Expression of the homeobox proteins Dlx-5 and TLX1 during the development of cat testis / Expresión de las proteínas homeobox Dlx-5 y TLX1 durante el desarrollo del testículo de gato

SUMMARY: Recent studies have shown that homeobox proteins play an important role in the formation and development of tissues and organs in the embryonic period. In our study, the distribution of Dlx-5 and TLX proteins, which are members of the homeobox family, in the testis, epididymis and ductus deferens ducts of some cat breeds were investigated. For this purpose, in the study, 18 testes younger than six months (immature) and older than one year (mature) were examined under a light microscope using an immunohistochemical method (indirect streptavidin-biotin complex). While it was determined that Dlx-5 and TLX1 proteins were expressed at varying levels in cells in immature and mature cat testicles, epithelial cells of ductus epididymis and ductus deferens, and smooth muscle cells of ductus deferens, no differences were observed between cat breeds. Dlx-5 immunoreactivity was more intense in the testes, epididymis and deferens ducts of immature and mature compared to TLX1. These results suggested that both proteins play important roles in the development of male feline genital organs and in the secretion and differentiation of cells, and also further observation of Dlx-5 expression suggested that this protein may be more effective than TLX1 in testicular development and physiological processes.

RESUMEN: Estudios recientes han demostrado que las proteínas homeobox juegan un papel importante en la formación y desarrollo de tejidos y órganos en el período embrionario. En nuestro estudio, se investigó la distribución de las proteínas Dlx-5 y TLX, que son miembros de la familia homeobox, en los testículos, en el epidídimo y en los conductos deferentes de algunas razas de gatos. En el estudio fueron examinados, 18 testículos de animales menores de seis meses (inmaduros) y mayores de un año (maduros) bajo un microscopio óptico utilizando un método inmunohistoquímico (complejo indirecto de estreptavidina-biotina). Si bien se determinó que las proteínas Dlx-5 y TLX1 se expresaron en niveles variables en las células de los testículos de gatos inmaduros y maduros, las células epiteliales del epidídimo y del conducto deferente y las células del músculo liso del conducto deferente, no se observaron diferencias entre las razas de gatos. La inmunorreactividad de Dlx-5 fue más intensa en los testículos, epidídimo y conductos deferentes de gatos inmaduros y maduros en comparación con TLX1. Estos resultados sugieren que ambas proteínas tienen un rol importante en el desarrollo de los órganos genitales felinos masculinos y en la secreción y diferenciación de células, y también la observación de la expresión de Dlx-5 sugirió que esta proteína puede ser más efectiva que TLX1 en el desarrollo testicular y en los procesos fisiológicos.

Effects of vitamin e on the epithelial-mesenchymal transition in testicular development exposed to valproic acid / Efectos de la vitamina e en la interacción epitelio-mesenquimático en el desarrollo testicular expuesto a ácido valproico

SUMMARY: In testicular differentiation, somatic cells must adopt a specific destiny towards sustentacular, peritubular and interstitial cells, being fundamental for the morphogenesis of seminiferous tubules, mediated by morphogens such as Desert Hedgehog (DHH), insulin-like growth factor-1 (IGF-1) and fibroblastic growth factor 2 (FGF-2). Its alteration could be related to failures in the development mechanisms, such as those caused by valproic acid (VPA), which can be reversed with vitamin E (VE). The objective of the study was to evaluate the epithelial-mesenchymal transition (EMT) in the testicular development of mice exposed to VPA and VE. 12 groups of pregnant female mice were formed that were separated by days post-coital (dpc) at 12.5 dpc, 17.5 dpc and 6 weeks postnatal, each one subdivided into 4 groups of 5 pregnant women each. Subgroups received different treatments from the beginning to the end of gestation orally: 600 mg/kg of VPA, 600 mg/kg of VPA and 200 IU of VE, 200 IU of VE and the control group 0.3 mL of 0.9% physiological solution. Immunohistochemistry was performed for the detection of DHH, IGF-1 and FGF-2. Immunolocalization of DHH was observed in all stages, with more evident significant differences in integrated optical density (IOD) and percentage of immunoreaction area at 6 weeks postnatal, being lower in the VPA group. In IGF-1, lower intensity and distribution of immunostaining was observed in the fetal and pubertal stages in the VPA groups, a similar situation with FGF-2, but only evident at 17.5 dpc, with significant differences. These results demonstrate that VPA can alter EMT between somatic cells in testicular development, with VE being an agent capable of attenuating this process.

RESUMEN: En la diferenciación testicular, es necesario que las células somáticas adopten un destino específico hacia células sustentaculares, peritubulares e intersticiales, siendo fundamental para la morfogénesis de los túbulos seminíferos, mediado por morfógenos como Desert Hedgehog (DHH), Factor de Crecimiento Fibroblástico 2 (FGF-2) y Factor de Crecimiento símil a Insulina (IGF-1). Su alteración se podría relacionar a fallas en los mecanismos de desarrollo, como los que ocasiona el ácido valproico (VPA), los cuales pueden ser revertidos con la vitamina E (VE). El objetivo de estudio fue evaluar la transición epitelio-mesenquimática (EMT) en el desarrollo testicular de ratones expuestos a VPA y VE. Se conformaron 12 grupos de ratones hembra gestantes que se separaron por días post-coital (dpc) a los 12.5 dpc, 17.5 dpc y 6 semanas post-natal, cada uno subdividido en 4 grupos de 5 gestantes cada uno. Cada subgrupo recibió diferentes tratamientos desde el inicio hasta el término de la gestación vía oral: 600 mg/kg de VPA, 600 mg/kg de VPA y 200 UI de VE, 200 UI de VE y el grupo control 0,3 mL de solución fisiológica 0,9%. Se realizó técnica inmunohistoquímica para la detección de DHH, IGF-1 y FGF-2. Se observó la inmunolocalización de DHH en todos los estadios, con diferencias significativas más evidentes en la densidad óptica integrada (IOD) y porcentaje de área de inmunoreacción a las 6 semanas post-natal, siendo menor en el grupo VPA. En IGF-1, se observó en la etapa fetal y puberal menor intensidad y distribución de la marcación en los grupos VPA, situación similar con la inmunomarcación de FGF-2, pero sólo evidenciándose a los 17.5 dpc, con diferencias significativas. Estos resultados demuestran que el VPA puede alterar la EMT entre las células somáticas en el desarrollo testicular, siendo la VE un agente capaz de atenuar este proceso.

Concordancia en la medición testicular de varones adolescentes con 3 métodos de orquidometría / Concordance of testicular measurement in male adolescents with three methods of orchidometry

Introducción. El orquidómetro de Prader es el método estándar para medir el volumen testicular (VT) en niños y adolescentes. Objetivo. Evaluar la concordancia en la estimación del VT y del inicio puberal con las técnicas de orquidometría de Prader, Chipkevitch y Sotos. Métodos. Diseño descriptivo transversal realizado en varones de entre 9 y 20 años. Se midió el VT (ml) en cada adolescente con las técnicas de Prader (método de referencia), Chipkevitch (modelo gráfico) y Sotos (medición de ancho testicular con regla plástica y fórmula equivalente a ecuación elipsoide). Se excluyeron varones con patología urogenital y enfermedades que afectan el crecimiento testicular. Para la concordancia entre métodos, se utilizó kappa para el inicio puberal, y coeficiente de correlación intraclase (CCI) y gráficos de Bland-Altman (GBA) para el VT. Resultados. Se incluyeron 377 varones sanos. Para la concordancia en VT (ml), la comparación Prader-Chipkevitch obtuvo CCI: 0,994 y p < 0,001; y de CCI; 0,312 y p < 0,001 para la de Prader-Sotos. En los GBA se halló una media de las diferencias cercana a 0 ml en la comparación Prader-Chipkevitch y cercana a 8 ml en la de Prader-Sotos. El acuerdo en el inicio puberal obtuvo un valor de kappa 0,93 en la comparación Prader-Chipkevitch y de 0,75 en la de Prader-Sotos. Conclusión. Los orquidómetros de Prader y Chipkevitch tienen una excelente concordancia en la estimación del VT y el inicio puberal; por lo tanto, podrían intercambiarse en la atención diaria de varones adolescentes. El método de Sotos mostró una concordancia buena en la estimación del inicio puberal, pero baja en la medición del VT

Introduction. The Prader orchidometer is the standard method used to measure testicular volume (TV) in children and adolescents. Objective. To assess the concordance in the estimation of TV and puberty onset with the Prader, Chipkevitch, and Sotos orchidometric techniques. Methods. Cross-sectional descriptive study conducted among male children and adolescents aged 9-20 years. For each adolescent, TV was measured with the methods by Prader (gold standard), Chipkevitch (graphic model), and Sotos (measurement of testicular width with a plastic ruler and use of a formula equivalent to the ellipsoid equation). Male children and adolescents with urogenital conditions and disorders affecting testicular growth were excluded. Kappa statistics was used to determine concordance among methods for puberty onset, and intraclass correlation coefficient (ICC) and Bland-Altman (B&A) plots for TV. Results. In total, 377 healthy males were included. Regarding the concordance for TV (mL), the Prader-Chipkevitch comparison obtained an ICC of 0.994 and a p < 0.001; while the Prader-Soto comparison obtained an ICC of 0.312 and a p < 0.001. With the B&A plots, mean differences were close to 0 mL in the Prader-Chipkevitch comparison and close to 8 mL in the Prader-Sotos comparison. Concordance for puberty onset obtained a kappa value of 0.93 and 0.75 in the Prader-Chipkevitch and Prader-Sotos comparisons, respectively. Conclusion. The Prader and Chipkevitch orchidometers show an excellent concordance in estimating TV and puberty onset; therefore, both methods could be used interchangeably in the daily care of male adolescents. The Sotos method showed a high concordance in estimating pubertal onset, but low in measuring TV.

Gonadal maturation in Pseudoplatystoma metaense x Leiarius marmoratus hybrids, (Siluriformes: Pimelodidae) / Maduración gonadal en híbridos de Pseudoplatystoma metaense x Leiarius marmoratus (Siluriformes: Pimelodidae)

SUMMARY: Establishing guidelines for responsible management in fish production systems requires knowledge of the basic biology of the fish to be raised. The objective of this work was to determine the reproductive capacity of hybrids produced from the crossing of females of Pseudoplatystoma metaense with males of Leiarius marmoratus. Males presented a digitiform, unrestricted spermatogonial testicle containing caudal digits producing glycoproteins that do not form a seminal vesicle. It was possible to find free sperm in the lumen of the tubules and in the ducts. The ovary of the females was found to be saccular and synchronous with at least three groups of oocytes. In the first year of life, only oogonia up to previtellogenic oocytes (cortical alveolus 284.9 ± 35.7 mm in diameter) were found. After the second year vitellogenic oocytes 730 ± 3.78 mm in diameter were observed. The events of gonadal development of the hybrids indicate that they are gonochoric, synchronic animals. The maturation peaks in the high-water season, overlapping with the parental species. Therefore, the escape of hybrids from fish cultures to the rivers may increase the risk of crosses, gene introgression, or diminution of the reproductive capacity of the pure species.

RESUMEN: El establecimiento de pautas para la gestión responsable en los sistemas de producción de peces requiere el conocimiento de la biología básica de los peces a criar. El objetivo de este trabajo fue determinar la capacidad reproductiva de los híbridos producidos por el cruce de hembras de Pseudoplatystoma metaense con machos de Leiarius marmoratus. Los machos presentaron un testículo espermatogonial digital no restringido que contiene dígitos caudales que producen glucoproteínas que no forman una vesícula seminal. Fue posible encontrar esperma libre en la luz de los túbulos y en los conductos. Se encontró que el ovario de las hembras era sacular y sincrónico con al menos tres grupos de ovocitos. En el primer año de vida, solo se encontraron oogonia hasta ovocitos previtelogénicos (alvéolo cortical de 284,9 ± 35,7 mm de diámetro). Después del segundo año, se observaron ovocitos vitelogénicos de 730 ± 3,78 mm de diámetro. Los eventos de desarrollo gonadal de los híbridos indican que son animales sincrónicos gonocóricos. La maduración alcanza su punto máximo en la temporada de aguas altas, superponiéndose con las especies parentales. Por lo tanto, el escape de híbridos de cultivos de peces a los ríos puede aumentar el riesgo de cruces, introgresión genética o disminución de la capacidad reproductiva de las especies puras.

Histomorphometric analysis of gonads of green turtles (chelonia mydas) stranded on the coast of Espírito Santo state, Brazil / Análise histomorfométrica das gônadas de tartarugas verdes (Chelonia Mydas) provenientes de encalhe no litoral do Espírito Santo, Brasil

Studies on reproduction in sea turtles are important due to its life cycle, migratory patterns, high juvenile mortality and environmental impacts. This study aimed to analyse histomorphometrically gonads of C. mydas from the coastline of the Espírito Santo State, Brazil. Ovaries and testicles were collected between 2014 and 2015 from stranded animals. The material was fixed in formalin 10%, assessed macroscopically and processed for histomorphometrical evaluation. Gonads from 34 individuals were evaluated, twenty-four females and ten males. Macroscopic sexual identification presented 100% accuracy confirmed by histology. Sexual dimorphism was observed in one individual, which was considered as adult (CCL=1.023 m). Microscopy of female gonads revealed predominant previtellogenic follicles; oocyte diameter ranged between 161µm and 750µm and a positive correlation between ovarian length, largest oocyte and CCL was found. In males, autolysis was verified in five individuals. Viable testicles revealed predominant spermatogonia, primary spermatocytes and Sertoli cells in the seminiferous tubules and, Leydig cells and fibroblasts in the stroma. There was a positive correlation between tubular diameter and CCL and testicle length and CCL. Maturation of stromal tissue and a positive correlation between tubular lumen and CCL were also observed. Gonad development is proportional to individual growth.(AU)

Estudos em reprodução de tartarugas marinhas são importantes devido ao ciclo de vida, ao padrão migratório, à alta mortalidade juvenil e aos impactos ambientais. Objetivou-se analisar histomorfometricamente gônadas de C. mydas no litoral do Espírito Santo. Foram coletados ovários e testículos dessa espécie, entre 2014 e 2015. O material foi fixado em formol a 10% e avaliado macroscopicamente. Em seguida, foi processado para avaliação histomorfométrica. Foram avaliadas gônadas de 34 indivíduos, 24 fêmeas e 10 machos. Verificaram-se 100% de acurácia na identificação sexual à macroscopia, confirmada pela histologia. Observou-se dimorfismo sexual em um macho, que foi considerado adulto (CCC=1,023m). A microscopia dos ovários revelou folículos pré-vitelogênicos, cujos ovócitos apresentaram diâmetro médio entre 161µm e 750µm. Houve correlação positiva entre comprimento ovariano e diâmetro do maior ovócito e CCC. Nos machos, verificou-se autólise em cinco indivíduos. Os testículos viáveis revelaram espermatogônias, espermatócitos primários e células de Sertoli nos túbulos seminíferos, além de células de Leydig e fibroblastos no estroma. Houve correlação positiva entre diâmetro tubular e CCC e comprimento testicular e CCC. Verificou-se maturação do tecido estromal e correlação positiva entre o diâmetro do lúmen tubular e o CCC. Verifica-se que o desenvolvimento das gônadas é proporcional ao crescimento dos indivíduos.(AU)

Comb, cloaca and feet scores and testis morphometry in male broiler breeders at two different ages / Escores de crista, cloaca e de pé e morfometria testicular em galos de matriz pesada em duas diferentes idades

In broiler breeder roosters, the weight of the testes is positively associated with daily sperm production and fertility. In birds, the testes are located in the coelomic cavity, not being accessible to reproductive evaluation as mammalian testes. The reproductive evaluation of roosters is based on phenotypic traits. Any changes on testes will be reflected on fertility levels. The aim of this paper was to evaluate the potential of association of phenotypic traits as body weight (BW), comb scores (COS), cloaca scores (CLS) and feet scores (FS) with testicular morphometric parameters in adult Cobb® Mx roosters in two ages (25 and 45 weeks of age). The low BW roosters had lower testes weight (TW), smaller seminiferous tubule diameter (STD) and lower comb and cloaca scores than the medium and high BW roosters (P≤0.05). Heavy roosters had the highest COS and bigger seminiferous epithelium height (SEH). There was no statistical difference between the BW categories in FS and volumetric proportion of seminiferous tubule (ST) and interstitial tissue (IT). Considering the age effect, roosters with 45 weeks of age had lower TW and ST than 25-week roosters. Positive, moderate and significant correlation was found between testes weight and phenotypic characteristics as BW, COS and CLS in 45week roosters. A positive, moderate and significant correlation was found between body weight and histologic testicular characteristics as STD and SHE in both evaluated ages. In conclusion, there is a regression in the testes weight with age and it is more intense in Cobb® Mx roosters with lower BW. Furthermore, it is concluded that the BW positively influences the testes weight and histological quality of the testes. COS and CLS can be used with moderate potential prediction to identify roosters with low testes weight at 45 weeks of age.(AU)

Em galos de matriz pesada sabe-se que o peso dos testículos pode ser associado positivamente com a produção diária de espermatozoides e que alterações nos testículos irão refletir na fertilidade. Nas aves, os testículos estão localizados dentro da cavidade celomática, e, portanto, não são acessíveis para exame andrológico direto, como em mamíferos. A avaliação reprodutiva de galos se baseia principalmente em características fenotípicas. Buscou-se avaliar o potencial de associação de alguns parâmetros fenotípicos, como peso corporal (PC), escores de crista (ECR), escores de cloaca (ECL) e escores de pés (EP), com parâmetros morfológicos de testículos em galos Cobb® Mx em duas idades (25 e 45 semanas de idade). Galos leves apresentaram menor peso testicular (PT), menor diâmetro de túbulo seminífero (DT), e menores ECR e ECL que galos médios e pesados (P≤0.05). Galos pesados apresentaram os maiores ECR e maiores valores para altura do epitélio seminífero (AE), e DT. Não houve diferença estatisticamente significativa entre as categorias de peso para escores de pé e proporções de túbulo seminífero (TS) e interstício (INT). Considerando-se o efeito da idade, galos com 45 semanas de idade apresentaram menores PT e TS que galos com 25 semanas de idade. Correlação positiva, moderada e significativa foi encontrada entre peso de testículo e características fenotípicas como ECR, ECL e PC em galos de 45 semanas de idade. Foi encontrada correlação positiva, moderada e significativa entre PC e características testiculares como DT e AE nas duas idades avaliadas. Conclui-se que em galos Cobb® Mx ocorre regressão do testículo com o avanço da idade, e ela é mais intensa em galos com menor peso corporal. Conclui-se que o PC tem influência positiva no peso e índices histomorfométricos dos testículos, e que escores de crista e de cloaca podem ser utilizados com potencial moderado de predição para identificar galos com baixo peso de testículos em galos com 45 semanas de idade.(AU)

Interacciones epitelio-mesenquimáticas en el desarrollo testicular / Epithelial-mesenchymal transitions in the development of testis

RESUMEN: La espermatogénesis es un proceso continuo que se inicia durante el desarrollo embriofetal. Las relaciones auto, para y yuxtacrinas indican la interdependencia de las células intersticiales (de Leydig) con las células peritubulares (lamina propia) y células sustentaculares (de Sertoli). Ciertos morfógenos son fundamentales en este proceso. Las células sustentaculares son capaces de regular la diferenciación y función de las células peritubulares e intersticiales a través de la producción de IGF1, TGFA, TGFB y DHH. Las células peritubulares son capaces de producir P-Mod-S, regulando la diferenciación de las células sustentaculares, y a través de FGF2 y FGF9 modulan las transiciones epitelio-mesenquimática entre células sustentaculares y mesonefros. También remodelan la membrana basal del condón testicular y regulan la diferenciación y función de las células intersticiales por medio de IGF1, TGFA y TGFB. Las células intersticiales son las reponsables de la producción de testosterona e INSL3, influyendo en la diferenciación sexual masculina. Se plantea que provienen de células mesenquimales del epitelio celómico y mesonefros. Sin embargo, otros autores proponen su origen a partir de células de la cresta neural. Estas influyen a través de mecanismos paracrinos en la proliferación de las células sutentaculares por medio de activina A, teniendo como resultado la expansión del cordón testicular. Las interacciones entre las distintas poblaciones celulares a través de morfógenos inducen una transición epitelio-mesénquima fundamental en la formación y diferenciación de la gónada masculina.

SUMMARY: Spermatogenesis is a continuous process which starts during the embryo-fetal development. Auto, para and juxtacrine relations indicate the interdependence of the interstitial cells (Leydig) with the peritubular cells (lamina propria) and sustentacular cells (Sertoli). Certain morphogens are fundamental in this process. Sustentacular cells are able to regulate differentiation and function and peritubular interstitial cells through production of IGF1, TGFA, TGFB and DHH. Peritubular cells are able to produce P-Mod-S regulating differentiation sustentacular cells and through FGF2 and FGF9 modulate epithelial-mesenchymal transitions between sustentacular cells and mesonephros. They also remodel the basal membrane of the testicular condom and regulate the differentiation and function of the interstitial cells by means of IGF1, TGFA and TGFB. Interstitial cells are responsible for the production of testosterone and INSL3, influencing male sexual differentiation. It is suggested that they come from mesenchymal cells of the coelomic epithelium and mesonephros. However, other authors propose their origin from cells of the neural crest. These influence through paracrine mechanisms proliferation sutentaculares cells by activin A, resulting in the expansion of cord testicular. The interactions between the different cell populations through morphogens induce a fundamental epithelial-mesenchymal transition in the formation and differentiation of the male gonad.

Gonadal development of Peprilus medius (Peters, 1869) (Perciformes: Stromateidae) from Southeast of the Gulf of California, Mexico / Desarrollo gonádico de Peprilus medius (Peters, 1869) (Perciformes: Stromateidae) del Sureste del Golfo de California, México

The gonadal development assessment is a fundamental work in researching the reproduction of Peprilus medius, on the way to understand its population dynamics and to achieve a sustainable management of its fisheries. The diagnosis based on morphochromatic features is the quicker, more practical and cheaper method, and properly validated, it may be an accurate procedure. This is particularly useful in researches when it is required to provide prompt and accurate information to support decision-making for fisheries management, and during long-term fisheries monitoring to detect changes in reproductive patterns due to environmental anomalies. These are issues of concern in P. medius and other fishery resources. In this context, a scale that describes the gonadal development through morphochromatic and histological features is presented, and plus anatomical observations of the reproductive organs. Ovarian development was characterized through six phases: immature, development (sub-phases: early and late), mature, spawning (sub-phases: partial and partial post-spawning), post-spawning and resting. Testicular development was described through five phases: immature, development, ejaculation, ejaculated and resting. The position and shape of the reproductive organs are apparently defined by the shape of the fish and the visceral cavity. P. medius's gonads show a gonadal lobe at lower position and an upper lobe in the visceral cavity, and both lobes presented a horizontal placement from gonopore toward the back of the fish. In addition, gonadal deformities occurrence due to deformities in the visceral cavity was evidenced. We recommend to use presented descriptions as a guide in gonadal assessment.

La evaluación del desarrollo gonádico es una labor fundamental para investigar la reproducción de Peprilus medius, en vías a comprender la dinámica de sus poblaciones y lograr un manejo sustentable de sus pesquerías. El diagnóstico a partir de características morfocromáticas es el método más rápido, práctico y barato, y adecuadamente validado puede ser un procedimiento preciso. Este es particularmente útil en investigaciones cuando se requiere aportar información rápida y precisa que apoye la toma de decisiones de manejo pesquero, y durante los monitoreos pesqueros de largo plazo, que permiten detectar cambios en los patrones reproductivos debido a anomalías ambientales. Estos son temas preocupantes en P. medius y otros recursos pesqueros. En ese contexto se presenta una escala que describe el desarrollo gonádico a través de características morfocromáticas e histológicas, y además se presentan observaciones anatómicas de los órganos reproductores. El desarrollo ovárico fue caracterizado a través de seis fases: inmadura, desarrollo (subfases: inicial y avanzado), maduro, desove (subfases: parcial y postdesove parcial), postdesove y reposo. El desarrollo testicular fue descrito a través de cinco fases: inmadura, desarrollo, eyaculación y reposo. La posición y forma de los órganos reproductores están aparentemente definidos por la forma del pez y su cavidad visceral. Las gónadas de P. medius muestran un lóbulo gonádico en posición inferior y uno superior en la cavidad visceral, y ambos lóbulos presentan una colocación horizontal desde el gonoporo hacia la parte posterior del pez. Además se evidencia la ocurrencia de deformidades en las gónadas debido a deformidades en la cavidad visceral. Recomendamos utilizar las descripciones presentadas como una guía en la evaluación de las gónadas.

Musk gland seasonal development and musk secretion are regulated by the testis in muskrat (Ondatra zibethicus)

BACKGROUND: The muskrat is a seasonal breeder. Males secrete musk to attract females during the breeding season. The testosterone binding to the androgen receptor (AR) in musk glands of muskrat may play an important role conducting the musk secretion process. METHODS: The musk gland, testis and blood samples of musk rats are collected in both breeding and non-breeding seasons. Some part of the samples are kept in liquid nitrogen for transcriptome analysis and Western blotting test. Some part of the samples are kept in 70% alcohol for histology experiment, blood samples are kept at -20 °C for the serum testosterone measurement experiment. RESULTS: This study demonstrates that the quantity of secreted musk, the volume of the musk glands, the diameter of the gland cells and AR expression are all higher during the breeding season than at other times (p < 0.01). StAR, P450scc and 3ß-HSD expression in the Leydig cells of the testis were also higher during this season, as was serum testosterone. AR was also observed in the gland cells of two other musk-secreting animals, the musk deer and small Indian civet, in their musk glands. These results suggest that the testes and musk glands co-develop seasonally. CONCLUSION: The musk glands' seasonal development and musk secretion are regulated by the testes, and testosterone plays an important role in the seasonal development of musk glands.

Effect of high altitude hypoxia on the postnatal development of rat testis: histological and histometric study / Efecto de la hipoxia a gran altitud en el desarrollo postnatal en testículos de rata: estudio histológico e histométrico

The study was carried out at two different altitudes in the southern region of Saudi Arabia: Abha, 2,800 meters above sea level, the high altitude (HA) area and Jazan, 40 meters above sea level the low altitude (LA) area. Following exposure to high altitude, and up to the third week of postnatal development, some of the seminiferous tubules showed detachment from the basal lamina, spaces of different sizes within the epithelial layer with vacuoles in the center and Pyknotic nuclei were noted in the spermatogonia and primordial germ cells. Rounded spermatids were seen in the lumen only on day 35 of hypoxic group, no spermatozoa were recognized until day 45 of postnatal development. On day 45 hypoxic rat testes revealed various types of atrophy and degeneration in the seminiferous tubules and in the interstitial tissue, there was detachment of the basal laminae of the tubules and a profound decrease in cellularity. Significant decrease in epithelial height was noticed in these animals (P <0.05). Also, the diameter of the tubules showed slight decrease with concomitant increase in interstitial spaces in all hypoxic rats.

El estudio se llevó a cabo en dos sitios de altitud diferentes en la región sur de Arabia Saudita: en Abha, a 2.800 metros sobre el nivel del mar, la zona de gran altitud (GA) y en Jazan, a 40 metros sobre el nivel del mar, la zona de baja altitud. Después de la exposición a GA, y hasta la tercera semana del desarrollo postnatal, se observaron espermatogonias y células germinales primordiales en algunos túbulos seminíferos, un desprendimiento de la lámina basal, espacios de diferentes tamaños dentro de la capa epitelial, con vacuolas en el centro y núcleos picnóticos. Se encontraron espermátidas redondeadas a nivel del lumen en el día 35 de la hipoxia, y no se observaron espermatozoides hasta el día 45 del desarrollo postnatal. En el día 45, los testículos de las ratas hipóxicas revelaron varios tipos de atrofia y degeneración en los túbulos seminíferos y el tejido intersticial; no hubo separación de las láminas basales de los túbulos y se registró una profunda disminución de la celularidad. Además, se observó una disminución significativa en la altura del epitelio de estos animales (P <0,05). El diámetro de los túbulos mostró una ligera disminución con el aumento concomitante en los espacios intersticiales en todas las ratas hipóxicas.

Postnatal developmental anatomy of testes and epididymis of Gaddi goats / Desarrollo anatómico postnatal de los testículos y epidídimo de cabras Gaddi

The study revealed that the testes of one-day-old Gaddi goats have descended in the scrotum along with adnexa. They were elongated compressed latero-medially and became ovoid during their postnatal growth and morphogenesis. The two testes were not different significantly in their weight and measurement. The average weight which was about 2.9 gm at birth grew to 107 gm (40X) in pubertal animals and nearly maintained it during the post-puberty (113 g). The mean length (1.46 cm), width (0.85 cm) and thickness (0.62 cm) also grew by 47, 60, and 83 times, respectively. In post-pubertal animals it measured 7.06 x 5.20 x 5.10 cm. The study indicated a very fast growth of the testes from birth to puberty, which did not alter significantly after that. A relatively much faster growth in thickness and width over the length was the reason behind the change of the morphology from more elongated shape at birth to ovoid shape on maturity. The mediastinum testis was grossly discernible in the testis even at birth. Epididymis was identifiable into caput, corpus and cauda. The gross linear mensuration of these components did not differ in the caput and cauda regions on either side, whereas the length of corpus was more on the left side in all age group of animals. The growth curve revealed a continuous growth of all segments, but the fastest growth occurred in the early postnatal life (at 12­18 month age).

El estudio reveló que los testículos de las cabras Gaddi, de un día de edad, descienden al escroto junto con sus anexos. Se elongaron en dirección latero-medial y convirtieron en ovoides durante su crecimiento postnatal y en la morfogénesis. Los dos testículos no fueron significativamente diferentes en su peso y medidas. El peso promedio fue aproximadamente de 2,9 g al nacer, y aumentó a 107 g (40X) en animales púberes y casi mantuvo su peso durante la post-pubertad (113 g). La longitud (1,46 cm), ancho (0,85 cm) y espesor (0,62 cm) también crecieron unas 47, 60 y 83 veces, respectivamente. En los animales post-puberales las medidas fueron de 7,06 x 5,20 x 5.10 cm. Se observó un crecimiento muy rápido de los testículos desde el nacimiento hasta la pubertad, sin alterarse de manera significativa posteriormente. Un crecimiento relativamente mucho más rápido en espesor y anchura, sobre la longitud, fue la razón detrás del cambio de la morfología desde la forma más alargada, presente en el nacimiento, a la forma ovoide de la madurez. El mediastino testicular fue claramente identificado en el testículo, inclusive en el nacimiento. En el epidídimo se identificaron las porciones de la cabeza, cuerpo y cola. La medición lineal de estos componentes no difirió en las regiones de la cabeza y cola de cada lado, mientras que la longitud del cuerpo fue mayor en el lado izquierdo en todos los grupos. La curva de crecimiento reveló un crecimiento continuo de todos los segmentos, pero el mayor crecimiento se produjo en la vida postnatal temprana (entre los 12 y 18 meses).

Testicular and epididymal ltrasonography in Santa Inês lambs raised in Brazil

The objective of this work was to study, through ultrasonographic evaluation, changes in testes and epididymides of clinically healthy, peripubertal and pubertal Santa Inês lambs raised in Brazil. Periodic e valuations of weight, biometric characteristics (scrotal circumference, width and length) and ultrasound examinatio ns of the testes and epididymides of 20 lambs were performed between 84 and 280 days old at intervals of 28 days. Scans were performed in the sagittal, transverse, frontal and oblique planes to evaluate the echotexture of the testicular parenchyma and mediastinum and the tail epididymis as well as the thickness and width of the mediastinum testis. The testicular parenchyma demonstrated a homogeneous echogenicity patter n ranging from low to moderate. The echogenicity of testicular parenchyma increased in direct proportion to animal age, being higher in pubertal lambs when compared to prepubertal at the same age. The mediastinum testis was observed in 100% of the evaluated animals, regardless of the scan plane used, and was classified as diffuse or moderately or highly echogenic. Echogenicity and the thickness of the mediastinum testis increased in direct proportion to animal age. The epididymal tail was presented in hypoechoic relation to the testicular parenchyma. Based on these results, it was concluded that ultrasound is useful tool for selection and morphophysiological evaluation of Santa Inês lambs on peripubertal and pubertal phases, when used in combination with other methods such as semen evaluation.

Ultrassonografia testicular em caprinos jovens da raça Alpina / Testicular ultrasonography in young Alpine goats

Objetivou-se com este trabalho observar as mudanças do parênquima testicular de acordo com o fotoperíodo e com a faixa etária dos animais por meio da ecotextura testicular e a viabilidade do uso do ultrassom na avaliação testicular. Foram utilizados 4 animais desde o 6° até o 13° mês de idade. Os exames ecográficos foram realizados a cada 15 dias para avaliar a ecotextura testicular. O parênquima testicular mostrou-se moderadamente ecogênico e homogêneo. Foi observado comportamento semelhante na ecotextura testicular durante o período de avaliação, sendo maior no 8º mês de idade, quando o volume testicular também foi maior; tais mudanças foram relacionadas com o fotoperíodo da região. A avaliação ultrassonográfica dos testículos mostrou-se um método eficiente, capaz de detectar as mudanças no parênquima testicular ao longo do tempo, e como ferramenta alternativa para diferenciação de condições normais e patológicas em caprinos...

The purpose of this study was to observe changes in the testicular parenchyma of male goats. Changes were observed in terms of goat age, evaluating the echotexture and ultrasound viability in the testicles. Four animals were used, ranging from six to 13 months of age. The echographic examinations were performed every 15 days to assess echotexture of the testicle. The testicular parenchyma appeared moderately echogenic and homogeneous. Similar results were observed in testicular echotexture during the evaluation period. The highest results appearing during the eighth month of age, when the testicular volume was also highest; such changes were related to the regional photoperiod. The ultrasound evaluation of the testicles was shown to be an effective method, ultimately capable of detecting changes in testicular parenchyma over time, being an alternative to differentiation normal and pathological conditions in goats...

Escores de cloaca e de crista e morfometria testicular em galos de matriz pesada com 71 semanas de idade e três categorias de peso corporal / Cloaca and comb scores and testicular morphometry in broiler breeder roosters at 71 weeks of age and three body weight categories

Em galos de matriz pesada, o peso dos testículos se associa positivamente com a produção diária de espermatozoides e com a fertilidade. Contudo, os testículos de aves se localizam na cavidade abdominal, não sendo acessíveis para exame andrológico como os testículos de mamíferos. A avaliação reprodutiva de galos de matriz pesada é subjetiva e se baseia em características como peso corporal (PC), qualidade de pés e pernas e morfologia de cloaca e crista. Estudos que validem o potencial de associação dessas características fenotípicas com parâmetros da morfometria testicular são raros, principalmente quando se consideram galos de matriz pesada manejados em lotes comerciais. Escores de cloaca e de crista e parâmetros da morfometria testicular foram avaliados em galos Cobb 500 Slow adultos das categorias de PC leve (<4,300kg), médio (entre 4,300 e 5,250kg) e pesado (≥5,250kg). O coeficiente de variação para PC foi de 12%. Os galos da categoria de PC pesado apresentaram testículos mais pesados e maiores escores de cloaca e de crista do que os galos da categoria de PC leve. Não houve diferença estatística entre as categorias de PC, considerando-se índice gonadossomático, porcentagens de túbulo seminífero e de tecido intersticial, diâmetro de túbulo seminífero e altura de epitélio seminífero. Correlação positiva, moderada e significativa foi observada entre peso de testículo e características de PC, escores de cloaca e de crista, diâmetro de túbulo seminífero e altura de epitélio seminífero. Um dos seis galos da categoria de PC leve avaliados quanto à histologia de testículo apresentou atrofia testicular. Em galos Cobb 500 Slow, conclui-se que o PC influencia o peso e a qualidade histológica de testículos e que os escores de cloaca e de crista podem ser utilizados com potencial moderado de predição para identificar galos com baixo peso de testículos...

In broiler breeder roosters the weight of the testes is positively associated with daily sperm production and fertility. However, the avian testes are located in the abdominal cavity, not being accessible for reproductive evaluation as mammalian testes. The reproductive evaluation of broiler breeder roosters is subjective and is based on characteristics such as body weight (BW), quality of feet and legs and morphology of cloaca and comb. Studies that validate the potential of associating these phenotype characteristics with testicular morphometric parameters are rare, especially when broiler breeder roosters of commercial flocks are considered. Cloaca and comb scores and testicular morphometric parameters were evaluated in adult Cobb 500 Slow roosters with low (<4.300kg), medium (between 4.300 and 5.250kg) and high (≥5.250kg) BW categories. The coefficient of variation for BW was 12%. The high BW roosters have heavier testes and higher cloaca and comb scores than the low BW roosters. There was no statistical difference between the BW categories in gonadosomatic index, percentage of seminiferous tubule and interstitial tissue, diameter of the seminiferous tubule and height of the seminiferous epithelium. Testis weight is positive, moderate and significantly correlated with BW, cloaca and comb scores, diameter of seminiferous tubule and height of seminiferous epithelium. One of six low BW roosters evaluated for histology of the testis had testicular atrophy. We conclude that in Cobb 500 Slow roosters BW influences the weight and histological quality of the testes. Besides this, cloaca and comb scores can be used with moderate prediction potential to identify roosters with underweight testes...

Efecto de la maca peruana (lepidium meyenii) y de la melatonina sobre el desarrollo testicular del ratón expuesto a hipoxia hipobárica continua / Effect of peruvian maca (lepidium meyenii) and melatonin on testicular development of mice exposed to continuous hypobaric hypoxia

El estudio de la hipoxia hipobárica (HH) determina un problema de salud pública y laboral en poblaciones que habitan en zonas de altura. La disminución del oxígeno afecta a diferentes órganos, incluyendo el testículo. El organismo responde frente a la hipoxia estimulando la angiogénesis, el flujo sanguíneo testicular e incrementa la temperatura intraescrotal, lo cual produce un daño de la espermatogénesis. Nuestro estudio valoró el efecto que produce la HH sobre el testículo del ratón. Se utilizó una cámara hipobárica regulada a 4.200 metros sobre el nivel del mar (msnm), en periodos de hipoxia durante 8,3; 16,6 y 24,9 días, en comparación a un grupo control en normoxia (500 msnm). En estos tres grupos, a unos ratones se administró melatonina, a otros maca (Lepidium meyenii) y a otros la combinación de melatonina y maca. Los objetivos fueron evaluar si la ingesta de maca protege al testículo, reduciendo el daño generado por la hipoxia, y determinar un posible efecto sinérgico de la melatonina y de la maca. La exposición a HH continua produjo una disminución del diámetro de los túbulos seminíferos y del lumen tubular; además, el seminograma demostró una reducción del recuento espermático, un aumento de la teratozoospermia y una reducción de la calidad del ADN espermático. La administración de maca aislada o la combinación de maca y melatonina en animales sometidos a HH produjo una notable mejoría de los parámetros relacionados con la función de los espermatozoides, siendo significativos la disminución del número de espermatozoides con morfología anormal y de la compactación del DNA, alcanzando en algunos casos valores próximos a los de los animales normóxicos. Los datos del presente modelo de HH corroboran los excelentes beneficios que la ingesta de maca tiene sobre la capacidad reproductiva de poblaciones que viven en áreas geográficas de grandes alturas.

Hypobaric hypoxia (HH) is a decisive factor in human health in populations that reside at high altitude levels. Low oxygen rate affects most tissues and organs, including the testis. In humans, hypoxia stimulates angiogenesis, testicular blood flow and increases intrascrotal temperature which determines negative effects on sperm production. Our study researched the effects of HH in mice testicle. Mice were housed in a hypobaric chamber with a setting at 4,200 m above sea level during three different periods of hypoxia (8.3, 16.6 and 24.9 days). Control groups were housed at normoxic conditions (500 m above sea level). Hypoxic mice were treated with melatonin, maca plant (Lepidium meyenii) and melatonin and maca combination. The aim of present study was to determine if maca consumption protects testis against harmful effects of hypoxia and to determine a possible synergistic effect between melatonin and maca administration. In this article we have demonstrated that hypoxia produces a considerable decrease of seminiferous tubules diameter and lumen diameter. Moreover, seminogram showed a reduced sperm count, increased teratozoospermia and a reduction of DNA quality. The HH mice treatment with maca or maca-melatonin combination showed statistically significant improvement at sperm function parameters, and in the reduction of sperm morphology abnormalities and DNA compaction, in some cases attaining rates closer to those registered in normoxic mice. Our experimental data corroborates that maca consumption improves reproductive capacity of populations that inhabit high altitude regions.

Postnatal development of testis in gaddi goat (capra hircus) / Desarrollo postnatal del testículo en la cabra gaddi (capra hircus)

A study was conducted on the testicles of 30 male Gaddi goats, divided into 3 groups viz; prepubertal (0 day to 18 months), pubertal (<18 months to >5 yrs) and post- pubertal (>5 yrs) ages. The study revealed that the testis was covered by fibroserous tunica albuginia having outer fibrous layer and inner vascular layer with smooth muscle fibers at birth. The tunica albuginia continuously grew in thickness from prepubertal to postpubertal animals. The septula testis arising from the tunica albuginia, divided the parenchyma in lobules and converged at mediastinum testis. Few differentiated and undifferentiated Leydig cells were present in the intertubular spaces of neonates, which enormously increased in pubertal animals and replaced by fibroblasts in the postpubertal animals. The parenchyma comprised of solid sex cords in new born kids. These were converted into luminated tubules after 6 months of age and had clear cut tubuli contorti and tubuli recti leading to the mediastinum testis. In "0" day old kids, sex cords were filled with undifferentiated small cells located peripherally along the basement membrane and large primordial germ cells located centrally. By six months of age, latter started showing sign of degeneration and thus luminization of tubules started. Spermatogenesis startedin the seminiferous tubules of 12-18 months goat. In pubertal animals all the stages of spermatogenic cells and Sertoli cells were identified in the seminiferous tubules. The population of gonia cells and primary + secondary spermatocytes were 35% and 30% respectively. In postpubertal animals gonia cells were reduced to <30% and Primary + secondary spermatocytes were <20%. Many degenerating seminiferous tubules showed hyalinization, fibrosis and giant cells. The stroma and parenchyma ratio which was 35:65 at birth became 30:60 at 6 months, 25:75 at 12 months and 15:85 in pubertal and 20:80 in postpubertal animals. Increased stroma in last phase was associated with replacement of seminiuferous tubules by fibrous elements.

Se realizó un estudio en 30 testículos de cabras Gaddi machos divididos en 3 grupos: prepúberes (0 días a 18 meses), púberes (<18 meses y >5 años) y post-púberes (>5 años). El estudio reveló que los testículos al nacimiento estaban cubiertos por una túnica albugínea fibroserosa con una capa fibrosa externa y otra capa vascular interna con fibras musculares lisas. En los animales prepúberes hasta los pospúberes, la túnica albugínea mostró un crecimiento continuo de espesor. El septo testicular derivado de la túnica albugínea, divide el parénquima en lóbulos convergentes en el mediastino testicular. En los espacios intertubulares de los testículos de los recién nacidos, se observaron escasas células interticiales diferenciadas e indiferenciadas, sin embargo, se registró un aumento importante en los animales púberes las cuales fueron reemplazadas por fibroblastos después de la pubertad. El parénquima en los testículos de los animales recién nacidos estaba compuesto por cordones sexuales continuos. Estos se convierten en túbulos luminados después de los 6 meses de edad, evidenciando túbulos contorneados y rectos que conducen al mediastino testicular. En animales de 0 días de edad, los cordones sexuales presentaban pequeñas células no diferenciadas y situadas periféricamente a lo largo de la membrana basal y grandes células germinales primordiales céntricas. A los seis meses de edad, este último comenzó a mostrar signos de degeneración. La espermatogénesis se inició en los túbulos seminíferos desde los 12 a 18 meses de edad. En los animales púberes se identificaron en los túbulos seminíferos todas las etapas de las células de espermatogénesis y células sustentaculares. La población de células gonia fue del 35% y de espermatocitos primarios más secundarios 30%. En los animales pospúberes las células gonia se redujeron a <30% y espermatocitos primarios más secundarios a < 20%. Muchos túbulos seminíferos presentaron degeneración. La relación parénquima/estroma fue 35:65 al nacer, luego de 30:60 a los 6 meses, de 25:75 a los 12 meses, de 15:85 en animales prepúberes y 20:80 en la pubertad. El aumento del estroma en la última fase fue asociado con el reemplazo de los túbulos seminíferos por elementos fibrosos.

Desenvolvimento corporal e testicular em machos de codornas de corte e de postura de 25 a 360 dias / Body and testicular development in males of meat- and egg-type quails of 25 to 360 days

Codornas do tipo carne e tipo ovos foram analisadas para determinar o desenvolvimento reprodutivo, a puberdade e o início da senilidade. Para tal, 288 codornas (144 codornas de corte e 144 de postura) foram acompanhadas desde a eclosão até os 360 dias de idade. As aves foram distribuídas por idade em 18 grupos, sendo 8 codornas/grupo/tipo de codorna. Após 35 dias as codornas foram mantidas em condições de fotoperíodo de dias longos (17luz: 7escuro). O peso vivo e os valores morfométricos e histológicos testiculares foram determinados em cada período. Os dados obtidos foram analisados para determinar a curva de crescimento e o comportamento dos parâmetros analisados. O modelo que mais se adequou aos dados foi o modelo não linear de Gompertz (Y=A exp [-B e (-kt)]). O peso vivo e as características testiculares macro e microscópicas apresentaram comportamento alométrico entre si, sendo que, aproximadamente aos 60 dias os machos apresentaram-se sexualmente desenvolvidos, e estabilizaram o peso corporal por volta dos 100 dias. O testículo direito é mais cranial que o esquerdo e diferem em relação a comprimento e largura, porém não foi observada diferenças (P>0,05) para peso testicular. As codornas de corte apresentaram peso corporal e peso testicular maiores que as codornas de postura, porém as codornas de postura apresentaram peso relativo testicular maior. Durante todo o período analisado os machos puderam ser considerados sexualmente aptos. Os reprodutores apresentaram características sexuais ativas até os 360 dias de idade, representadas pelo tamanho testicular e pela atividade celular nos túbulos seminíferos.

Meat- and egg-type quails were analyzed to determine reproductive development, puberty and the onset of senility. For this purpose, 288 quails (144 meat- and 144 egg-type quails) were followed from hatch to 360 days old. The birds were distributed by age in 18 groups, 8 quails/group/type of quail. After 35 days quails were maintained under long daylength conditions (17L: 7D). The live weight and the macro and microscopical testicular characteristics were determined in each period. The data were analyzed to determine the growth curve and the behavior of parameters. The model that best fits the data was the non-linear Gompertz model (Y = A exp [-B and (-kt)]). Live weight and testicular morphometric parameters had an allometric behavior among themselves, and at approximately 60 days the males had sexually development, and body weight stabilized at about 100 days. The right testicle was more cranial than the left, differing in the length and width, but no difference in testicular weight (P>0.05) was observed. The meat quails had a larger body weight and testicular weight than the egg quails, but the egg-quail had a higher percentage of testes in relation to body weight. During the whole adult period the males could be considered sexually fit. Breeders had active sex characteristics until 360 days of age, represented by testicular size and by cellular activity in the seminiferous tubules.

Toxic effects of lead exposure during gestation and lactation on testicular development in albino rat offspring: a hormonal and histopathological study

This study aims at exploring the effect of lead exposure during pregnancy and lactation on the rat testis, which was evaluated via light and electron microscopic examination of testicular tissue, besides estimation of serum follicle stimulating hormone [FSH], luteinizing hormone [LH] and testosterone [TT] levels. Dams were intoxicated with 10 mg/kg body weight [BW]/day lead acetate throughout the gestational period and lactation. Pups were sacrificed on postnatal [PN] days 1 and 21. The weights of the body and testes, as well as serum gonadotropins and TT concentrations were significantly reduced in pups from lead-intoxicated mothers compared with their controls. Light microscopic examination of testicular samples from lead-exposed pups revealed thin-walled seminiferous tubules with significant reduction in their size. In addition, multiple vacuoles were seen within the tubules and in the interstitium, and the interstitial spaces appeared enlarged with reduced number of Leydig cells. Apoptotic bodies were found among the basal parts of the spermatogenic epithelium. Examination of testicular tissue from the same group by electron microscopy revealed more cellular details confirming the toxic effects of lead. The altered seminiferous tubules showed multiple cytoplasmic vacuoles and precipitates along the nuclear membranes in Sertoli cells. Several seminiferous tubules showed apoptotic cells with heterochromatic nuclei and dense cytoplasm, whereas other tubules appeared ensheathed by single layers of myoid cells. Leydig cells revealed cytoplasmic vacuolations and irregular nuclei with chromatin masses. The results of this study indicate that lead intoxication affected the normal development of germinal cells and disrupted the testicular structure and endocrinal functions

Stereological evaluation of the effects of momordica charantia, antioxidants and testosterone on seminiferous tubules of rat / Evaluación estereológica de los efectos de momordica charantia, antioxidantes y testosterona sobre los túbulos seminíferos de rata

Ubiquitous consumption of Momordica charantia is hinged on credence of its potency in alleviating ailments. Almost no data exist on its quantitative relationship to the testes. The aim was to study the histomorphometric effect of Momordica charantia seed extract (MC), ascorbic acid (C), alpha-tocopherol (E) and testosterone (T) on the testes of rats. Ninety five mature 6-8 week-old male Sprague-Dawley (S-D) rats allotted randomly into six groups. Group I: fed MC for 16 weeks. Group II: administered MC for 8 weeks, later distilled water for another 8 weeks. Group III: administered C, E, T and a combination of all three test solutions (CET) for 8 weeks and later fed MC for another 8 weeks. Group IV: received MC initially for 8 weeks and later administered C, E, T and CET for another 8 weeks. Group V: fed concurrently the extract, C, E, T and CET for 8 weeks. Group VI (control): administered C, E, T, CET and distilled water for 16 weeks. The doses for MC, C, E and T were 50 mg/100 g b.w/oral/day, 0.01, 20.0 and 0.05 mg/kg. The animals were sacrificed; testes estimated for volume. Diameter, cross sectional area (AC), number of profiles per unit area (NA), length density (LV) and numerical density (NV) of the seminiferous tubules determined. The mean testicular volumes, tubular diameters and AC showed significant decline (p 0.05) in Groups I and III compared to controls. On cessation of MC values recovered to baseline control value. Groups IV and V, were not significantly (p 0.05) different from controls. There was a significant (p 0.05) increase in NA, LV and NV of seminiferous tubules in Groups I and III compared to controls. Histomorphometric data supports a reversible deleterious effect of MC on testes.

El consumo ubicuo de Momordica charantia ha girado en torno a su potencial participación en el alivio de dolencias. El objetivo fue estudiar los efectos del extracto de la semilla de Momordica charantia (MC), ácido ascórbico (C), alfa-tocoferol (E) y testosterona (T) sobre parámetros histomorfométricos en testículos de ratas. Noventa y cinco ratas machos Sprague-Dawley (S-D) maduras (6-8 semanas de edad) fueron asignadas al azar en seis grupos. Grupo I: alimentados con MC durante 16 semanas. Grupo II: MC administrado durante 8 semanas, luego agua destilada por otras 8 semanas. Grupo III: C, E, T administrados junto a una combinación de las tres soluciones de prueba (CET) durante 8 semanas y luego alimentados con MC por otras 8 semanas. Grupo IV: MC inicialmente durante 8 semanas y posteriormente C, E, T y CET por otras 8 semanas. Grupo V: alimentado al mismo tiempo con el extracto, C, E, T y CET durante 8 semanas. Grupo VI (control): administrado con C, E, T, CET y agua destilada durante 16 semanas. Las dosis de MC, C, E y T fueron de 50 mg/100 g de peso corporal/oral/día, 0,01, 20,0 y 0,05 mg/kg. Los animales fueron sacrificados y se estimó el volumen de los testículos. El diámetro, área de sección transversal (AC), el número de perfiles por unidad de área (NA), la densidad de longitud (LV) y la densidad numérica (NV) de los túbulos seminíferos fueron determinados. Los volúmenes medio testicular, diámetro de túbulos y AC mostraron una disminución significativa (p 0,05) en los grupos I y III en comparación con los controles. Al quitar MC, los valores se recuperaron al valor basal de control. Los grupos IV y V, no fueron significativamente diferentes de los controles (p 0,05). Hubo una diferencia significativa (p 0,05) con aumento de la NA, LV y NV de los túbulos seminíferos de los grupos I y III en comparación con los controles. Los datos histomorfométricos apoyan la reversibilidad de los efectos nocivos de la MC en los testículos.

Propylthiouracil-induced hypothyroidism delays apoptosis during the first wave of spermatogenesis

Mammalian germ cell apoptosis plays a key role in controlling the correct number of germ cells supported by Sertoli cells during the first wave of spermatogenesis in mammalian puberty. However, little is known about hormonal factors that could influence the rate of germ cell apoptosis during puberty or adulthood. In this work we evaluate germ cell apoptosis under hypothyroidism induced by goitrogen propylthiouracil (PTU) during the first wave of spermatogenesis. Neonatally administered PTU promoted a delay in the differentiation of Sertoli cells as evaluated by the expression of clusterin using immunohistochemistry and RT-PCR. Clusterin had different expression levels in control and PTU-treated animals, but under both conditions the highest levels were found in 35-day-old rats. In addition, clusterin displayed a cytoplasmic localization in control testes, but appeared located in the nucleus in PTU-treated animals. The wave of apoptosis (determined by caspase activity and quantification of apoptotic cells) characteristic of the first round of spermatogenesis was delayed by at least 10 days in these animals. The expression levels of proapoptotic genes like BAX or BAD were different between control and PTU-treated rats; although in both groups the highest level was found at the same age (days). Thus our results indicate that the characteristic pubertal apoptotic wave during rat spermatogenesis is delayed in neonatal hypothyroid rats.