Efecto de la Vitamina E en Células Peritubulares de Testículos de Ratones Mus musculus Tratados con Ácido Valproico Durante el Estado Fetal y Pubertad / Effect of Vitamin E on Peritubular Cells from Testis of Mus musculus Mice Treated with Valproic Acid During the Fetal Stage and Puberty

El ácido valproico (VPA) es un fármaco antiepiléptico teratógenico que, al ser administrado durante etapas tempranas del embarazo, puede producir alteraciones en el desarrollo embriofetal, las que se manifiestan tanto a nivel del sistema nervioso como del testículo. No obstante, se ha reportado que la administración de vitamina E (VE) podría revertir dichas alteraciones. El objetivo del presente estudio fue determinar el efecto protector de la VE a nivel testicular en fetos y ratones púberes expuestos a VPA durante la fase embrionaria de su desarrollo. Se utilizó un total de 30 ratones hembra adultas gestantes (Mus musculus) cepa BALB/c, las cuales se dividieron en 6 grupos. El estudio contempló el análisis de fetos machos a los 17,5 días post-coital (dpc) y machos juveniles a las 6 semanas post-natal. A los grupos 1 y 4 se les administró 0,3 mL de solución fisiológica (grupos control para 17,5 dpc y 6 semanas postnatal, respectivamente). A los grupos 2 y 5 se les suministró la cantidad de 600 mg/kg de VPA (grupos VPA), en tanto que a los grupos 3 y 6 se les aplicó la misma dosis de VPA complementada con 200 UI de VE (grupos VPA+VE). Se describió la histología normal y patológica del compartimento peritubular del testículo. En los grupos VPA se evidenció una degeneración de la pared peritubular, y atrofia de túbulos seminíferos, así como exfoliación de las células germinales. Por el contrario, en los grupos VPA+VE tales signos no fueron observados y la morfología presentó aspecto normal solo con algunas alteraciones focales. Estos resultados corroboran el hecho que la administración de VE contrarresta en parte, los efectos deletéreos que ocasiona el VPA.

SUMMARY: Valproic acid (VPA) is a teratogenic antiepileptic drug that, when administered during the early stages of pregnancy, can produce alterations in embryo-fetal development, which manifest both at the level of the nervous system and the testicle. However, it has been reported that the administration of vitamin E (VE) could reverse these alterations. The study aimed to determine the protective effect of VE at the testicular level in fetuses and pubertal mice exposed to VPA during the embryonic phase of their development. 30 pregnant adult female mice (Mus musculus) BALB/c strain were used, which were divided into 6 groups. The study included the analysis of male fetuses at 17.5 days post-coital (dpc) and juvenile males at 6 weeks post-natal. Groups 1 and 4 were administered 0.3 mL of physiological solution. Groups 2 and 5 were given 600 mg/kg of VPA (VPA groups), while groups 3 and 6 were given the same dose of VPA supplemented with 200 IU of VE (VPA+VE). The normal and pathological histology of the peritubular compartment of the testis was described. In the VPA groups, degeneration of the peritubular wall, and atrophy of the seminiferous tubules, as well as exfoliation of the germ cells, were evident. On the contrary, in the VPA+VE groups such signs were not observed and the morphology presented a normal appearance with only some focal alterations. These results corroborate the fact that the administration of VE partially counteracts the deleterious effects caused by VPA.

Characteristics of the puberty in hair ram lambs and its crosses in Colombia under low altitude conditions / Características de la pubertad en corderos de pelo y sus cruces en Colombia en condiciones de baja altitud

ABSTRACT Objective. Aimed to describe the characteristics of the onset of puberty in males in Colombian hair ram lambs (Ovino de Pelo Colombiano, OPC), and their crosses with Katahdin and Santa Inés sheep in a farm located in Villavicencio, Meta. Materials and methods. 15 lambs of three biotypes: OPC x OPC (OPC), Santa Ines x OPC (SO) and Katahdin X OPC (KO) from four until 12 months old. Ram lambs were grazing and they had supplementation with commercial salt and water ad libitum. Monthly body weight (BW), scrotal circumference (SC), testicular volume (TV) were measured and testosterone level were determined by Elisa test, and ultrasound of the two testicles was performed to determine the presence of the testicular mediastinum, also evaluating the presence and detachment of the urethral prolongation and then electro ejaculation was performed to determine the macroscopic and microscopic characteristics of the semen. Results. Genotype effect was significant for evolution of body BW, SC, TV over time. The SO and KO crosses presented the highest values in BW, whereas OPC lambs maintained a lower value until the end of the assay. At six months old, the three biotypes presented a minimum concentration of 150 million of sperm per ml with 30% of individual progressive motility. Conclusions. In non-seasonal tropical conditions in Colombia (Orinoquia), depending on the variables included, body weight, testicular development, pennis morphology, semen quality, sperm concentration and testosterone levels, it is postulated that around six months of age, the onset of puberty is displayed in the three biotypes.

RESUMEN Objetivo. Describir las características del inicio de la pubertad en corderos machos (Ovino de Pelo Colombiano, OPC) y sus cruces en una granja ubicada en Villavicencio, Meta. Materiales y métodos. Se evaluaron 15 corderos de tres razas: OPC x OPC (OPC), Santa Inés x OPC (SO) y Katahdin X OPC (KO), a partir de los cuatro meses de edad, cinco corderos por cruzamiento, manejados en pastoreo rotacional, con sal mineralizada comercial y agua a voluntad. Mensualmente hasta los doce meses de edad, se evaluó el peso corporal (PC), circunferencia escrotal (CE), volumen testicular (VT), y se determinó la concentración de testosterona en suero mediante la prueba de Elisa, se determinó la presencia del mediastino testicular mediante ecografía y se evalúo el desprendimiento de la prolongación uretral. Finalmente se determinaron las características macroscópicas y microscópicas del eyaculado. Resultados. El efecto racial o de cruzamiento fue significativo para los resultados del PC, CE y VT en el tiempo. Los cruces OPC y KO presentaron los valores más altos en PC, mientras que los corderos OPC mantuvieron un valor menor de PC hasta el final de la investigación. Se realizaron al menos cuatro evaluaciones seminales hasta la presentación de una concentración mínima de 150 millones de espermatozoides por ml con un 30% de motilidad progresiva individual. Conclusiones. En condiciones tropicales no estacionales en Colombia (Orinoquia), para las variables peso corporal, desarrollo testicular, morfología del pene, calidad del semen, concentración de espermatozoides y niveles de testosterona, se postula que alrededor de los seis meses de edad, se presenta el inicio de la pubertad en los tres biotipos.

PI3K-Akt/LKB1-AMPK-mTOR-p70S6K/4EBP1 signaling pathways participate in the regulation of testis development and spermatogenesis: An update / 中华男科学杂志

Male infertility is closely associated with spermatogenesis disorders triggered by aberrant gene expression or abnormal signaling pathways in the testis. The mammalian target of rapamycin (mTOR) is a central regulator of cell metabolism, playing an important role in regulating cell proliferation, differentiation, translation, actin polymerization, cycle progression, energy metabolism, autophagy, and other cellular activities. PI3K-Akt and LKB1-AMPK, the two well-defined classic signal transduction pathways, regulate the expressions of mTOR and its downstream p70S6K/4EBP1 through different molecular pathways. Recent studies show that mTOR-p70S6K/4EBP1 signaling participates in the regulation of the proliferation and differentiation of testicular cells and spermatogenesis. This review focuses on the role of PI3K-Akt/LKB1- AMPK-mTOR signaling cascades in testis development and spermatogenesis, providing some new perspectives for the studies of the molecular mechanism underlying male sterility.

Desenvolvimento testicular e puberdade em machosda raça guzerá da desmama aos 36 meses de idadecriados no cerrado mineiro / Testicular Development and Puberty in Guzerat Males from Weaning to 36 Months of Age Raised at the Sierra Minera

O objetivo deste estudo foi caracterizar o desenvolvimento testicular, avaliar a associaçãoexistente entre o perímetro escrotal (PE) e o volume testicular (VOL), e identificar os parâmetrostesticulares na puberdade, em machos da raça guzerá. Foram avaliados 330 machostotalizando 1757 observações realizadas a cada três meses a partir da desmama até os 36meses. Foram feitas as mensurações do PE, comprimento e largura testiculares. O VOL foicalculado seguindo método descrito por Fields et al. Os animais que apresentaram PE ≥ 20 cmforam submetidos à coleta de sêmen utilizando-se estímulo eletro-ejaculatório. Foram consideradosanimais púberes aqueles que apresentaram pelo menos um espermatozoide móvelno ejaculado. A curva de desenvolvimento testicular foi descrita pela função logística, sendoo ponto de inflexão máximo do perímetro escrotal e do volume testicular aos 13,2 meses deidade (18,1 cm) e 23,3 meses de idade (389,4 cm3), respectivamente. A taxa de crescimentomédia do perímetro escrotal e do volume testicular antes e depois do ponto de inflexãoforam respectivamente: 0,58 cm/m, 16,3 cm3/m, 0,29 cm/m, 7,7 cm3/meses de idade. Aidade média na puberdade foi de 19,6 meses, com 250,6 kg de peso, 22,8 cm de perímetroescrotal e 284,7 cm3 do volume testicular. A proporção de machos púberes na faixa etária de12-16 meses foi de 7,4 %; dos 16-20 meses foi 41,23 %; dos 20-24 foi 68 %; dos 24-28 foi91,96 %. Aos 24 meses de idade, 91,96 % dos animais encontravam-se púberes. Foi verificadauma alta correlação positiva entre PE e VOL (r = 0,91; P<0,001). Estes resultados indicamque na raça guzerá o perímetro escrotal estima de forma eficiente o volume escrotal, podendoser usado como uma medida segura para a seleção de jovens reprodutores...

El presente estudio caracterizó el desarrollo testicular, evaluó la asociación existente entre el perímetro escrotal (PE) y el volumen testicular (VOL) e identificó los parámetros testiculares en la pubertad en machos de la raza guzerat. Se evaluaron 330 machos que totalizaron 1757 observaciones realizadas cada tres meses, desde el destete hasta los 36 meses. Serealizaron mediciones del PE, longitud y ancho testiculares. Se calculó el VOL siguiendo el método descrito por Fields et ál. Los animales que presentaron PE ≥ 20 cm fueron sometidos a la recolección de semen utilizando estímulo electroeyaculatorio. Se consideraron púberes los animales que presentaron por lo menos un espermatozoide móvil en el eyaculado. La curva de desarrollo testicular se describió utilizando la función logística, siendo el punto de inflexión máximo del perímetro escrotal y del volumen testicular a los 13,2 meses de edad (18,1 cm) y 23,3 meses de edad (389,4 cm3), respectivamente. La tasa de crecimiento promedio del perímetro escrotal y volumen testicular antes y después del punto de inflexión fueron respectivamente 0,58 cm/m; 16,3 cm3/m; 0,29 cm/m; 7,7 cm3/meses de edad. La edad promedio en la pubertad fue de 19,6 meses, con 250,6 kg de peso, 22,8 cm de perímetro escrotal y 284,7 cm3 de volumen testicular. La proporción de machos púberes en la franja de12-16 meses fue de 7,4 %; de 16-20 meses, 41,23 %; de 20-24, 68 %; de 24-28 fue de 91,96 %.A los 24 meses de edad 91,96 % de los animales se encontraban púberes. Se verificó una altacorrelación positiva entre PE y VOL (r = 0,91; P < 0,001). Estos resultados indican que en la raza guzerat el perímetro escrotal predice de forma eficiente el volumen escrotal, pudiendo ser usado como una medida segura para la selección de jóvenes reproductores...

The objectives of this study were to characterize testicular development, assess the existingassociation between scrotal circumference (SC) and testicular volume (VOL), as well as toidentify testicular parameters in puberty of males of the Guzerat breed. Three hundred thirtymales were evaluated, totaling 1757 observations every three months, from weaning to 36months of age. SC, testicular length and width were measured. VOL was calculated accordingto the method described by Fields et al. Animals with SC ≥ 20 cm were subjected tosemen collection through electro-ejaculation stimulation. Animals with at least one motilesperm in the ejaculate were considered as pubertal. The testicular development curve wasdescribed using the logistic function, where the maximum inflection point of scrotal circumferenceand testicular volume was at 13.2 months (18.1 cm) and 23.3 months (389.4cm3), respectively. The average growth rate of the scrotal perimeter and testicular volume beforeand after the inflection point was 0.58 cm/m, 16.3 cm3/m, 0.29 cm/m, 7.7 cm3/monthsof age, respectively. Average age in puberty was 19.6 months, with 250.6 kg of weight, 22.8cm of scrotal perimeter and 284.7 cm3 of testicular volume. The proportion of pubescentmales at 12-16 months was 7.4 %; 41.23 % at 16-20 months; 68 % at 20-24 months; 91.96 %at 24-28 months. At 24 months of age, 91.96 % of the animals were pubescent. A highpositive correlation was found between SC and VOL (r = 0.91; P<0.001). These resultsindicate that, in the Guzeratel race, scrotal circumference efficiently predicts scrotal volumeand can therefore be used as a reliable measure for selecting young reproducers...

Identification of a Novel Gene by EST Clustering and its Expression in Mouse Ovary and Testis / 대한불임학회지

OBJECTIVE: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-5-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. METHODS: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named 5-day-ovary-specific gene-1 (5DOS1) and submitted to GenBank (accession number AY751521). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. RESULTS: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. CONCLUSIONS: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.