Androgen and oestrogen receptors in the growing antlers velvet of adult and yearling pampas deer (Ozotoceros bezoarticus) males.

To determine the presence, quantity and distribution of androgen (AR) and oestrogen receptors (ER) in the antler velvet (AV), samples were collected from adult and yearling pampas deer males, as well as skin from the ventrolateral abdominal area (ASK). Samples were analysed with ligand-binding assays for AR and ER activity and processed for AR and ERß immunohistochemistry. There was more content of AR in adults than in yearling males (p = 0.02), as well as a category and type of tissue interaction (p = 0.03). There was more ER content in adults than in yearling males (p = 0.005) and in the AV than in the skin (p = 0.0001). The AR-positive score (PS) was greater in AV than in ASK in the surface stroma (p = 0.0007). In the intermediate epidermis, the AR PS was greater in adults than in yearling males (p = 0.04) and in the ASK than in the AV (p < 0.0001). There was a male category and type of tissue interaction for AR PS in the sebaceous glands (p = 0.014). The ERß PS in the surface stroma was greater in ASK than in AV (p = 0.004) and tended to be greater in yearling than in adult males (p = 0.093). The ERß PS in the intermediate epithelium and the sebaceous glands was greater in adults than in yearlings (p = 0.004 and p = 0.007, respectively). In conclusion, we reported for the first time the presence of AR and ER in the velvet skin of growing antlers in pampas deer males. Therefore, the velvet skin is sensitive to both androgens and oestrogens. Furthermore, the greater content of oestrogen receptors in the velvet of adult males suggests that adults are more sensitive to this hormone than yearlings, and thus, oestrogens have greater importance in velvet activity regulation in adult males.

Collagen, glycosaminoglycans and matrix metalloproteinase-2 and metalloproteinase-9 in the cervix of the ewe during prepubertal development.

The tortuous nature of the ovine cervix restricts the transcervical passage of the cannula, and many studies have aimed to understand the endocrine mechanism of the remodelling of cervical tissue in adult ewe. However, little is known about the remodelling of the cervical tissue during the prepubertal development of the lambs. To obtain histochemical and biochemical evidence about the nature of the prepubertal development of the cervix of the ewe, cervices of Corriedale lambs obtained at 0, 1, 2, 4, 6 and 8 months of age (n = 5 to 6 in each) were processed. Neutral and acidic glycosaminoglycans (by PAS-Alcian stain) were weakly in the cervical stroma and not shown change during the development, whereas the percentage volume of fibrillar collagen (by van Gieson stain) increases throughout the experimental period in the superficial fold stroma and deep wall stroma (p < 0.05). The relative cervical weight (g/kg of body weight) and the collagen concentration (by spectrophotometry, mg/mg wet tissue) showed an early decreasing phase from months 0 to 4 and a later increasing phase from months 4 to 8 (p < 0.05). The latent form of matrix metalloproteinase-2 (MMP-2) detected by gelatin zymography (ng/mg protein) decreased from months 0 to 2 and increased from months 4 to 8, whereas the activated form decreased from months 0 to 2, remained low until month 6 and then recovered on month 8 (p < 0.0001). Data suggest that the relative cervical weight biphasic pattern during the development is related to MMP-2-dependent changes in the collagen content.

"Subnutrition effects during pregnancy and lactation on mitosis, apoptosis and androgen receptor expression in the rat testis".

In order to study the effect of undernutrition during gestation on the testicular development in rats and its impact on mitosis, apoptosis and the relative abundance of androgen receptor expression, twenty primiparous 3-month-old Sprague-Dawley (Rattus norvegicus) rats, weighing 246 ± 4.0 grams when the experiment began, were mated by the same male. Control group (CG), n = 10, fed ad libitum with water and rat chow and restricted group (RG, n = 10) fed throughout pregnancy and until birth with 40% of the ad libitum maternal daily feed intake. Litters from both groups suckled for 25 days, RG with 14 pups/litter and CG with 8 pups/litter. After weaning, all animals had access to ad libitum food and water. Testicular samples were taken from male pups at 2, 25 and 100 days of age. Immunohistochemistry was used to evidence androgen receptor (AR) expression in apoptotic (caspase 3-positive) and proliferating (PCNA-positive) cells. Three hundred nuclei of sustentacular (or Sertoli: SC), interstitial (or Leydig: LC), myoid (MC) cells as well as gonocytes (GC) were evaluated. Neither LC nor GC showed any differences between groups. However, SC androgen receptor (AR) positivity index in neonatal animals was lower in RG (1.27 ± 0.22 vs. 1.65 ± 0.17**). MC showed lower AR positivity index at 2 (2.69 ± 0.046 vs. 2.8 ± 0.055**) and 25 (1.34 ± 0.097 vs. 1.56 ± 0.1***) days of life; at 100 days of life, there was a greater number of apoptotic MC in the RG (8.5 ± 0.4 vs. 2.95 ± 1.1***). Thus, the present experiment demonstrates that the population dynamics of MC are affected by foetal programming due to undernutrition.

La reserpina aumenta la expresión de BDNF y PCNA, y disminuye la de caspasa-3, en células intersticiales (células de Leydig) de ratas / Reserpine increases BDNF and PCNA expression, but decreases caspase-3, in rat interstitial cells (Leydig cells)

La reserpina es un antipsicótico e hipotensor arterial que reduce significativamente los niveles de monoaminas centrales, y también es utilizada para modelar los cuadros depresivos humanos en animales de laboratorio. Este trabajo estudió, en ratas Wistar machos adolescentes, cómo la reserpina afecta indicadores moleculares de la función testicular, la cual se ha visto alterada en humanos deprimidos. Una semana luego de finalizado el tratamiento con reserpina (4 dosis de 0,0 o 1,0 mg/Kg, cada 2 días) la respuesta ansiosa y depresiva fue evaluada en un laberinto en cruz elevado. Posteriormente, se sacrificaron los animales y disecaron los testículos, los cuales fueron fijados e incluidos en bloques de parafina de donde se obtuvieron cortes histológicos de 6 µm de espesor. Estos se utilizaron para medir el diámetro de los túbulos seminíferos y para medir por inmunohistoquímica el porcentaje de células intersticiales (células de Leydig) positivas a (1) Factor neurotrófico derivado del cerebro, (2) antígeno nuclear de células en proliferación (BDNF y PCNA, respectivamente, por sus siglas en inglés), y a (3) caspasa-3. Se obtuvo también un índice de positividad al receptor de andrógenos en las células intersticiales. La expresión del receptor de andrógeno fue evaluada utilizando una escala semicuantitativa de escores (0, 1, 2 y 3) y el resto de las moléculas por presencia o ausencia de expresión de cada antígeno investigado en 300 células por preparado. Los resultados comportamentales indicaron alteraciones en la respuesta de ansiedad y una significativa depresión motora (e.g., mayor latencia en conductas de escape del sector blanco) en los animales tratados con reserpina. No se observaron diferencias en los diámetros de los túbulos seminíferos ni en la expresión del receptor de andrógeno, mientras que sí se encontró mayor proporción de células intersticiales positivas a BDNF y PCNA, y menor proporción de células positivas a caspasa-3, en los animales tratados. Los resultados corroboran la capacidad de la reserpina para reproducir rasgos comportamentales de la depresión. La administración de la droga, sin embargo, no parece reproducir a nivel testicular los efectos deletéreos encontrados en humanos deprimidos, e incluso los resultados sugieren que la reserpina puede mejorar algunos aspectos de la funcionalidad testicular relacionadas con la actividad de las células intersticiales en ratas.

Reserpine, a drug that depletes central monoamines, has been used as an antipsychotic and arterial hypotensive, and to model depression in animals. The present study analyzed, in adolescent male rats, the effects of chronic reserpine treatment on molecular indexes of testicular function. A week after termination of the treatment (4 doses of 0,0 or 1,0 mg/Kg/every 48 h) the animals were tested for anxiety response and depression patterns in an elevated plus maze. They were then euthanized, their testes dissected, fixed and embedded in paraffin to obtain blocks. Histological sections (6 µm) were obtained and used to measure the diameter of seminiferous tubules and the expression in Leydig cells of Brain-derived neurotrophic factor (BDNF), Proliferating cell nuclear antigen (PCNA), Caspase-3 and androgen receptors, by immunohistochemistry. Behavioral results indicated significant alterations in anxiety responses and a significant motor depression (e.g., greater latency to escape from the white sector). There were no differences between groups in the diameter of seminiferous tubules nor in the androgen receptors positivity. Reserpine-treated animals, however, exhibited more BDNF and PCNA positive cells, and less positive Caspase-3 cells in Leydig cells, than control animals. The results corroborate the efficacy of reserpine to reproduce some of the behavioral components of depression. The drug, however, does not seem to exert in rats the same effects on testicular function that have been found in humans diagnosed with depression. Furthermore the drug seems to enhance some aspects of testicular function related to Leydig cells function in rats.

Amphetamine, but not methylphenidate, increases ethanol intake in adolescent male, but not in female, rats.

Introduction: There has been an increasing interest in analyzing the interactions between stimulants and ethanol during childhood and adolescence. Stimulants are used to treat attention-deficit hyperactivity disorder (ADHD) in these developmental stages, during which ethanol initiation and escalation often occur. Methods: This study assessed the effects of repeated d-amphetamine (AMPH) or methylphenidate (MPH) treatment during adolescence [male and female Wistar rats, between postnatal day (PD) 28 to PD34, approximately] on the initiation of ethanol intake during a later section of adolescence (PD35 to PD40). Results: Amphetamine and MPH exerted reliable acute motor stimulant effects, but there was no indication of sensitized motor or anxiety responses. MPH did not affect dopamine (DA) levels, whereas AMPH significantly reduced insular levels of DA in both sexes and norepinephrine levels in females only. Repeated treatment with AMPH, but not with MPH, enhanced ethanol intake during late adolescence in male, but not in female, rats. Conclusion: A short treatment with AMPH during adolescence significantly altered DA levels in the insula, both in male and females, and significantly enhanced ethanol intake in males. The present results suggest that, in adolescent males, a very brief history of AMPH exposure can facilitate the initiation of ethanol intake.