Stereological and morphological analysis of the effects of aging on spermatogenesis in rat testis

Abstract Purpose: To quantify, through stereological and morphometric analysis, spermatogenesis in rats undergoing the natural aging process. Methods: Seventy-two male Wistar rats were divided into 6 equal groups according to age at the time of killing: 3, 6, 9, 12, 18, and 24 months. All the rats were subjected orchiectomy and collection of testicular parenchymal fragments for histological and morphometric analysis. The numerical density of spermatids was calculated using a stereological study, and morphometric analysis was conducted to measure the height of the germinal epithelium and the area of the seminiferous tubules. Results: We found that the 18 and 24 months groups showed a significant reduction in the number of round spermatids. However, the height of the germinal epithelium was not significantly different between the groups. The area of seminiferous tubules was also significantly reduced in the elderly rats compared to that in the young ones. Conclusion: Aging of rats showed a significant reduction in the number of round spermatids and the area of the seminiferous tubules, more pronounced in the rats at 18 and 24 months of life.

Influence of prolonged flaxseed ( Linum usitatissimum ) consumption over epididymis and testicle histoarchitecture of Wistar rats / Influência do consumo prolongado de semente de linhaça ( Linum usitatissimum ) sobre a histoarquitetura dos testículo e epidídimo de ratos Wistar

Flaxseed is considered a functional food with several health benefits. However, because of its high phytoestrogen content, flaxseed influences hormone metabolism and affects the gonadal biomorphology. In this study, computerized histomorphometry was used to evaluate seminiferous and epididymal tubules, considering the different regions of the epididymis (head, body and tail) of rats subjected to a prolonged diet of flaxseed. Young adult male Wistar rats (n=20) were divided into 2 groups during their lactation period: Control Group (CG), fed casein-based meals and Flaxseed Group (FG), fed a 25% flaxseed meal. After 250 days of continuous ingestion, the animals were euthanized and a blood sample was collected. The testicles and epididymis were removed and fixed in buffered formalin solution. The samples were subjected to routine histological paraffin techniques and stained with hematoxilin and eosin. Immunostaining was performed using an antivimentin antibody for Sertoli cell identification. For morphometry, images of the slides were scanned and analyzed using Image J to determine the epithelial height, tubular and luminal diameter and tubular and luminal area. In the hormonal evaluation, FG had a higher serum concentration of estrogen (P=0.001), but no change was observed in the concentration of testosterone. The morphometric assay of seminiferous tubules and epididymal regions revealed no significant differences between the analyzed groups. Similarly, Sertoli cell quantification showed no significant differences in the FG (P=0.98). These results revealed that the continuous and prolonged intake of 25% flaxseed meals from gestation to 250 days of age, even with a significant increase in serum levels of estradiol, does not exert adverse effects on the testicular and epididymal structure or on the cells participating in the spermatogenesis of rats.(AU)

A semente de linhaça é considerada um alimento funcional com vários efeitos benéficos à saúde. Entretanto, devido ao seu elevado teor de fitoestrógenos, esta semente pode influenciar no metabolismo hormonal e interferir na biomorfologia gonadal. Neste estudo, utilizamos a histomorfometria computadorizada para avaliar os túbulos seminíferos e epididimários, considerando as diferentes regiões do epidídimo (cabeça, corpo e cauda) de ratos submetidos a uma dieta prolongada de semente de linhaça. Foram utilizados ratos Wistar machos adultos jovens (n=20) divididos em 2 grupos, durante o período de lactação: Grupo Controle (GC) a base de caseína e Grupo Linhaça (GL) alimentados com 25% de semente de linhaça. Ao final de 250 dias de ingestão contínua, os animais foram sacrificados e amostra de sangue foi coletada. Os testículos e epidídimos foram retirados e fixados em formol tamponado. As amostras foram submetidas ao processamento histológico de rotina para parafina e coradas em hematoxilina e eosina. Foi feita a imunomarcação com anticorpo antivimentina para identificação das células de Sertoli. Para morfometria, as imagens das lâminas foram digitalizadas e analisadas pelo software ImageJ para obtenção dos dados de altura epitelial, diâmetro e área tubular e luminal. Na avaliação hormonal o GL teve maior concentração de estrógeno sérico (p=0,001), mas nenhuma mudança na concentração de testosterona foi observada. Nos parâmetros morfométricos dos túbulos seminíferos e das regiões epididimárias, não houve diferenças significativas entre os grupos analisados. Da mesma forma, a quantificação das células de Sertoli não apresentaram diferenças significativas no GL (p=0,98). Estes resultados mostraram que o consumo contínuo e prolongado de 25% de semente de linhaça desde período gestacional até 250 dias de idade, mesmo com o aumento significativo nos níveis séricos de estradiol, não exerceram efeitos adversos sobre a estrutura testicular e epididimária, assim como nas células participantes da espermatogênese em ratos.(AU)

Aspects of spermatogenesis and microscopic testicular morphology in Greater Rhea, Rhea americana (Linnaeus, 1758) / Aspectos da espermatogênese e morfologia microscópica testicular em Emas, Rhea americana (Linnaeus, 1758)

The purpose of this study was to study the microscopic morphology of the testicular parenchyma of Rhea americana birds. Fifty-four 2.5±0.5 year-old male adults bred in captivity. were used. During commercial slaughter, samples of testis were collected in November/2005, December/2006 and May/2007, in order to compare possible differences. The samples underwent optical microscopy analysis and measurements of seminiferous tubule (ST) total diameters, lumen, epithelium thickness and the relative volume of parenchyma. The ST had circular form in transverse cross sections. November/2005 and December/2006 samples had many types of germinative cells and spermatozoa in lumen, but in May/2007 the samples of epithelium were poor regarding meiotic and mitotic pictures, and it was difficult to find any spermatozoon; in many tubules the lumen was inexistent or diminished. In December/2006 and May/2007 the averages were: tubule diameter 110.3 and 5.3mµ, lumen 52.4 and 4.5mµ, epithelium thickness 57.8 and 0.7mµ respectively. The volumetric proportions were: seminiferous epithelium 75.6 and 75.9, cysts in epithelium 2.1 and 1.0, ST 93.3 and 84.0, interstitium 6.2 and 15.6 respectively. The sperm reserves were: 19.7±2 and 0±0 x109 sperm cells in December 2006 and May 2007 respectively. Microscopic measures of seminiferous tubules, spermatic cells and diameter of the nuclei were presented. These data confirm reproductive seasonality, with breeding season in spring-summer with sperm production. A great variation n parenchyma, when compared breeding was noticeable.(AU)

O objetivo do estudo foi estudar a morfologia microscópica do parênquima testicular de emas (Rhea americana). Foram utilizados 54 machos adultos criados em cativeiro de 2,5±0,5 anos de idade. Durante o abate comercial foram coletadas amostras de testículos em novembro/2005, dezembro/2006 e maio/2007, para efeitos de comparação. As amostras foram processadas e para microscopia ótica de rotina para análise. Foram medidas diâmetro total de túbulos seminíferos (ST), lúmen, espessura do epitélio e a proporção volumétrica dos componentes do parênquima. O ST apresentou forma circular nas seções transversais. Em novembro/2005 e dezembro/2006, se observaram os tipos de células germinativas e espermatozoides no lúmen. Em maio/2007, as amostras de epitélio se observaram escassas meioses e imagens de mitose e era difícil de ver qualquer espermatozoide, em muitos dos túbulos o lúmen era inexistentes ou diminuído de tamanho. Em dezembro/2006 e maio/2007, as médias das características estudadas foram: diâmetro dos túbulos 110,3 e 5,3 mµ, lúmen 52,4 e 4,5mµ, espessura do epitélio 57,8 e 0,7mµ, respectivamente. As proporções volumétricas foram: epitélio seminífero 75,6 e 75,9, cistos no epitélio 2,1 e 1,0, túbulos seminíferos 93,3 e 84,0, interstício 6,2 e 15,6, respectivamente. Foram apresentadas medidas microscópicas de túbulos seminíferos, diâmetro dos núcleos das espermátides. Estes dados confirmam a sazonalidade reprodutiva, com época de reprodução na primavera - verão, com a produção de esperma. Foi perceptível uma grande variação nas medidas do parênquima testicular, quando se comparou a estação reprodutiva.(AU)

Morfologia do escroto, do testículo e das vias espermáticas de Metachirus nudicaudatus (Geoffroy, 1803), Didelphidae-Marsupialia / Morphology of scrotum and testicle, and spermatic pathways of Metachirus nudicaudatus (Geoffroy, 1803), Didelphidae-Marsupialia

Foram utilizadas as gônadas e vias espermáticas de cinco animais machos, adultos em fase reprodutiva, da espécie Metachirus nudicaudatus Geoffroy 1803, única espécie do gênero, para descrever a morfologia do escroto, do testículo e das vias espermáticas. O Metachirus possui escroto pré-peniano e que contém os testículos permanentemente. A pele escrotal é não pigmentada e com poucos pelos e glândulas. A lâmina parietal da túnica vaginal apresenta-se pouco pigmentada. Os testículos são ovais e ligados ao epidídimo através do pedículo testículo-epididimário. Eles são envolvidos, externamente, pela cápsula testicular e sustentados por um estroma de natureza conjuntiva. As células intersticiais são os elementos predominantes no abundante tecido intertubular. Os túbulos seminíferos são largos, enovelados e envolvidos por uma túnica própria fibroelástica, contendo células mióides. O epitélio seminífero é formado pelas células espermatogênicas e de Sertoli intercaladas. Os túbulos seminíferos convergem em direção à extremidade capitata do testículo, ficando revestidos por apenas células de sustentação, caracterizando uma região de transição entre túbulos seminíferos e túbulos retos, ocupada por uma estrutura tipo "válvula" que obstrui parcialmente o lume tubular. Os túbulos retos reúnem-se para formar um único dúctulo eferente, que percorre uma pequena extensão intratesticular, atravessa a albugínea e penetra no pedículo testículo-epididimário. A parte flexuosa do dúctulo eferente forma um lóbulo separado na parte medial do corpo do epidídimo. O epidídimo é envolvido pela cápsula epididimária e constituído pelo ducto epididimário, que se encontra bastante enovelado. O ducto epididimário é revestido por epitélio simples colunar pseudoestratificado apresentando células principais, basais, apicais e de "halo claro". As células principais são predominantes e apresentam características morfológicas e histoquímicas que diferem ao longo do ducto, possibilitando a caracterização de nove diferentes zonas epididimárias. É no lume da zona sete (início da cauda) que começa o pareamento de espermatozoides. Esse fenômeno coincide com alterações morfológicas bem evidentes e uma maior quantidade de mucossubstâncias neutras é secretada nessa zona.O ducto deferente apresenta-se dividido em três partes: justa-epididimária, funicular e abdominal, baseando nas variações histológicas e histoquímicas de seu epitélio e componentes envolventes. O ducto deferente não apresenta ampola e nem cruza o ureter antes de desembocar na uretra. O funículo espermático contém o ducto deferente, artéria e veias testiculares, vasos linfáticos, nervos e um desenvolvido músculo cremáster. Seus componentes apresentam modificações estruturais nas regiões proximal, média e distal, sendo notável a peculiar rede admirável.(AU)

Gonads and sperm pathways of five adult male Metachirus nudicaudatus in the reproductive phase were used to describe the morphology of scrotum, testicle, and spermatic tract. M. nudicaudatus has a scrotum pre-penis which contains the testicles permanently. The scrotal skin is not pigmented and has few hairs and glands. The parietal vaginal tunic is slightly pigmented. The testicles are oval and connected to the epididymis by testicular-epididymal pedicle; they are surrounded externally by the testicular capsule and supported by a stroma of connective nature. Interstitial cells are the predominant elements in abundant intertubular tissue. The seminiferous tubules are wide, meandering and surrounded by a fibro-elastic coat, containing myoid cells. The seminiferous epithelium is composed of spermatogenic cells and Sertoli cells interspersed. The seminiferous tubules converge toward the end of the testis capitata, getting coated only support cells, featuring a transition region between the seminiferous tubules and straight tubules, occupied by a type "valve" structure that partially blocks the tubular lumen. Straight tubules together to form a single efferent ductule, which runs a small intra-testicular extent, penetrates through the tunica and the pedicle testis-epididymis. The flexuosa part of the efferent ductule forms a separate lobe in the medial part of the body of the epididymis. The epididymis is enveloped by a capsule and epididymal comprising the epididymal duct, which is quite entangled. The epididymal duct is lined by pseudostratified columnar epithelium with simple principal, basal, apical and "clear halo" cells. The main cells are prevalent and have morphological and histochemical differing characteristics along the duct, enabling to characterize nine different epididymal areas. In the lumen of the seventh area (top of tail) that starts the pairing of sperm. This phenomenon coincides well with morphological change and a larger amount of neutral muco-substances is secreted in that area. Vas deferens has three parts: fair-epididymal, abdominal and funicular part, based on histological and histochemical changes of the epithelium and surrounding components. The vas deferens has no bulb and even crosses the ureter before flowing into the urethra. The spermatic cord contains the vas deferens, testicular artery and veins, lymphatic vessels, nerves and developed cremaster muscle. Its components have structural changes in the proximal, middle and distal region, with a peculiar admirable network.(AU)

Histological and morphometric analysis of testes of adult quail (Coturnix coturnix japonica) of Bangladesh / Análisis histológico y morfométrico de testiculos de codorniz adulta (Coturnix coturnix japonica) de Bangladesh

A histological and morphometric study was conducted to evaluate the seasonal testicular variations in the adult Japanese quail (Coturnix coturnix japonica) in Bangladesh. The weight of birds and testes, volume, length, circumference, seminiferous tubule diameter, circumference and height of germinal epithelium of testes were measured. Sections of testes obtained from studied birds were stained with H&E and images were taken by J software. Weight and volume of paired testes of quails significantly heavier (P<0.05) in long day period of summer and rainy season (March ­ September) than the short day period of autumn and winter (October ­ February). Seasons had no significant effect on live weight gain of quails. The weight and diameter of testes or height of the germinal epithelium of the seminiferous tubule were positively correlated (r=0.84; r=0.88). Spermatogenic activity was pronounced during summer and rainy season than that of autumn and winter (P<0.05).

Se realizó un estudio histológico y morfométrico para evaluar las variaciones testiculares estacionales de la codorniz japonesa adulta (Coturnix coturnix japonica) en Bangladesh. Se midieron el peso de las aves y sus testículos; el volumen, longitud, circunferencia y diámetro de los túbulos seminíferos, y la circunferencia y altura del epitelio germinal testicular. Secciones de testículos obtenidos de las aves estudiadas fueron teñidas con H&E y las imágenes fueron analizadas con el programa ImageJ. Se registró un aumento significativo en peso y volumen de los testículos de codornices (P<0,05) en el periodo día de verano y en la temporada de lluvia (marzo-septiembre) al período del día corto de otoño e invierno (octubre-febrero). El período estacionario no tuvo efecto significativo sobre el aumento de peso vivo de las codornices. El peso y el diámetro de los testículos, y la altura del epitelio germinal del túbulo seminífero se correlacionaron positivamente (r=0,84; r=0,88). La actividad espermatogénica fue más pronunciada durante el verano y durante la temporada de lluvias que en otoño e invierno (p<0,05).

Effects of human chorionic gonadotropin on the normal testicular tissue of rats

PURPOSE: To investigate the effects of human chorionic gonadotropin (hCG) on the testicular tissue of young male rats. METHODS: Male Wistar rats were assigned to groups (10 rats/group).Control Group received subcutaneous saline solution; Group 1 received hCG 50UI/Kg/dose; and Group 2 received hCG 100UI/Kg/dose, daily for 15 days. Half was submitted to bilateral orchiectomy on the 16th day and the other half 45 days after the beginning of the hormone application. Testicles were weighed, measured and has their volumes determined. The diameter of the tubules and the thickness of the seminiferous epithelium were measured. RESULTS: Control Group presented the highest values of testicles volume and weight. Rats in the Control presented normal histology. In G1 and G2 atrophy of the seminiferous tubules, apoptosis of germ cells and multinucleated giant cells were observed. Comparing groups, in the first operation Control rats had higher diameter values. In the second operation, the Control was only different from G1. As for thickness, Control had higher values in both operations. Comparing the time of operation, the diameter values were higher in G1 and G2 in the second operation. For all groups, the thickness of the epithelium was higher in the second operation. CONCLUSIONS: Human chorionic gonadotropin is gonadotoxic in rats. This effect was temporary and can affect reproductive potential. The total recovery of testicular damage in the studied range could not be proved, and the effects were not dose-dependent. .

Histological study of adult male rat seminiferous tubules following triclosan administation and the possible protective role of pomegranate juice

Triclosan [TCS] is an antimicrobial agent, widely incorporated in a variety of personal care products, household items, medical devices, and clinical settings. Recently, concern has been raised over TCS's potential for endocrine and reproductive disruption. The study aimed to elucidate the impact of TCS on the histological structure of the seminiferous tubules [STs] in adult male albino rats, as well as the possible protective role of pomegranate juice [PJ] coadministration. A total of 32 adult male albino rats [140-160 g] were randomly categorized into four equal groups. Group I [the control group]: rats in this group received PBS [1 ml/kg/day] orally. Group II: rats in this group received PJ orally at a dose of 10 ml/kg/day. Group III: rats in this group received TCS orally at a dose of 20 mg/kg/day. Group IV: rats in this group received TCS at the same dose as group III in conjunction with PJ daily. The experiment continued for 60 days. At the end of the experiment, blood samples were collected from the retro-orbital venous plexus of all rats for estimation of serum testosterone level. The animals were then euthanized. The testes of all rats were harvested for both light and transmission electron microscopic examination of the STs. The germinal epithelial height and the number of germ cells/high-power field [HPF] were estimated morphometrically in H and E-stained sections and statistically analyzed. The study revealed that PJ administration was safe as it did not alter serum testosterone levels as compared with the control group. Histologically, the STs of these animals exhibited normal appearance similar to that of the control group. TCS administration was associated with significantly lowered serum testosterone levels as compared with the control group. Histologically, the STs were lined with relatively few spermatogenic cells with deeply stained nuclei. Cytoplasmic vacuolation of the lining cells and exfoliation of germ cells in the tubular lumina were seen as well. Ultrastructurally, vacuolar degenerative changes involving all types of spermatogenic cells as well as Sertoli cells were revealed. Moreover, the germinal epithelial height and the number of germ cells/HPF were significantly reduced compared with the control group. Coadministration of PJ with TCS resulted in a significant increase in serum testosterone level as compared with the TCS group. Histologically, most of the STs retained normal appearance and epithelial stratification. Only some tubules revealed vacuolation of germ cells in the basal compartment with deeply stained nuclei. Mild ultrastructural alterations of germ cells were evidenced as well. These results were confirmed histomorphometrically by the significant increase in the germinal epithelial height and number of germ cells/HPF as compared with the TCS group. The study clearly indicates that TCS has the potential to adversely impact the testicular structure and function, and that PJ is able to ameliorate such adverse effects

Morphological, stereological and histometrical assessment of the testicular parameters between holstein and simmental bulls / Evaluación morfológica, estereológica e histométrica de los parámetros testiculares entre toros holstein y simmental

The testicular measurement is important criteria for experimental researches especially in toxicological studies and the prediction of spermatogenesis. In light of this knowledge, we aimed to estimate and compare these parameters in two different kinds of cattle breeds. The gross anatomical measurements were performed by vernier caliper, volume of the testis was estimated by Cavalieri method and seminiferous tubule diameter was measured on the histological sections by software-loaded computer attached to microscope. The mean testis weight, length, width, volume, and tubule diameter of the Simmental bulls and the Holstein bulls measured as 316 g, 12.1 cm, 6.9 cm, 295 cm3 and 226.68 um and 299 g, 12.1 cm, 6.8 cm, 285 cm3 and 223.44 µm, respectively. In conclusion all investigated parameters were not found statistically important in groups and between the breeds (p>0.05). The authors believed that the obtained data will contribute to the literature and facilitate future research.

La medición testicular es un criterio importante para las investigaciones experimentales sobre todo en los estudios toxicológicos y predicción de la espermatogénesis. El objetivo fue estimar y comparar estos parámetros en dos diferentes tipos de razas de ganado. Las mediciones anatómicas fueron realizadas con un pie de metro, el volumen de los testículos se estimó por el método de Cavalieri y el diámetro de los túbulos seminíferos se midió en las secciones histológicas observadas al microscopio mediante software. La Media del peso testicular, longitud, ancho, volumen y diámetro de los túbulos seminíferos de los toros Simmental fueron 316 g, 12,1 cm, 6,9 cm, 295 cm3 y 226,68 µm y de los toros Holstein fueron 299 g, 12,1 cm, 6,8 cm, 285 cm3 y 223,44 um, respectivamente. En conclusión, todos los parámetros investigados no tuvieron una importante significación en los grupos y entre las razas (p> 0,05). Creemos que los datos obtenidos contribuirán a la literatura y facilitar las futuras investigaciones.

Morphometric evaluation of seminiferous tubules in aged mice testes after melatonin administration

Melatonin, the pineal gland hormone as a direct or indirect antioxidant and free radical scavenger, is involved in the process of both aging and age-related diseases. This study investigates the effects of melatonin on the histology of testicular seminiferous tubules in aged mice. Twenty male, white mice, aged 16 months, that weighed 20-23 gr were equally divided into control and experimental groups. The experimental group was intraperitoneally injected with a daily single dose of 10 mg/kg melatonin for 14 days. The control group received only saline. Six days after the last injection, all mice were sacrificed and the testes were excised and processed for light microscope observation. In the morphometric study, we evaluated testicular seminiferous tubule parameters such as height of germinal epithelium, seminiferous tubule diameter, thickness of interstitial connective tissue and spermatogenesis index [SI]. SPSS software and student's t-test analyzed all parameters to assess the significance of changes between control and experimental groups. Melatonin-treated mice had seminiferous tubules with a wide lumen lined by low height germinal epithelium. The interstitial connective tissue thickened significantly in the experimental group [p<0.05], tubular diameter and germinal epithelium height decreased significantly [p<0.01], and the SI reduced compared to the control group [p<0.001]. The results of this study showed the disadvantages of melatonin on seminiferous tubules of aged mice testes

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.

The effects of Tynnanthus fasciculatus (Bignoniaceae) infusion on testicular parenchyma of adult Wistar rats

Traditional medicine provides strong guidance for scientific experiments involving plant products used by the Brazilian people. The species "cipó-cravo" (Tynnanthus fasciculatus) is a plant commonly used either to combat indigestion and stomachaches, or as a general stimulant and aphrodisiac. In this study, the effects of "cipó-cravo" infusion were investigated within the testicular parenchyma of adult Wistar rats. Rats were divided into 3 groups: a control (distilled water) and two treated groups, which received the plant infusion (100 and 200mg/animal/day). The 200mg dose promoted a significant increase of the testicular parenchyma weight and of the volume and total length of the seminiferous tubules, as well as in total daily sperm production and sperm production per gram of testis.

Aspectos morfológicos, morfométricos e histoquímicos dos túbulos armazenadores de espermatozoides da fêmea do peru Meleagris gallopavo / Histology, histochemistry, and morphometry of the tubuli spermatici of femeale turkeys, Meleagris gallopavo

Fragmentos da junção uterovaginal de 16 exemplares de fêmeas adultas de peru foram examinados, com o objetivo de determinar as modificações morfométricas e histológicas dos túbulos armazenadores de espermatozoides (SST), durante os períodos de atividade e repouso reprodutivo. Os estudos foram realizados a partir de dissecações do oviduto esquerdo, utilizando-se técnicas histológicas, histoquímica e histométricas de rotina. Essas estruturas originam-se de invaginações do epitélio das pregas da região uterovaginal e apresentam-se tubulosas, simples e não ramificadas, com epitélio formado por células prismáticas estreitas, claras e escuras, com núcleo vesiculoso basal e microvilos apicais. As secreções dessas células apresentaram reações negativas para PAS e Alcian Blue, indicando ausência de mucossubstâncias. Os parâmetros morfométricos utilizados mostraram que há diferenças significativas entre os períodos de atividade reprodutiva e de repouso. Na fase de atividade reprodutiva, o comprimento tubular, o volume tubular, o volume do lume, o número de SST na junção uterovaginal, o diâmetro externo e interno, a altura celular e o número de células por túbulos estão aumentados, e observam-se espermatozoides agrupados na luz do lume e células claras e escuras nos túbulos com vesículas supranucleares no citoplasma das células tubulares. Na fase de repouso, os parâmetros morfológicos estão reduzidos e não são encontrados espermatozoides na luz do lume. Nesta fase, não são evidenciadas as células escuras nem as vesículas supranucleares no citoplasma celular.

Fragments of the uterus-vaginal junction of 16 adult female turkeys were investigated aiming to determine the morphometrical and histological modifications in the tubuli spermatici (tubular sperm glands) during the breeding (August to March) and non-breeding (April to July) seasons. The studies were performed in dissected oviducts processed for histology, histochemistry, and morphometry of the tubuli spermatici (TS), structures which are originated from invaginations of the uterus-vaginal epithelium. TS are simple tubular glands, lined by a columnar epithelium formed by light and dark narrow cells, which presented euchromatic nucleus and apical microvillus. The TS epithelium secretion was negative for PAS and Alcian Blue stains, indicating the absence of mucosubstances. The tubular length, tubular volume, luminal volume, number of TS, as well as external and internal diameter, cellular height, and number of cells per tubule showed significant differences between the reproductive phases (breeding and non-breeding). In the breeding season, all these morphometrical parameters were increased when compared with the ones from the non-breeding season. Clusters of sperm in the lumen of the glands, as well vesicles in the cytoplasm of both light and dark epithelial cells, were observed in the breeding season. In the non-breeding season, luminal sperm, vesicles, and dark cells were not observed in the TS glands.

Seasonal variations in the histometric characteristics of the reproductive organs of pubertal West African dwarf bucks in their native tropical environment / Variaciones estacionales en las características morfométricas de los órganos reproductivos de la cabra enana púber macho del Oeste Africano en su medio ambiente nativo tropical

As ambient temperature and humidity are high all year round within clearly defined dry and rainy seasons in our humid tropical environment, we evaluated the effects of season on the histometric characteristics of the reproductive organs of pubertal West African dwarf bucks using 8 healthy animals. There were no significant differences (p>0.05) between the rainy and dry seasons in the basement membrane of the testes, seminiferous tubular diameter, and round spermatid nuclear diameter. There were also similarities (p>0.05) between the seasons in epididymal luminal diameters, epididymal epithelial heights as well as in Leydig/sertoli cell ratio and sertoli cell Índex. Quadratic regression analyses showed that all the histometric characteristics of the testes are highly predictable from the live weights of the animals. It is concluded from these results that the WAD goat would not have a restricted breeding season in it native tropical environment. Young sires could therefore be selected early in life for planned improvement programmes in research institutions and breeding centers thus providing farmers with quicker returns for their investments.

Como la temperatura ambiental y la humedad son altas durante todo el año dentro de estaciones secas y lluviosas claramente definidas en nuestro ambiente húmedo tropical, se evaluaron los efectos de las estaciones en la características histométricas de los órganos reproductivos de cabras enanas machos púber del Africa Occidental, usando 8 animales sanos. No hubo diferencias significativas (p> 0,05) entre las estaciones de lluvia y secas, en la membrana basal de los testículos, diámetro de los túbulos seminíferos, y el diámetro de la envoltura nuclear espermática. Sin embargo, hubo similitudes (p> 0,05) entre las diferentes estaciones en el diámetro luminal del epidídimo, altura del epitelio del epidídimo, así como en la proporción de células Leydig/Sertoli e índice de células de Sertoli. El análisis de regresión cuadrática mostró que todas las características histométricas de los testículos son altamente predecibles desde el peso de los animales vivos. Se concluye de estos resultados, que las cabras enanas del Africa Occidental no tendrían una restricción estacional para tener crías en el ambiente tropical nativo. Padres jóvenes por lo tanto, podrían ser seleccionados tempranamente en la vida para programas de mejora planeados en instituciones de investigación y centros criadores los que provean a granjeros de rápidos retornos por sus inversiones.

Stereological and morphometric analysis of collagen and seminiferous tubules in testes of patients with cryptorchidism submitted or not to treatment with human chorionic gonadotrophin

OBJECTIVES: Quantify the distribution of collagen and analyze the seminiferous tubules diameter in the testis of patients with cryptorchidism, to verify if the previous use of human chorionic gonadotrophin (hCG) affects these structures. MATERIALS AND METHODS: Samples of parenchymal tissue of cryptorchid testis obtained during peroperative biopsies were collected from 26 patients. Sixteen samples were embedded in paraffin and stained with picrosirius red to evidence fibers of collagen system. The quantification of these fibers was determined by stereological methods, using a test system M-42. To obtain seminiferous tubules diameter we used 10 of the 26 samples. These samples were embedded in Epon and the analyses were carried out in semi-thin sections, stained with toluidin blue. The selected results of each group were statistically analyzed and compared by the student's t and Tukey-Kramer's tests. RESULTS: The testicular interstitium and lamina propria of patients treated with hCG showed statistically significant less collagen system fibers, when compared to the testes of patients nontreated (0.30 percent versus 0.39 percent, p = 0.0079). The seminiferous tubules diameters were not statistically significant different between the testes of patients treated and nontreated with hCG (67.5 versus 59.35 µm, p = 0.0609). CONCLUSIONS: hCG use in the cryptorchidism could delay, at least temporarily, a progressive growth of fibers of collagen system. We did not find statistically significant difference in the seminiferous tubular diameters between treated and nontreated patients.

Quantificação de células dos túbulos seminíferos e rendimento da espermatogênese em cutias (Dasyprocta aguti) criadas em cativeiros / Histologic quantification of the seminiferous tubules cells and spermatogenesis yield in Agoutis (Dasyprocta aguti) rised in captivity

O presente estudo teve como objetivo avaliar o rendimento da espermatogênese de cutias criadas em cativeiro, por intermédio das razões encontradas entre tipos celulares do epitélio seminífero. Os resultados apontaram que o rendimento da espermatogênese da cutia dos nove aos quatorze meses de idade não chegou a um ponto de estabilização. O coeficiente de eficiência de mitoses espermatogoniais não aumentou com a idade. O rendimento meiótico, o rendimento geral da espermatogênese e o índice de células de Sertoli mostraram variações numéricas em função da idade, entretanto, não detectadas estatisticamente.

This study has as objective to evaluate the spermatogenesis yield of Agoutis rised in captivity, through the rates found between cellular types of the seminiferous epithelium. The results showed that the spermatogenesis yield of the Agoutis since 9 to 14 months of age did not reach the stabilization point. The coefficient of efficiency of the spermatogonium mitoses, did not increase with the age. The meiotic yield, usual spermatogenesis yield and the Sertoli cells index didn't showed numeric variation at function of the age, however, it was not detected by statistic data.

Acute effect of parathion on the seminiferous epithelium of immature mice

En el presente se estudió el efecto tóxico para la esparmatogenesis, de una inyección única del agropesticida organofosforado parathion, administrada a ratones inmaduros. Animales de 7 días de edad fueron inyectados por vía intraperitoneal con 1/3 de la LD50. Fue medido el peso corporal y testicular. Se efectúo un análisis cuali y cuantitativo de los túbulos seminíferos y de las poblaciones de células germinales en ratones controles y tratados, a los 8, 16, 28 y 50 días post-inyección. El efecto tóxico del pesticida se demuestra por una disminución del peso corporal y testicular y un daño temprano de las células germinales de los animales tratados. El efecto es reversible y la recuperación ocurre en los intervalos largos post-inyección

Histología testicular humana comparada, adulto joven y senil / Human testicular histology in young and senile men

La fisiología del envejecimiento implica cambios morfológicos que interfieren con la función testicular normal. El hombre senil mantiene la potencia fecundante, aunque su eficacia disminuye. Posiblemente, asociada a una presentación histopatológica testicular. Se utilizaron muestras de tejidos testicular de 3 individuos seniles (69 años), sometidos a orquiectomía terapéutica y de un joven adulto (25 años). Fallecido por causa desconocida. Las gónadas fueron procesadas por técnicas histológicas para Hematoxina-P.A.S. Al microscopio se evaluaron la morfometría, celularidad e histopatología del epitelio seminífero. En los resultados se obtuvo una reducción del diámetro tubular y altura del epitelio seminífero en los individuos seniles, al ser comparados con el individuo joven. Paralelamente, en los testículos de individuos seniles se describio una drástica disminución en el número de células de De Sertoli, y un poco menor para espermatogonias tipo A oscuras, A claras y B; con una relación gonia/Sertori alterada. Asociado a lo anterior, se encontró un porcentaje reducido de espermátidas redondas y alargadas y de espermatozoides, en lumen por túbulo. La evaluación histopatológica en los individuos seniles reveló un epitelio seminífero severamente dañado, con presencia de vacuolización, discontinuidad del epitelio y detención de la espermatogénesis. Por lo tanto, se concluye la existencia de un notorio efecto adverso del progreso de la edad hacia senil, en la estructura histológica testicular, involucrando las diferentes poblaciones celulares y la relación entre ellas, como también la integridad del epitelio seminífero

Stereological and ultrastructural study of seminiferous tubules in normal testes and in patients with varicocele

Testicular biopsies from fertile men and patients with varicocele were evaluated and examined by using the recent stereological technique and electron microscope. This study quantitated the seminiferous epithelium using the stereological methods. The numerical density of the germ cells was generally reduced in patients with varicocele when compared with that in the normal fertile men. However such reduction was not associated with a similar change in the supporting Sertoli cells. Moreover, the different ratios between various germ cells and Sertoli cells were statistically reduced in varicocele group. The seminiferous tubules were examined by light and electron microscope. The tissue architecture of the tubules in varicocele patients ranged from mild to severely altered. In affected tubules, spermatid nuclear and acrosomal morpology was abnormal. Sloughing of the individual germ cells as well as the vacuolization and degeneration of the Sertoli cell cytoplasm were evindent. Furthermore, Sertoli-spermatid junctional complexes appeared to be structurally abnormal, whereas Sertoli-Sertoli junctional cornplexes appeared to be structurally intact. On the basis of the results in this work, the Sertoli cell is, in fact, the primary intratubular site of alteration in varicocele patients leading secondarily to sloughing and spermatogenic disruption

Effect of H2-receptor antagonists, cimetidine and ranitidine on reproductive functions in male mice.

Na+,K(+)-ATPase and Ca(2+)-ATPase in testis were inhibited with an oral administration of cimetidine and ranitidine. Cimetidine at dose level of 100 and 30 mg while ranitidine at 70 and 10 mg per kg body wt inhibited the enzyme activities, 24 hr after single administration or daily administration for 15-days. Mg(2+)-ATPase activity was increased with cimetidine while ranitidine inhibited the enzyme. Michaelis-Menten kinetic characteristics revealed mixed type of inhibition for Na+,K(+)-ATPase with cimetidine, whereas it was noncompetitive for Ca(2+)-ATPase with cimetidine as well as ranitidine administration. Inhibition of Na+,K(+)-ATPase with ranitidine was also of noncompetitive type. Mg(2+)-ATPase behaved differently with administration of ranitidine at both the time points used i.e. noncompetitive type of inhibition after 24 hr and mixed type after 15-days. Histologically, signs of degeneration of testicular elements appeared after administration of cimetidine with a significant decrease in tubular diameter and germinal epithelial cell height. Ranitidine administration did not produce any change in the seminiferous tubules of testis. Scanning electron microscopy of spermatozoa from cimetidine-treated mice exhibited distinct departure from the normal morphology such as, (i) breaks at various places along distal portion of the tail, (ii) roughening, wrinkling and disorganization of plasma membrane of the head region, (iii) decapitation of the head and (iv) changes in shape of cytoplasmic droplet. Ranitidine administration showed normal morphology of the spermatozoa.

Observaçöes sobre a histoarquitetura geral dos testículos do hamster pós-natal / Observations on the general histoarchitecture of the post-natal hamster's testes

O presente trabalho é uma descriçäo sobre a morfologia da cápsula testicular e do complexo da rede testicular no hamster da variedade champanha, entre 10 e 90 dias de vida pós-natal. Adicionalmente, säo descritas algumas peculiaridades morfológicas dos túbulos seminíferos e do epitélio seminífero. Outras observaçöes apresentadas sobre a morfologia dos túbulos seminíferos e superficial da rede testicular