. cinerea in the Atlantic Forest area of Pernambuco.(AU)

Esse estudo analisou o estágio reprodutivo, a histologia e a morfometria dos testículos de Dermanura cinerea (Gervais, 1856) em fragmento de Mata Atlântica no Litoral Sul de Pernambuco. Os espécimes foram capturados por redes de neblina e as coletas ocorreram mensalmente, ao longo de duas noites consecutivas, durante 18 meses. Os dados meteorológicos foram agrupados em meses chuvosos e secos. Foram utilizados 18 espécimes adultos, os quais foram classificados em machos com testículos descendentes e não descendentes. Para as análises histológicas, os testículos foram coletados, fixados e processados seguindo a técnica histológica de rotina. As lâminas obtidas foram coradas por Hematoxilina-Eosina. Nas análises morfométricas foram mensuradas a área de ocupação do compartimento tubular e intertubular, quantificados o número de células de Leydig, de Sertoli, de espermatócitos e de espermátides alongadas. Os dados morfométricos foram submetidos às análises estatísticas. Dermanura cinerea apresentou maior atividade de produção espermática em meses secos e maior produção hormonal em meses chuvosos. Essas informações estão relacionadas também com o padrão reprodutivo das fêmeas e com a elevação nos índices pluviométricos, já que é um fator determinante na mediação da disponibilidade alimentar das espécies frugívoras, como D. cinerea em área de Mata Atlântica de Pernambuco.(AU)

Estágio reprodutivo, histologia e morfometria sazonal do testículo de Dermanura cinerea (Chiroptera: Phyllostomidae) em fragmento de Mata Atlântica no Litoral Sul de Pernambuco, Brasil / Reproductive stage, histology and seasonal morphometry of the testes of Dermanura cinerea (Chiroptera: Phyllostomidae) in Atlantic Forest fragment on the South Coast of Pernambuco, Brazil

This study analyzed the reproductive stage, histology and morphometry of the testes of Dermanura cinerea (Gervais, 1856) in Atlantic Forest fragment on the South Coast of Pernambuco. Specimens were captured by mist net, and collections occurred monthly over two consecutive nights for 18 months. The meteorological data were grouped in rainy and dry months. Eighteen adult specimens were used, which were classified in males with descendant testes and with testes no descendant. For the histological analysis, the testes were collected, fixed and processed following the routine histological technique. The slides obtained were stained by Hematoxylin-Eosin. In the morphometric analysis the area of occupation of the tubular and intertubular compartment were measured, the number of Leydig, Sertoli, spermatocytes and elongated spermatids were quantified. The morphometric data were submitted to statistical analysis. Dermanura cinerea presented higher sperm production activity in dry months and higher hormone production in rainy months. This information is also related to the reproductive pattern of the females and to the increase in rainfall indexes, since it is a determinant factor in the mediation of the food availability of the frugivorous species, such as D. cinerea in the Atlantic Forest area of Pernambuco.

Esse estudo analisou o estágio reprodutivo, a histologia e a morfometria dos testículos de Dermanura cinerea (Gervais, 1856) em fragmento de Mata Atlântica no Litoral Sul de Pernambuco. Os espécimes foram capturados por redes de neblina e as coletas ocorreram mensalmente, ao longo de duas noites consecutivas, durante 18 meses. Os dados meteorológicos foram agrupados em meses chuvosos e secos. Foram utilizados 18 espécimes adultos, os quais foram classificados em machos com testículos descendentes e não descendentes. Para as análises histológicas, os testículos foram coletados, fixados e processados seguindo a técnica histológica de rotina. As lâminas obtidas foram coradas por Hematoxilina-Eosina. Nas análises morfométricas foram mensuradas a área de ocupação do compartimento tubular e intertubular, quantificados o número de células de Leydig, de Sertoli, de espermatócitos e de espermátides alongadas. Os dados morfométricos foram submetidos às análises estatísticas. Dermanura cinerea apresentou maior atividade de produção espermática em meses secos e maior produção hormonal em meses chuvosos. Essas informações estão relacionadas também com o padrão reprodutivo das fêmeas e com a elevação nos índices pluviométricos, já que é um fator determinante na mediação da disponibilidade alimentar das espécies frugívoras, como D. cinerea em área de Mata Atlântica de Pernambuco.

Numeric and volumetric changes in Leydig cells during aging of rats

Purpose: To analyze the effects of aging in rats on the nuclear volume, cytoplasmic volume, and total volume of Leydig cells, as well as their number. Methods: Seventy-two Wistar rats were divided into six subgroups of 12 rats, which underwent right orchiectomy at 3, 6, 9, 12, 18, and 24 months of age. The weight and volume of the resected testicles were assessed. A stereological study of Leydig cells was conducted, which included measurements of cell number and nuclear, cytoplasmic, and total cell volumes. Results: The weight and volume of the resected testicles showed reductions with age. Only the subgroup composed of 24-month old rats showed a decrease in the nuclear volume of Leydig cells. Significant reductions in the cytoplasmic volume and total volume of Leydig cells were observed in 18- and 24-month old rats. The number of Leydig cells did not vary significantly with age. Conclusions: Aging in rats resulted in reduction of the nuclear, cytoplasmic, and total cell volumes of Leydig cells. There was no change in the total number of these cells during aging.(AU)

Histometria dos túbulos seminíferos em testículos de ovinos Santa Inês e mestiços Santa Inês e Dorper / Histometry of seminiferous tubules in testes of ovine Santa Ines and mongrel Santa Inês and Dorper

The morphometric measurements of the testicles is intended to measure the dimensions of constituenttestis parenchyma quantitatively evaluating spermatogenesis. This study aimed to analyze the spermatogenesisof sheep Santa Ines and crossbred Santa Ines and Dorper watching the racial crossing alter testis function. Weused eight sheep, four Santa Ines and four crossbreed of Santa Ines/Dorper. After castration the animals gavethe Gonadosomatic index. Gauged to volume ratio of testis compartments using a reticle. The constituents of thetubular casing were evaluated: Lamina propria, seminiferous epithelium of the lumen and interstitialcompartment constituents: Leydig cells, blood vessels and connective tissue. Only the tubular diameter andheight of the seminiferous epithelium showed significant differences, being higher for sheep Santa Ines. It wasconcluded that there was little difference in testicular morphometry of animals studied and it was found that theracial cross does not induce significant changes in parameters between groups. (AU)

Histometria dos túbulos seminíferos em testículos de ovinos Santa Inês e mestiços Santa Inês e Dorper / Histometry of seminiferous tubules in testes of ovine Santa Ines and mongrel Santa Inês and Dorper

The morphometric measurements of the testicles is intended to measure the dimensions of constituenttestis parenchyma quantitatively evaluating spermatogenesis. This study aimed to analyze the spermatogenesisof sheep Santa Ines and crossbred Santa Ines and Dorper watching the racial crossing alter testis function. Weused eight sheep, four Santa Ines and four crossbreed of Santa Ines/Dorper. After castration the animals gavethe Gonadosomatic index. Gauged to volume ratio of testis compartments using a reticle. The constituents of thetubular casing were evaluated: Lamina propria, seminiferous epithelium of the lumen and interstitialcompartment constituents: Leydig cells, blood vessels and connective tissue. Only the tubular diameter andheight of the seminiferous epithelium showed significant differences, being higher for sheep Santa Ines. It wasconcluded that there was little difference in testicular morphometry of animals studied and it was found that theracial cross does not induce significant changes in parameters between groups.

MORFOLOGIA DA GÔNADA DE FILHOTES NATIMORTOS DE TARTARUGA-DE-PENTE (Erecmochelys imbricata)

As alterações climáticas são consideradas grande ameaça à biodiversidade global. As tartarugas de pente (Eretmochely imbricata) são as mais criticamente ameaçadas dentre as espécies que desovam o litoral do Rio Grande do Norte. Sabendo que as tartarugas marinhas exibem sua determinação sexual dependente da temperatura, são consideradas vulneráveis a mudanças nos regimes térmicos. Assim, o presente estudo tem como objetivo caracterizar a morfologia das gônadas de filhotes natimortos de tartaruga-depente. Os animais foram coletados na praia de Cabo de São Roque, Município de Barra de Maxaranguape. Após a coleta, retirou-se o plastrão, onde foi possível a identificação e dissecação do complexo gônada-mesonefro (CGM). Esse complexo foi submetido a análise macroscópica, posteriomente fragmentando e fixado com paraformaldeído a 4% e glutaraldeido a 2,5% em tampão fosfato por 24h para processamento para microscopia de luz e microscopia eletrônica de transmissão (TEM), respectivamente. Verificou-se que as gônadas das fêmeas possuíam um aspecto granuloso, cor esbranquiçada translúcida e forma fusiforme. Já nas gônadas masculinas observou-se uma superfície lisa, com cor branco opaco e formato aproximadamente ovoide. Também foi possível observar maior vascularização na superfície das gônadas das fêmeas em relação aos machos. Os testículos apresentaram a largura direita média de 63,48 ± 17,66m e esquerda de 6 1,11 ± 14,96m, enquanto os ovários direito 71,79 ± 17,67m e o esquerdo de 66,86 ± 20,74 m (P>0,05) Na microscopia, foi possível confirmar os sexos dos indivíduos, principalmente, pela diferença da estrutura da medula, onde as fêmeas possuíam celularidade desorganizada, com presença de ovogônias, lacunas e vasos sanguíneos. Os machos apresentaram um padrão organizacional na medula marcado pela presença de túbulos seminíferos em toda sua extensão. No que se refere a ultraestrutura, as fêmeas apresentaram um estroma marcado por uma rede de associação entre vasos sanguíneos, lacunas, fibroblastos, fibras de colágeno, músculo liso, células intersticiais com citoplasma rico em vesículas eletrodensas e células da linha germinativa (ovogônias) que apresentavam forma oval grande, com núcleo conspícuo e não central, citoplasma abundante. Nas masculinas foi possível observar no interior dos túbulos seminíferos, células piramidais com pouca heterocromatina, citoplasma apical aproximadamente triangular e basal com um nucléolo evidente característico de célula de Sertoli (CS). Junto a lâmina basal identificou-se células pavimentosas com características de células mioides (Cm) e no interstício observou-se células com características de função endócrina com vesículas eletrodensas em seu interior, padrão semelhante a células de Leydig (Ly). As Ly apresentavam núcleo arredondado, heterocromatina distribuída irregularmente e um nucléolo proeminente. Os ductos paramesonéfricos nas fêmeas é revestido por epitélio colunar simples e células cúbicas altas, Já nos machos, apresenta-se revestido por células cúbicas baixa, com perfil elíptico alongada e caracterizado pela ausência de lúmen. Os apêndices nas fêmeas apresentam um anel de ligação com oviduto com uma maior quantidade de células quando comparadas com o anel dos machos. Além disso, a extremidade dos apêndices também apresenta diferenças entre os sexos. Concluiu-se que tanto a avaliação macroscópica, microscópica e ultraestrutural se mostram técnicas efetivas e de confiança para a identificação sexual. Também vale salientar que na ausência das gônadas a análise microscópica dos ductos paramesonéfricos e dos apêndices dos ductos representam uma alternativa para a identificação sexual da espécie em questão.

Climate change is considered a major threat to global biodiversity. Hawksbill turtles (Eretmochely imbricata) are the most critically endangered species that spawn off the coast of Rio Grande do Norte. Knowing that sea turtles exhibit their temperaturedependent sex determination, they are considered vulnerable to changes in thermal regimes. Thus, the present study aims to characterize the gonad morphology of stillborn hawksbill turtle hatchlings. The animals were collected on the beach of Cabo de São Roque, municipality of Barra de Maxaranguape. After collection, the plastron was removed, where it was possible to identify and dissect the gonad-mesonephro complex (GMC). This complex was submitted to macroscopic analysis, later fragmented and fixed with 4% paraformaldehyde and 2.5% glutaraldehyde in phosphate buffer for 24h for processing for light microscopy and transmission electron microscopy (TEM), respectively. The female gonads were found to have a grainy appearance, translucent whitish color and spindle shape. In the male gonads, a smooth surface was observed, with an opaque white color and an approximately ovoid shape. It was also possible to observe greater vascularization on the surface of the female gonads compared to males. The testicles had an average right width of 63.48 ± 17.66m and the left 6 1.11 ± 14.96m, while the right ovaries 71.79 ± 17.67m and the left 66.86 ± 20.74 m (P>0.05) Under microscopy, it was possible to confirm the sex of individuals, mainly due to the difference in the structure of the marrow, where females had disorganized cellularity, with the presence of oogonias, lacunae and blood vessels. Males showed an organizational pattern in the medulla marked by the presence of seminiferous tubules throughout its length. Regarding the ultrastructure, the females presented a stroma marked by a network of association between blood vessels, gaps, fibroblasts, collagen fibers, smooth muscle, interstitial cells with cytoplasm rich in electron-dense vesicles and germline cells (ovogonia) that they had a large oval shape, with a conspicuous rather than a central nucleus, and abundant cytoplasm. In males, it was possible to observe inside the seminiferous tubules, pyramidal cells with little heterochromatin, approximately triangular and basal apical cytoplasm with an evident nucleolus characteristic of Sertoli cells (CS). Along the basal lamina, pavement cells with characteristics of myoid cells (Cm) were identified, and in the interstitium cells with characteristics of endocrine function were observed with electron-dense vesicles inside, a pattern similar to Leydig cells (Ly). Ly had a rounded nucleus, irregularly distributed heterochromatin and a prominent nucleolus. The paramesonephric ducts in females is lined with simple columnar epithelium and tall cubic cells, whereas in males, it is lined with low cubic cells, with an elongated elliptical profile and characterized by the absence of lumen. The appendages in females have an oviduct-connecting ring with a larger number of cells when compared to the ring in males. In addition, the end of the appendages also differs between sexes. It was concluded that both macroscopic, microscopic and ultrastructural evaluation are effective and reliable techniques for sexual identification. It is also worth noting that in the absence of the gonads, microscopic analysis of the paramesonephric ducts and duct appendages represent an alternative for the sexual identification of the species in question.

Avaliação da técnica de coloração AgNOR em testículos de ovinos / Evaluation of the AgNOR staining method in ovine testicles

A coloração pela prata das regiões organizadoras de nucléolos (NORs) é caracterizada por marcar proteínas ligadas ao ácido ribonucleico ribossômico, avaliando a proliferação em células normais ou neoplásicas. Objetivou-se estudar, em testículos de ovinos obtidos em matadouro, a validade do uso da técnica de coloração pela prata (AgNOR) na identificação das regiões organizadoras de nucléolo (NORs) em células saudáveis da linhagem espermatogênica. Utilizaram-se 43 pares de testículos de ovinos mestiços entre seis e 10 meses de idade. Testes de Wilcoxon e Spearman foram empregados, com nível de 5%. As médias das NORs nas células das gônadas direita e esquerda foram, respectivamente: espermatogônia (8,77±1,14 e 9,04±0,96), espermatócitos (4,99±2,00 e 6,20±2,07; P<0,05), Leydig (8,05±2,82 e 7,89±2,29) e Sertoli (8,07±1,88 e 7,61±2,16; P<0,05). Houve correlação (P<0,05) entre os lados para o número de NORs: espermatócitos x Leydig (0,49); espermatócitos x Sertoli (0,49) e Leydig x Sertoli (0,96). Conclui-se ser válido o emprego da técnica AgNOR para avaliar o potencial proliferativo das células saudáveis em testículos de ovinos com prática execução e baixo custo.(AU)

The silver staining technique for AgNOR nucleolar organizer regions (NORs) is characterized by marking proteins linked to the ribosomal ribonucleic acid, evaluating cell proliferation. The aim was to study the validity of the AgNOR staining technique in the testicular cell proliferation in crossbred ovine. Forty-three pairs of ovine testicles between 6 and 10 months old were collected. Wilcoxon and Spearman tests were used with a significance level of 5%. The mean NORs count in cells of the right and left gonads were respectively: spermatogonia (8.77±1.14 and 9.04±0.96), spermatocytes (4.99±2.00 and 6.20±2.07, P<0.05), Leydig (8.05±2.82 and 7.89±2.29) and Sertoli cells (8.07±1.88 and 7.61±2.16; P<0.05). There was a correlation between the mean values for the right and left sides for the number of NORs (P<0.05) between Leydig x spermatocytes (0.49); spermatocytes x Sertoli (0.49) and Sertoli x Leydig (0.96). The study demonstrates that the AgNOR staining technique is indicated to evaluate the cell proliferative potential in ovine testis with practical implementation and low cost.(AU)

Avaliação da técnica de coloração AgNOR em testículos de ovinos / Evaluation of the AgNOR staining method in ovine testicles

A coloração pela prata das regiões organizadoras de nucléolos (NORs) é caracterizada por marcar proteínas ligadas ao ácido ribonucleico ribossômico, avaliando a proliferação em células normais ou neoplásicas. Objetivou-se estudar, em testículos de ovinos obtidos em matadouro, a validade do uso da técnica de coloração pela prata (AgNOR) na identificação das regiões organizadoras de nucléolo (NORs) em células saudáveis da linhagem espermatogênica. Utilizaram-se 43 pares de testículos de ovinos mestiços entre seis e 10 meses de idade. Testes de Wilcoxon e Spearman foram empregados, com nível de 5%. As médias das NORs nas células das gônadas direita e esquerda foram, respectivamente: espermatogônia (8,77±1,14 e 9,04±0,96), espermatócitos (4,99±2,00 e 6,20±2,07; P<0,05), Leydig (8,05±2,82 e 7,89±2,29) e Sertoli (8,07±1,88 e 7,61±2,16; P<0,05). Houve correlação (P<0,05) entre os lados para o número de NORs: espermatócitos x Leydig (0,49); espermatócitos x Sertoli (0,49) e Leydig x Sertoli (0,96). Conclui-se ser válido o emprego da técnica AgNOR para avaliar o potencial proliferativo das células saudáveis em testículos de ovinos com prática execução e baixo custo.(AU)

The silver staining technique for AgNOR nucleolar organizer regions (NORs) is characterized by marking proteins linked to the ribosomal ribonucleic acid, evaluating cell proliferation. The aim was to study the validity of the AgNOR staining technique in the testicular cell proliferation in crossbred ovine. Forty-three pairs of ovine testicles between 6 and 10 months old were collected. Wilcoxon and Spearman tests were used with a significance level of 5%. The mean NORs count in cells of the right and left gonads were respectively: spermatogonia (8.77±1.14 and 9.04±0.96), spermatocytes (4.99±2.00 and 6.20±2.07, P<0.05), Leydig (8.05±2.82 and 7.89±2.29) and Sertoli cells (8.07±1.88 and 7.61±2.16; P<0.05). There was a correlation between the mean values for the right and left sides for the number of NORs (P<0.05) between Leydig x spermatocytes (0.49); spermatocytes x Sertoli (0.49) and Sertoli x Leydig (0.96). The study demonstrates that the AgNOR staining technique is indicated to evaluate the cell proliferative potential in ovine testis with practical implementation and low cost.(AU)

Efeitos de diferentes níveis de glicerina bruta na dieta de ovinos: avaliação morfofuncional testicular, renal e hepática.

Este estudo teve como objetivo avaliar os efeitos da ingestão de diferentes níveis de glicerina bruta contendo 80,5% de glicerol sobre a histomorfometria e histopatologia testicular, renal e hepática, níveis plasmáticos de testosterona e estresse oxidativo no parênquima testicular renal e hepático de ovinos. Foram utilizados 40 cordeiros mestiços da raça Santa Inês, machos, não castrados com 21 kg ± 0,8 kg de peso corporal inicial. Os animais foram submetidos casualmente aos níveis de 0% (Controle), 6% (G6), 12% (G12) e 18% (G18) de glicerina bruta com base na matéria seca. O peso, medidas biométricas do testículo, índice gonadossomático e peso líquido dos testículos foram maiores para os grupos tratados (P<0,05). Dentre os parâmetros tubulares, o comprimento total dos túbulos seminíferos foi maior no G6 (P<0,05). O volume tubular total e o volume do epitélio seminífero aumentaram para todos os grupos tratados (P<0,05). O volume individual da célula de Leydig aumentou no G12 enquanto a população da mesma célula por grama de testículo diminuiu (P<0,05), já no grupo G18 houve uma diminuição no diâmetro nuclear e volume individual da célula de Leydig, enquanto a população por grama de testículo aumentou (P<0,05), porém os níveis plasmáticos de testosterona não foram afetados. Segundo a análise histopatológica do parênquima testicular, os grupos experimentais sofreram descamação de epitélio seminífero e vacuolização de célula de Sertoli. Foram observados espermatócitos em necrose e presença de células gigantes em todos os grupos porém com graus diferentes. Nos grupos G12 e G18 foi observada intensa vacuolização das células de Sertoli. O grupo G12 teve o maior número de espermatócitos em paquíteno sofrendo necrose (P<0,05). O consumo de glicerina influenciou os marcadores do estresse oxidativo nos grupos G12 e G18, sendo o último o mais prejudicado. A produção de SOD aumentou nos grupos G12 e G18 (P<0,05). A produção de catalase aumentou nos grupos G12 e G18 (P<0,05). A produção de GST foi maior no grupo G18 (P<0,05). O nível de óxido nítrico diminuiu para todos os grupos tratados (P<0,05). A produção de MDA aumentou no G18 comparado aos grupos controle e ao G6 (P<0,05). A análise histomorfométrica do fígado revelou que os grupos G12 e G18 tiveram redução de núcleo e aumento de citoplasma de hepatócitos. Qualitativamente a esteatose foi a lesão predominante, os animais do grupo G18 foram os mais prejudicados. Os grupos G12 e G18 tiveram os maiores números de hepatócitos anucleados. Segundo a análise histomorfométrica renal o grupo G12 teve a maior dimensão de glomérulo e cápsula de Bowman, enquanto os grupos G6 e G12 tiveram os maiores valores de espaço capsular. Qualitativamente os grupos G12 e G18 tiveram maior grau de lesões, a glomerulonefrite proliferativa e membranosa foi predominante, porém a presença de glomérulos atrofiados também foi considerável. A análise de metabólitos de peroxidação lipídica no plasma sanguíneo revelou que os animais do G18 tiveram maiores níveis de MDA. Observou-se com os resultados expostos que a inclusão de até 6% de glicerina bruta na dieta de ovinos é segura, pois, não altera a histomorfometria nem histopatologia dos rins e fígado e melhora diversos aspectos que são indicativos de um bom desempenho reprodutivo do animal sem provocar qualquer estímulo ao estresse oxidativo, podendo assim, ser considerada como suplemento alimentar visando reduzir gastos com alimentação do rebanho sem qualquer prejuízo à saúde dos animais.

This study aimed to evaluate the effects of ingesting different levels of crude glycerin containing 80.5% glycerol on testicular, renal and hepatic histomorphometry and histopathology, plasma testosterone levels and oxidative stress in the renal and hepatic testicular parenchyma of sheep. Forty male Santa Inês crossbred lambs with 21 kg ± 0.8 kg of initial body weight were used. The animals were randomly subjected to levels of 0% (Control), 6% (G6), 12% (G12) and 18% (G18) of crude glycerin based on dry matter. Weight, biometric measurements of the testis, gonadosomatic index and net weight of the testicles were higher for the treated groups (P <0.05). Among the tubular parameters, the total length of seminiferous tubules was greater in G6 (P <0.05). The total tubular volume and the volume of the seminiferous epithelium increased for all treated groups (P <0.05). The individual volume of the Leydig cell increased in G12 while the population of the same cell per gram of testis decreased (P <0.05), whereas in the G18 group there was a decrease in the nuclear diameter and individual volume of the Leydig cell, while the population per gram of testicle increased (P <0.05), but plasma testosterone levels were not affected. According to histopathological analysis of the testicular parenchyma, the experimental groups suffered desquamation of seminiferous epithelium and vacuolization of the Sertoli cell. Spermatocytes were observed in necrosis and presence of giant cells in all groups but with different degrees. In groups G12 and G18, intense vacuolization of Sertoli cells was observed. The G12 group had the highest number of spermatocytes in pachytene undergoing necrosis (P <0.05). The consumption of glycerin influenced the markers of oxidative stress in groups G12 and G18, the latter being the most affected. SOD production increased in groups G12 and G18 (P <0.05). Catalase production increased in groups G12 and G18 (P <0.05). GST production was higher in the G18 group (P <0.05). The level of nitric oxide decreased for all treated groups (P <0.05). MDA production increased in G18 compared to control groups and G6 (P <0.05). Histomorphometric analysis of the liver revealed that groups G12 and G18 had a nucleus reduction and an increase in hepatocyte cytoplasm. Qualitatively steatosis was the predominant lesion, the animals in the G18 group were the most affected. Groups G12 and G18 had the highest numbers of anucleated hepatocytes. According to renal histomorphometric analysis, the G12 group had the largest dimension of the glomerulus and Bowman's capsule, while the groups G6 and G12 had the highest values of capsular space. Qualitatively, groups G12 and G18 had a higher degree of lesions, proliferative and membranous glomerulonephritis was predominant, but the presence of atrophied glomeruli was also considerable. The analysis of lipid peroxidation metabolites in blood plasma revealed that the animals in G18 had higher levels of MDA. It was observed with the exposed results that the inclusion of up to 6% of crude glycerin in the diet of sheep is safe, since it does not alter the histomorphometry or histopathology of the kidneys and liver and improves several aspects that are indicative of a good reproductive performance of the animal without provoking any stimulus to oxidative stress, and thus, it can be considered as a food supplement aiming to reduce spending on feeding the herd without any harm to the health of the animals.

Endocrine and paracrine regulation of zebrafish spermatogenesis: the Sertoli cell perspective

Spermatogonial stem cells (SSCs) either self-renew or differentiate into spermatogonia that further develop into spermatozoa. Self-renewal occurs when residing in a specific micro-environment (niche) While displacement from the niche would tip the signalling balance towards differentiation. Considering the cystic type of spermatogenesis in fish, the SSC candidates are single type A undifferentiated (Aund) spermatogonia, enveloped by mostly one niche-forming Sertoli cell. When going through a self-renewal cell cycle, the resulting new single type Aund spermatogonium would have to recruit another Sertoli cell to expand the niche space, while a differentiating germ cell cyle would result in a pair of spermatogonia that remain in contact with their cyst-forming Sertoli cells. In zebrafish, thyroid hormone stimulates the proliferation of Sertoli cells and of type Aund spermatogonia, involving Igf3, a new member of the Igf family. In cystic spermatogenesis, type A und spermatogonia usually do not leave the niche, so that supposedly the signalling in the niche changes when switching from self-renewal to differentiation. Recombinant zebrafish (rz) Fsh down-regulated Sertoli cell anti-müllerian hormone (amh) mRNA levels, and rzAmh inhibited differentiation of type A und spermatogonia as well as Fsh-stimulated steroidogenesis. Thus, for Fsh to efficiently stimulate testis functions, Amh bioactivity should be dampened. We also discovered that Fsh increased Sertoli cell Igf3 gene and protein expression; rzIgf3 stimulated spermatogonial proliferation and Fsh-stimulated spermatogenesis was significantly impaired by inhibiting Igf receptor signaling. We propose that in zebrafish, Fsh is the major regulator of testis functions and, supported by other endocrine systems (e.g. thyroid hormone), regulates Leydig cell steroidogenesis as well as Sertoli cell number and growth factor production to promote spermatogenesis.

Endocrine and paracrine regulation of zebrafish spermatogenesis: the Sertoli cell perspective

Spermatogonial stem cells (SSCs) either self-renew or differentiate into spermatogonia that further develop into spermatozoa. Self-renewal occurs when residing in a specific micro-environment (niche) While displacement from the niche would tip the signalling balance towards differentiation. Considering the cystic type of spermatogenesis in fish, the SSC candidates are single type A undifferentiated (Aund) spermatogonia, enveloped by mostly one niche-forming Sertoli cell. When going through a self-renewal cell cycle, the resulting new single type Aund spermatogonium would have to recruit another Sertoli cell to expand the niche space, while a differentiating germ cell cyle would result in a pair of spermatogonia that remain in contact with their cyst-forming Sertoli cells. In zebrafish, thyroid hormone stimulates the proliferation of Sertoli cells and of type Aund spermatogonia, involving Igf3, a new member of the Igf family. In cystic spermatogenesis, type A und spermatogonia usually do not leave the niche, so that supposedly the signalling in the niche changes when switching from self-renewal to differentiation. Recombinant zebrafish (rz) Fsh down-regulated Sertoli cell anti-müllerian hormone (amh) mRNA levels, and rzAmh inhibited differentiation of type A und spermatogonia as well as Fsh-stimulated steroidogenesis. Thus, for Fsh to efficiently stimulate testis functions, Amh bioactivity should be dampened. We also discovered that Fsh increased Sertoli cell Igf3 gene and protein expression; rzIgf3 stimulated spermatogonial proliferation and Fsh-stimulated spermatogenesis was significantly impaired by inhibiting Igf receptor signaling. We propose that in zebrafish, Fsh is the major regulator of testis functions and, supported by other endocrine systems (e.g. thyroid hormone), regulates Leydig cell steroidogenesis as well as Sertoli cell number and growth factor production to promote spermatogenesis.(AU)

Evaluation of quantitative parameters of Leydig cell in diabetic adults rats / Avaliação de parâmetros quantitativos de células de Leydig em ratos diabéticos adultos

The goal of present study was to evaluate the effects of diabetes on quantitative parameters of Leydig cells. Twelve adults Wistar rats were divided in: 1) Diabetic Group (DG), which was induced by a single intraperitoneal injection of streptozotocin (60 mg kg-1 of body weight); and 2) Control Group (CG), which received citrate buffer intraperitoneal. After eight weeks of diabetic induction, the animals were weighted, anesthetized and testicles were removed and routinely processed to paraffin embedded. Body weight (40%) and testicular weight (18%) of diabetic rats were significantly lower than control group.  Diabetic rats showed an increase in interstitial compartment but the tubular compartment did not differ. The individual volume of Leydig cells and nuclear diameter were lower in DG. However, the population of these cells was increased. In conclusion, diabetes induced by streptozotocin in adult rats promoted alterations in testicular compartments and changes on volume, nuclear diameter and population of Leydig cells, compromising the testicular function.(AU)

O objetivo do presente estudo foi avaliar os efeitos do diabetes nos parâmetros quantitativos de células de Leydig. Doze ratos machos adultos foram divididos em: 1) Grupo Diabético (GD) induzidos por injeção intraperitoneal única de estreptozotocina (60 mg kg-1 de peso corporal); e 2) Grupo Controle (GC) receberam tampão citrato, via intraperitoneal. Após oito semanas da indução, os animais foram pesados, anestesiados, os testículos foram removidos e processados rotineiramente em parafina. O peso corporal (40%) e testicular (18%) dos ratos diabéticos reduziu significativamente em relação ao grupo controle. Ratos diabéticos mostraram aumento no compartimento intersticial, mas o compartimento tubular não apresentou diferença significativa. O volume individual e o diâmetro nuclear de células de Leydig reduziram em GD. No entanto, a população dessas células aumentou. Em conclusão, o diabetes induzido por estreptozotocina, em ratos adultos, promoveu alterações nos compartimentos testiculares e mudanças no volume, diâmetro nuclear e população das células de Leydig, comprometendo a função testicular.(AU)

AVALIAÇÃO MORFOFUNCIONAL TESTICULAR DO LOBO-GUARÁ (Chrysocyon brachyurus) NAS DIFERENTES ESTAÇÕES CLIMÁTICAS DO ANO

Classificado como ameaçado de extinção pela IUCN e vulnerável pela Lista Nacional Espécies da Fauna Brasileira Ameaçadas de Extinção, o lobo-guará (Chrysocyon brachyurus) sofre com a redução e fragmentação do seu ambiente, atropelamentos, redução da população de suas presas naturais. Estudo sobre a diversidade genética dessa espécie relatou que mais de 40% das amostras obtida foram de animais mortos em estradas ou resgatados próximos de áreas urbanas com sinais de envenenamento e/ou machucados. Assim para o estabelecimento de técnicas em reprodução assistida e para a manutenção da variabilidade genética da espécie torna-se necessário o esclarecimento da dinâmica de funcionamento dos parâmetros testiculares ao longo do ano. Poucos estudos existem sobre a variação na produção de espermatozoide para carnívoros neotropicais, entretanto, foi identificaram uma variação nos parâmetros seminais ao longo do ano para o lobo-guará. Neste sentido, o estudo quantitativo dos componentes testiculares ao longo das estações climáticas torna-se essencial para o entendimento desse processo nessa espécie. Sendo assim, o objetivo deste trabalho foi avaliar a dinâmica do parênquima testicular do lobo-guará ao longo do ano. Foram avaliadas amostras testiculares de quatro indivíduos de lobos-guará para cada uma das diferentes estações climáticas do ano, estas amostras foram obtidas no acervo do Centro de Triagem de Animais Silvestres da Universidade Federal de Viçosa (CETAS-UFV), Viçosa, Minas Gerais. O volume testicular foi obtido através do cálculo do volume do elipsoide, e posteriormente foi realizada a confecção de lâminas histológicas. Os parâmetros morfométricos do testículo, como a proporção e o volume de túbulo seminífero, intertúbulo e de célula de Leydig, o comprimento tubular, o diâmetro de túbulo seminífero e de núcleo de célula de Leydig e o número de células de Leydig, foram mensurados ao longo das quatro estações climáticas do ano. O peso do testículo (6,03 ± 0,3g), a proporção (75,95 ± 5,84%) e o volume (4,58 ± 0,47ml) de túbulo seminífero mostraram-se maiores no outono, época de acasalamento, porém o volume de tecido intertubular não se alterou ao longo das diferentes estações climáticas. O diâmetro médio do túbulo seminífero na época acasalamento também foi maior (225,31 ± 6,07 µm) em relação ao período fora da época acasalamento. A proporção (40,00 ± 3,34%) e o volume (1,57 ± 0,38ml) de intertúbulo foram maiores no inverno, época de nascimento dos filhotes, o mesmo ocorre para o número de célula de Leydig totais, (9,44x108 ± 1,72x108), sendo o diâmetro e o volume do núcleo da célula o único componente que não teve variações ao longo do ano. Neste sentido é possível concluir que há uma sazonalidade nos constituintes do testículo de lobo-guará ao longo das diferentes estações climáticas do ano na região de estudo.

Classified as endangered by the IUCN and vulnerable by the National List Species of Brazilian Fauna Threatened of Extinction, the maned wolf (Chrysocyon brachyurus) suffers from the reduction and fragmentation of its environment, trampling, and population reduction of its natural prey. A study on the genetic diversity of this species reported that more than 40% of the samples obtained were from animals killed on roads or rescued near urban areas with signs of poisoning and / or bruising. Thus for the establishment of techniques in assisted reproduction and for the maintenance of the genetic variability of the species it is necessary to clarify the dynamics of testicular parameters functioning throughout the year. Few studies exist on the variation in sperm production for neotropical carnivores, however, a variation on the seminal parameters was identified for the maned wolf throughout the year. In this sense, the quantitative study of the testicular components throughout the climatic seasons becomes essential for the understanding of this process in this species. Thus, the objective of this work was to evaluate the dynamics of the testicular parenchyma of the maned wolf throughout the year. Testicular samples of four individuals of maned wolves were evaluated for each of the different climatic seasons of the year. These samples were obtained from the Centro de Triagem de Animais Silvestres da Universidade Federal de Viçosa (CETAS-UFV), Viçosa, Minas Gerais. The testicular volume was obtained by calculating the volume of the ellipsoid, and later the preparation of histological slides was performed. The morphometric parameters of the testis, such as the ratio and volume of seminiferous, intertubule and Leydig cell tubule, tubular length, seminiferous tubule diameter and Leydig cell nucleus, and Leydig cell number were measured at during the four seasons of the year. The weight of the testis (6.03 ± 0.3 g), the proportion (75.95 ± 5.84%) and the volume (4.58 ± 0.47 ml) of the seminiferous tubule were higher in the autumn, mating season, but the volume of intertubular tissue did not change over the different climatic seasons. The mean diameter of the seminiferous tubule at the mating season was also higher (225.31 ± 6.07 m) compared to the off-season mating period. The proportion (40.00 ± 3.34%) and volume (1.57 ± 0.38 ml) of intertubule were higher in the winter, the cub' birth time, the same for the total Leydig cell number, ( 9.44x108 ± 1.72x108), the diameter and volume of the cell nucleus being the only component that did not change throughout the year. In this sense, it is possible to conclude that there is a seasonality in the constituents of the testudo of maned wolf along the different climatic seasons of the year in the region of study.

Morfohistología testicular de Ceratophrys ornata (Bell) (Amphibia, Anura, Ceratophryidae) / Testicular morphohistology of Ceratophrys ornata (Bell) (Amphibia, Anura, Ceratophryidae)

En el presente trabajo se describen las características morfohistológicas del testículo de individuos adultos (n=5) de Ceratophrys ornata (Bell, 1843) provenientes de humedales del centro de Argentina. Los mismos se procesaron mediante técnicas histológicas de rutina, se cortaron a 8 µm y las láminas obtenidas se tiñeron con hematoxilina-eosina y tricrómico de Masson. Las gónadas son órganos pares, amarillentos, alargados y contorneados de 18,58 ± 0,23 mm de largo por 1,51 ± 0,13 mm de ancho. Histológicamente se observa una delgada túnica albugínea (6,29 ± 0,83 µm) rodeando a los testículos. En su interior se hallan lóculos seminíferos que miden 240,64 ± 38,52 µm de diámetro, en ellos se distinguen cistos con células espermatogénicas en distintas etapas de desarrollo. El tejido intersticial es escaso y en él se destacan las células de Leydig y vasos sanguíneos. Las espermatogonias I son las células más grandes de la serie germinal (20,03 ± 2,27 µm); poseen la cromatina granular y de aspecto multilobular, hallándose comúnmente una por cisto, estas originan a las espermatogonias II, más pequeñas (12,06 ± 1,14 µm). Los espermatocitos I presentan la cromatina levemente condensada y son un poco más chicos que sus precedentes (11,64 ± 0,36 µm). Los espermatocitos II miden 8,85 ± 0,54 µm. Las espermátidas I son esféricas, miden 5,95 ± 0,42 µm y se agrupan en cistos redondeados. Las espermátidas II, en cambio son alargadas y no se hallan dentro de cistos, pero siguen organizadas en paquetes asociadas a células de Sertoli. Los espermatozoides son células libres hacia el centro del lóculo, alargadas, flageladas y con una notable compactación nuclear. La morfohistología de los testículos analizados muestran características macroscópicas e histológicas similares a las observadas en otras especies de anfibios anuros neotropicales, presentando todas las células del linaje espermatogénico en un mismo lóculo, lo que indicaría que presentan ciclos espermatogénicos continuos.(AU)

This paper describes the morphohistological features of the testes of adult Ceratophrys ornata (Bell, 1843) (n = 5) from a wetland agroecosystem of central Argentina. The testes were processed by routine histological techniques, sectioned at 8 µm. The sheets obtained were stained with hematoxylin-eosin and Masson trichrome. The gonads are paired, yellowish, elongated contoured organs, 18.58 ± 0.23 mm long and 1.51 ± 0.13 mm wide. Histologically, there is a thin tunica albuginea (6.29 ± 0.83 µm) surrounding the testes. Inside them, there are seminiferous locules measuring 240.64 ± 38.52 µm in diameter, distinguished in cysts with spermatogenic cells at different stages of development. The interstitial tissue is scarce and the Leydig cells and blood vessels stand out. Spermatogonia I are the largest cells of the germinal series (20.03 ± 2.27 µm), have granular chromatin and multilobular aspect, commonly found in each cyst, and they originate spermatogonia II, which are smaller (12.06 ± 1.14 µm). Spermatocytes I have slightly condensed chromatin and are also slightly smaller than its predecessors (11.64 ± 0.36 µm). Spermatocytes II measure 8.85 ± 0.54 µm. Spermatids I are spherical, measuring 5.95 ± 0.42 µm and are grouped in rounded cysts. In contrast, the spermatids II are elongate and not within the cysts, but they are organized in packets associated with Sertoli cells. Sperms are free, elongated cells oriented towards the center of the loculus, flagellated, and with a remarkable nuclear compaction. The morphohistology of testes analyzed shows macroscopic and histologic features similar to those observed in other species of neotropical anuran amphibians, presenting all espermatogenic lineage cells in a single, indicating that they have continuous spermatogenic cycles.(AU)

Morfohistología testicular de Ceratophrys ornata (Bell) (Amphibia, Anura, Ceratophryidae) / Testicular morphohistology of Ceratophrys ornata (Bell) (Amphibia, Anura, Ceratophryidae)

En el presente trabajo se describen las características morfohistológicas del testículo de individuos adultos (n=5) de Ceratophrys ornata (Bell, 1843) provenientes de humedales del centro de Argentina. Los mismos se procesaron mediante técnicas histológicas de rutina, se cortaron a 8 µm y las láminas obtenidas se tiñeron con hematoxilina-eosina y tricrómico de Masson. Las gónadas son órganos pares, amarillentos, alargados y contorneados de 18,58 ± 0,23 mm de largo por 1,51 ± 0,13 mm de ancho. Histológicamente se observa una delgada túnica albugínea (6,29 ± 0,83 µm) rodeando a los testículos. En su interior se hallan lóculos seminíferos que miden 240,64 ± 38,52 µm de diámetro, en ellos se distinguen cistos con células espermatogénicas en distintas etapas de desarrollo. El tejido intersticial es escaso y en él se destacan las células de Leydig y vasos sanguíneos. Las espermatogonias I son las células más grandes de la serie germinal (20,03 ± 2,27 µm); poseen la cromatina granular y de aspecto multilobular, hallándose comúnmente una por cisto, estas originan a las espermatogonias II, más pequeñas (12,06 ± 1,14 µm). Los espermatocitos I presentan la cromatina levemente condensada y son un poco más chicos que sus precedentes (11,64 ± 0,36 µm). Los espermatocitos II miden 8,85 ± 0,54 µm. Las espermátidas I son esféricas, miden 5,95 ± 0,42 µm y se agrupan en cistos redondeados. Las espermátidas II, en cambio son alargadas y no se hallan dentro de cistos, pero siguen organizadas en paquetes asociadas a células de Sertoli. Los espermatozoides son células libres hacia el centro del lóculo, alargadas, flageladas y con una notable compactación nuclear. La morfohistología de los testículos analizados muestran características macroscópicas e histológicas similares a las observadas en otras especies de anfibios anuros neotropicales, presentando todas las células del linaje espermatogénico en un mismo lóculo, lo que indicaría que presentan ciclos espermatogénicos continuos.

This paper describes the morphohistological features of the testes of adult Ceratophrys ornata (Bell, 1843) (n = 5) from a wetland agroecosystem of central Argentina. The testes were processed by routine histological techniques, sectioned at 8 µm. The sheets obtained were stained with hematoxylin-eosin and Masson trichrome. The gonads are paired, yellowish, elongated contoured organs, 18.58 ± 0.23 mm long and 1.51 ± 0.13 mm wide. Histologically, there is a thin tunica albuginea (6.29 ± 0.83 µm) surrounding the testes. Inside them, there are seminiferous locules measuring 240.64 ± 38.52 µm in diameter, distinguished in cysts with spermatogenic cells at different stages of development. The interstitial tissue is scarce and the Leydig cells and blood vessels stand out. Spermatogonia I are the largest cells of the germinal series (20.03 ± 2.27 µm), have granular chromatin and multilobular aspect, commonly found in each cyst, and they originate spermatogonia II, which are smaller (12.06 ± 1.14 µm). Spermatocytes I have slightly condensed chromatin and are also slightly smaller than its predecessors (11.64 ± 0.36 µm). Spermatocytes II measure 8.85 ± 0.54 µm. Spermatids I are spherical, measuring 5.95 ± 0.42 µm and are grouped in rounded cysts. In contrast, the spermatids II are elongate and not within the cysts, but they are organized in packets associated with Sertoli cells. Sperms are free, elongated cells oriented towards the center of the loculus, flagellated, and with a remarkable nuclear compaction. The morphohistology of testes analyzed shows macroscopic and histologic features similar to those observed in other species of neotropical anuran amphibians, presenting all espermatogenic lineage cells in a single, indicating that they have continuous spermatogenic cycles.

Morphometry and histomorphometry of the testis in crossbred pigs fed diets with different protein levels / Morfometria e histomorfometria do testículo em suínos híbridos recebendo dietas com diferentes níveis de proteína

Aiming to evaluate the effect of the diet protein content on testicular parameters in pigs, 21 non-gelded male Dalland pigs were used and randomly divided into three groups. Males belonging to groups G2 and G3 received a diet with crude protein levels of 15% below and above, respectively, in relation to G1 (control). At 210 days of age, animals were castrated, and testis and epididymis were collected for morphometric and histomorphometry analyses. No difference was observed in relation to the total length of seminiferous tubules (G1=3239.9±333,3m; G2=2989.4±171,7m and G3=3059.5±254.9m), population of Sertoli cell (G1=4.7±0.5x10(9); G2=4.3±0.3x10(9) and G3=4.7±0.5x10(9)), population (G1=31.6±5.58x10(9); G2=27.3±4.0x10(9) and G3=26.4±3.9x10(9)) and volume of Leydig cells (G1=1289.3±182.6µm³; G2=1179.1±85.4µm³ and G3=1133.3±37.8µm³) and sperm production (G1=5.9±0.9x10(9); G2=5.6±0.6x10(9) and G3=5.1±0.3x10(9)). Protein levels were sufficient to maintain spermatogenesis in different experimental groups. It can be concluded that the magnitude of variation in levels of protein used in different stages of development was not sufficient to promote significant changes in testicular development and spermatogenesis process in adult animals.(AU)

Avaliou-se o efeito do teor de proteína da dieta sobre características testiculares em suínos, utilizando-se 21 suínos da raça Dalland, distribuídos aleatoriamente em três grupos. Os animais do G2 e G3 receberam dieta com porcentagens de proteína bruta de 15% para mais e para menos, respectivamente, em relação ao G1 (controle). Aos 210 dias de idade, os animais foram orquiectomizados e os testículos e epidídimos foram coletados para análises morfométricas e histomorfométricas. Observou-se efeito significativo da porcentagem de proteína sobre o comprimento e a largura dos testículos, e nenhuma diferença foi identificada em relação ao comprimento total dos túbulos seminíferos (G1=3239,9±333,3m; G2=2989,4±171,7m e G3=3059,5±254,9m), à população de células de Sertoli (G1=4,7±0,5x10(9); G2=4,3±0,3 x10(9) e G3=4,7±0,5x10(9)), à população (G1=31,6±5,58x10(9); G2=27,3±4,0x10(9) e G3=26,4±3,9x10(9)) e ao volume das células de Leydig (G1=1289,3±182,6µm³; G2=1179,1±85,4µm³ e G3=1133,3±37,8µm³) e à produção espermática (G1=5,9±0,9 x10(9); G2=5,6±0,6x10(9) e G3=5,1±0,3x10(9)). Os percentuais de proteína foram suficientes para a manutenção da espermatogênese nos diferentes grupos. Pode-se concluir que a magnitude da variação dos níveis de proteína usada em diferentes fases do desenvolvimento não foi suficiente para promover alterações significativas no desenvolvimento testicular e no processo espermatogênico em animais adultos.(AU)

Preparation of infertile male rabbits by local electron beam irradiation for intratesticular transplantation of autologous bone marrow stem cells

PURPOSE: To explore an efficient and safe protocol for the preparation of infertile male rabbits from which bone marrow stem cells (BMSCs) could be isolated and cultured. METHODS: Autologous BMSCs could be used for intratesticular transplantation and male infertility research. For this model, various doses (e.g., 6, 8, 10, or 12 Gy) of electron beam irradiation from a linear accelerator were locally applied to the scrotum of 5-month-old male New Zealand white rabbits. The effects of irradiation were compared between treatment groups, and with age-matched normal controls. Both morphology and hollow ratios of seminiferous tubules (HRST) were examined two, four, six, eight and 12-weeks post-irradiation. RESULTS: The seminiferous epithelium showed varying degrees of damage in all treatment groups compared with unirradiated controls, yet Sertoli and Leydig cells appeared unaffected. A dose-dependent response in spermatogenesis was also observed. BMSCs that were isolated and cultured from rabbits of the normal control group and the 12 Gy treatment group were compared with respect to morphology and growth. Starting at 6 weeks, HRST of the 12 Gy-treatment group were stable, and were the highest among all the groups. BMSCs from rabbits treated with 12 Gy also exhibited similar growth as the control group. CONCLUSION: Local dose of 12 Gy to the testes of 5-month-old male New Zealand rabbits is a protocol with which to obtain autologous bone marrow stem cells.(AU)