Alterations of spermatogenesis in etoposide-treated rats: A stereological study

The etoposide is an anticancer drug that interacts with topoisomerase II. Thirty-day-old rats received intraperitonially 2mg/kg of etoposide for 30 consecutive days. Their testes were analyzed in the adult phase under light microscopy according to histomorphometric and stereological parameters. Random 3mum-thick-paraplast sections of testis were stained with periodic acid-Schiff reaction and Harris' hematoxylin method. Serum testosterone level and reproductive performance were also investigated. The results showed an accentuated decrease in the frequency of germinal lineage cell types and differentiated spermatogonia were the most affected cell types. Morphometric and stereological testicular parameters exhibited highly, significant reductions in adult etoposide-treated rats. Their reproductive performance diminished but their serum testosterone level was not significantly altered. The mortality frequency of the progenies was 100%.

CHEMICAL SYMPATHECTOMY BLOCKS ANDROGEN BIOSYNTHESIS DURING PREPUBERTY

Twenty-one-day old male Wistar rats were injected subcutaneously with guanethidine (GUA) at doses of 5 and 10 mg kg(-1) day(-1) for 20 days. Animals were sacrificed by decapitation during the prepubertal (41 days of age) and early-pubertal (51 days of age) periods of sexual development. The testes were collected, frozen in liquid N-2 and stored at -70 degrees C until determination of testicular progesterone (P): androstenedione (A) and testosterone (T). Higher levels of P (2.18 +/- 0.24 ng/g. control = 1.24 +/- 0.16 ng/g) associated with decreased levels of androgens (A = 0.26 +/- 0.06 ng/g and T = 2.05 +/- 0.19 ng/g; control = 1.86 +/- 0.76 ng/g and 8.48 +/- 1.16 ng/g, respectively) were observed in 10 mg GUA-treated rats of prepubertal age, while only P levels (3.12 +/- 0.51 ng/g control = 1.73 +/- 0.27 ng/g) were increased in rats of early pubertal age. It is important to note that in 41-day old male rats both 5 and 10 mg were effective in decreasing testicular concentration of testosterone. These results suggest that the sympathetic innervation of the testis is involved in the modulation of androgen biosynthesis, acting through a selective step in the steroid biochemical pathway during the pubertal process and that under the conditions employed the blockage in androgen biosynthesis in the prepubertal stage of sexual maturation is dependent on the dose of GUA.

Seasonal morphology of the domestic quail (Coturnix coturnix japonica) testis

A histological study was conducted on the testes of adult domestic quails (Coturnix coturnix japonica) over a year. The results revealed a clear variation of testicular weight, seminiferous tubule diameter, and the thickness and composition of the germinal epithelium over the year. The highest testicular weights were detected at the end of the autumn and during the winter (short-day period), reaching a maximum, together with spermatogenic activity, in September (long-day period in the southern hemisphere). In contrast, both testicular weight and spermatogenic activity were markedly decreased at the end of spring and during summer (long-day periods in the southern hemisphere).

Amifostine protective effect on cisplatin-treated rat testis

Cisplatin is a potent drug used in clinical oncology but causes spermatogenesis damage. Amifostine is a drug used against toxicity caused by ionizing irradiation and chemotherapeutic drugs. Since cisplatin provokes fertility and induces germ cell apoptosis and necrosis, we proposed to evaluate the amifostine cytoprotective action on testes of cisplatin-treated rats. Thirty-day-old prepubertal Wistar rats received a single cisplatin dose of 5 mg/kg and were killed after 3, 6, and 12 hr. The hematoxylin-eosin stained testicular sections were submitted to histological, morphometric, and stereological analysis. The terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling (TUNEL) method was used to label apoptotic cells. TUNEL-positive and TUNEL-negative germ cells with abnormal nuclear morphology (ANM) were scored. Significant alterations of greater part of the parameters occurred in the cisplatin-treated group (CE) compared to the group that received amifostine before the cisplatin-treatment (ACE); however, testicular weight and volume did not vary between these groups. Tubular diameter was reduced in CE in comparison to ACE rats, while interstitial tissue and lymphatic space volume and volume density were significantly higher in CE rats; interstitial testicular edema probably occurred in cisplatin-treated rats. CE rats showed important histological alterations, which were more accentuated than in ACE rats. The numerical densities of apoptotic germ cells and TUNEL-negative cells with ANM were lower in ACE than in CE rats. In conclusion, the amifostine previously administered to prepubertal rats reduced the testicular damage caused by cisplatin. We conclude that amifostine partially protected the rat seminiferous epithelium against cisplatin toxicity.

Heritability Estimate and Genetic Correlations of Reproductive Features in Nellore Bulls, Offspring of Super Precocious, Precocious and Normal Cows Under Extensive Farming Conditions

The present work aimed to estimate heritability and genetic correlations of reproductive features of Nellore bulls, offspring of mothers classified as superprecocious (M1), precocious (M2) and normal (M3). Twenty one thousand hundred and eighty-six animals with average age of 21.29 months were used, evaluated through the breeding soundness evaluation from 1999 to 2008. The breeding soundness features included physical semen evaluation (progressive sperm motility and sperm vigour), semen morphology (major, minor and total sperm defects), scrotal circumference (SC), testicular volume (TV) and SC at 18 months of age (SC18). The components of variance, heritability and genetic correlations for and between the features were estimated simultaneously by restricted maximum likelihood, with the use of the vce software system vs 6. The heritability estimates were high for SC18, SC and TV (0.43, 0.63 and 0.54; 0.45, 0.45 and 0.44; 0.42, 0.45 and 0.41, respectively for the categories of mothers M1, M2 and M3) and low for physical and morphological semen aspects. The genetic correlations between SC18 and SC were high, as well as between these variables with TV. High and positive genetic correlations were recorded among SC18, SC and TV with the physical aspects of the semen, although no favourable association was verified with the morphological aspects, for the three categories of mothers. It can be concluded that the mothers sexual precocity did not affect the heritability of their offspring reproduction features.

A comparative study of the extracutaneous pigmentary system in three anuran amphibian species evaluated during the breeding season.

The present investigation examined the extracutaneous pigmentation pattern of three species of anuran amphibians, Dendropsophus nanus, Physalaemus cuvieri, and Rhinella schneideri, during the course of their breeding seasons. Pigmentation intensity in the different organs was graded on a 4-category scale, in which category 0 refers to organs without pigment and category 3 refers to intensely pigmented organs, with 2 intermediate stages of progressively stronger pigmentation. Rhinella schneideri showed testicular pigmentation, with intra-specific variation (categories 0 and 1). In P. cuvieri the pigmentation in the lungs varied in time: all the animals showed pigmentation (category 1) at the beginning of the breeding season, and as the season progressed the absence of pigments became the most common pattern. In the liver of the first animals collected, the pigment intensity was high (category 2) with many iridophores present, but in the last specimens collected no iridophores were found. The variation in D. nanus occurred in the kidneys, where animals collected at the beginning of the season did not show pigmentation. Renal veins displayed few melanocytes (category 1); animals collected at the end of the season showed more pigmentation in the kidneys (category 2), whereas in the renal veins the intensity remained the same. The changes observed in the extracutaneous pigment system in some organs, during the reproductive period, may be due to physiological alterations or may represent a species-specific characteristic.

Effects of gonadectomy on prolactin and LH secretion and the pituitary-thyroid axis in male dogs

The effects of gonadectomy on the secretion of prolactin, LH, TSH, and thyroxine were investigated. Blood serum hormone concentrations were analysed before and at 20, 120, and 180 min after a single iv TRH injection in each of eight healthy intact and castrated male beagle dogs before (control) and after 4-week treatment with the dopamine-2 receptor agonist cabergoline. Under control conditions the mean prolactin, TSH, and thyroxine concentrations were similar in intact and gonadectomised dogs, and administration of TRH provoked a significant (p < 0.01) increase in concentrations of the three hormones. The overall inhibitory effect of cabergoline treatment on prolactin secretion was more pronounced in the castrated dogs compared with the intact group. Cabergoline significantly suppressed the TRH-induced prolactin increase in each group (p < 0.01). Corresponding TRH-stimulated TSH concentrations were not affected by cabergoline. In the gonadectomised dogs, thyroxine concentrations before and at 120 and 180 min after TRH injection were significantly lower than under control conditions. LH concentrations were always higher (p < 0.01) in gonadectomised dogs compared with the intact dogs, but appeared to be affected neither by TRH nor by cabergoline administration. It can thus be concluded from the results, that gonadectomy does not result in hyperprolactinaemia in male dogs, while LH concentrations are significantly increased due to missing androgen feedback. Thyroid function remains unaffected by gonadectomy. Testicular steroids appear to interact with central dopaminergic and probably other neuroendocrine mechanisms regulating the secretion of prolactin, TSH, and thyroxine. Thus, long-term dopamine-2 receptor agonistic treatment may lead to a hypothyroid condition in castrated male dogs. (c) 2009 Elsevier B.V. All rights reserved.

Early Development and Putative Primordial Germ Cells Characterization in Dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reproductive biology of the neotropical rattlesnake Crotalus durissus from northeastern Brazil: a test of phylogenetic conservatism of reproductive patterns

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enhanced ERbeta immunoexpression and apoptosis in the germ cells of cimetidine-treated rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Selective Ablation of the Androgen Receptor in Mouse Sertoli Cells Affects Sertoli Cell Maturation, Barrier Formation and Cytoskeletal Development

The observation that mice with a selective ablation of the androgen receptor (AR) in Sertoli cells (SC) (SCARKO mice) display a complete block in meiosis supports the contention that SC play a pivotal role in the control of germ cell development by androgens. To delineate the physiological and molecular mechanism responsible for this control, we compared tubular development in pubertal SCARKO mice and littermate controls. Particular attention was paid to differences in SC maturation, SC barrier formation and cytoskeletal organization and to the molecular mediators potentially involved. Functional analysis of SC barrier development by hypertonic perfusion and lanthanum permeation techniques and immunohistochemical analysis of junction formation showed that SCARKO mice still attempt to produce a barrier separating basal and adluminal compartment but that barrier formation is delayed and defective. Defective barrier formation was accompanied by disturbances in SC nuclear maturation (immature shape, absence of prominent, tripartite nucleoli) and SC polarization (aberrant positioning of SC nuclei and cytoskeletal elements such as vimentin). Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35. Differences in the expression of SC genes known to play a role in junction formation could be shown from day 8 for Cldn11, from day 15 for Cldn3 and Espn, from day 20 for Cdh2 and Jam3 and from day 35 for ZO-1. Marked differences were also noted in the transcript levels of several genes that are also related to cell adhesion and cytoskeletal dynamics but that have not yet been studied in SC (Actn3, Ank3, Anxa9, Scin, Emb, Mpzl2). It is concluded that absence of a functional AR in SC impedes the remodeling of testicular tubules expected at the onset of spermatogenesis and interferes with the creation of the specific environment needed for germ cell development.

Anatomy of the male reproductive system of Phrynops geoffroanus (Testudines: Chelidae).

The reproductive system of male Phrynops geoffroanus adults is macroscopically described and the variation in testicular biometry is evaluated. A pair of oval testes is connected by the efferent ductules to the epididymis, which continue as deferent ducts, which emerge in the penis. The volume of the gonads showed the highest averages during spring and summer months. GSI varied significantly throughout the year, with the highest averages observed in the months that correspond to the end of spring and the beginning of summer, when the reproduction of the species takes place, and the lowest averages were seen in winter, suggesting a cyclical testicular activity.

Effect of alcohol on the rat juxtaprostatic pelvic ganglia and gonads.

In the present paper, it was investigated the aspect of the intrapelvic autonomic neuronal system (juxtaprostatic ganglion) and gonads of rats submitted to a experimental alcoholism. The animals which had water substituted for 35% sugar cane spirit for 30 d, presented testicular lesions as well as numeric and degenerative alterations in the juxtaprostatic pelvic ganglia neurons.

Características histoquímicas e estruturais do testículo do morcego hematófago (Desmodus rotundus rotundus, Geoffrey, 1810).

The testicular stroma of the vampire bat including the testicular capsula and the lamina propria of the seminiferous tubuli, was strongly PAS-positive. This observation was a possible indication of great amounts of structural glycogen and other glycoconjugates at the level of smooth muscle cells; elongated contractile cells and/or collagen frameworks of the tunica albuginea and tubular lamina propria. In the last the basement membranes of the seminiferous tubules were particularly strongly PAS positive, as an indication of their neutral mucosubstances structural composition, previously described (Malmi et al., 1987). The epithelium lining from the cavitary and surface rete testis complex showed low reactivities to mucosubstances; total proteins and lipids and oxidative enzymes studied. Although the apical granulation at the rete testis epithelium showed an intense PAS reactivity with hypothesis of glycoprotein secretion, through the rete. The PAS, Sudan Black B, NADH, MDH and LDH reactions of the testicular interstitium seem correlate to steroid metabolism (biosynthesis and secretion), at the Leydig cells level. The seminiferous epithelium generally had low reactions to all the histochemical studies realized. Particularly in the adbasal compartment the histochemical localizations of NADH diaphorase and LDH were possible related to glycolytic activities and general carbohydrates metabolism, both enzymes, and hydrogen transport, the NADH. The strong PAS, diastase and PAS, and alcian blue pH 2.5 and PAS reactions observed in the adluminal seminiferous epithelium compartment were directly related to the spermatids acrosomal glycoconjugates structuration. Also the SDH localization at this level seems to be related to the mitochondrial activities at the middle piece level in the late spermatids.

Fine structure of the boundary tissue of the seminiferous tubuli of the vampire bat (Desmodus rotundus, G.; Microchiroptera).

Structurally the boundary tissue of the vampire bat seminiferous tubuli showed 2 to 5 layers of connective tissue in which elongated contractile cells and lamellar and/or fibrillar collagen were noticed. This boundary tissue forms the seminiferous tubular lamina propria. Its structure was more complex around the seminiferous tubuli near the Capsula testicularis than between the adjacent and contiguous tubuli into the testicular lobuli. The whole ultrastructural organization of the seminiferous lamina propria was described here.