Focal disruption of spermatogenesis in the testis of adult rats after a single administration of human chorionic gonadotrophin

Summary The effect of a single injection of 100 i.u. human chorionic gonadotrophin (hCG) upon the morphology of the rat testis was studied by light and electron-microscopy from 12–48 h after treatment. Twelve hours after injection of hCG, emigration of leukocytes occurred across the intertubular blood vessels and, both at this time and at 24 h, infiltrations of leukocytes were observed within the extracellular tissue spaces. Furthermore, 12 h after hCG, the seminiferous epithelium showed focal disruption of spermatogenesis involving germ cell degeneration and pyknosis. Focal damage to the seminiferous epithelium persisted at 24 h and 48 h after injection of hCG, the affected seminiferous tubules showing failure of spermiation, accumulation of extracellular vacuoles and degeneration or partial loss of spermatogonia and primary spermatocytes. The observations show that, after stimulation of the Leydig cells with hCG, the intertubular tissue exhibits an inflammatory-type response and this is associated with focal tissue destruction in the seminiferous tubules. It is concluded that a high dose of hCG exerts anti-spermatogenic effects upon the testis and raises the possibility of unexpected interference with tests of normal Leydig cell function in both laboratory and clinical investigations.     

Estradiol and testosterone inhibit rat seminiferous tubule development in a hormone-specific way

Follicle stimulating hormone (FSH), testosterone (T) and estradiol (E₂) are known to regulate testis maturation, and changes in FSH secretion induced by sex steroid treatment may mediate the effects of sex hormones. The aim of this study was to compare the effects of T and E₂ on the pre-meiotic steps of first spermatogenesis and FSH level in rats. Male rat pups were injected daily with 17β-estradiol benzoate (EB; 12.5 μg) or testosterone propionate (TP; 2.5 mg) with the use of one of the two administration modes: 1/transient mode; hormone injections on postnatal days (PND) 1–5 followed by daily vehicle injections until PND 15 (t-EB and t-TP, respectively) or 2/continuous mode; hormone injections on PND 1–15 (c-EB and c-TP, respectively). The control group was injected with vehicle alone. On PND 16, blood was taken for serum hormone measurement and testes were collected for analysis of seminiferous tubule morphometry as well as cell number, proliferation and apoptosis. Testis weight, tubule length, Sertoli and germ cell numbers were reduced, and cell apoptosis in seminiferous epithelium was increased after transient EB and TP treatments. Despite normal or increased FSH secretion, the c-EB treatment inhibited pre-meiotic germ cell development and augmented cell apoptosis, whereas the c-TP treatment reduced the spermatocyte number and inhibited the formation of seminiferous tubule lumen. In conclusion, transient administration of EB or TP during PND 1–5 inhibited testis growth, whereas continuous administration (PND 1–15) impaired pre-meiotic germ cell development in a hormone-specific way.     

Protective effect of exogenous melatonin on testicular histopathology and histomorphometry of adult rats with domperidone-induced hyperprolactinemia

Hyperprolactinemia is a pathological condition resulting from increased prolactin that directly affects reproduction, as this condition inhibits the release of LH, FSH and gonadal steroidogenesis, bringing several negative clinical associations in reproduction. In contrast, melatonin (MEL) plays an important role in the regulation of steroidogenesis and modulates damages to the process of spermatogenesis. The objective was to analyze the protective effects of exogenous melatonin on the testis of hyperprolactinemic adult rats. Forty-eight male rats were used, divided into two treatment periods: 30 and 60 days, each treatment was subdivided into three groups: Control, Hyper (hyperprolactinemia), and Hyper+MEL (hyperprolactinemia and melatonin). Treatment with melatonin was 200 μg/100 g, subcutaneously. Induction of hyperprolactinemia was obtained with a dose of 4 mg/kg of domperidone, subcutaneously. The results of the histopathology demonstrated that the animals in the Hyper group presented degeneration of germ cells when compared to the control. In addition, the degenerations were presented in smaller quantities in the Hyper+MEL, in both treatment periods, evidencing the benefits of the melatonin in gonadal regeneration. The Hyper group of both treatment periods showed a decrease in tubular diameter, epithelium height, and tubular area, in addition to a decrease in Sertoli cells, when compared to the control and the Hyper+MEL group. In conclusion, the hyperprolactinemia can affect the germinal epithelium and testicular microstructure; the exogenous melatonin has a protective effect against hyperprolactinemia, reducing testicular damage.     

In-vitro proliferation of germ cells and supporting cells in the neonatal mouse testis

Summary Testicular cells were prepared from neonatal (48 h after birth) mice by enzymatic dissociation and were cultured in serum-supplemented medium to investigate cell proliferation in vitro. The cultured cells were composed mostly of germ cells, identified by immunocytochemistry using a germ cell-specific antiserum, and supporting (immature Sertoli) cells. After 36 h in culture, the cells were pulse-labeled with ³H-thymidine and fixed at 2-h intervals for 36 h after labeling. Numbers of labeled and unlabeled metaphases of germ cells and supporting cells were counted, and percent labeled metaphases for both cell types were determined for cell-cycle analysis. The results indicate that germ cells, as well as supporting cells, incorporate ³H-thymidine and progress through the cell cycle in vitro. From the curve of the percent labeled metaphases for the supporting cells, the total cell cycle and intervals of DNA synthesis were estimated to be 27.2 h and 13.2 h, respectively.     

Dissociation and identification of intact seminiferous lobules from the testis of the dogfish (Scyliorhinus canicula)

Summary An enzymatic method was developed to obtain intact seminiferous lobules from the testis of the dogfish (Scyliorhinus canicula L.). The freshly isolated lobules were then identified by use of a transillumination technique. Testes from mature dogfish were collected and transverse sections incubated with a mixture of collagenase (0.025%) + pronase (0.08%) at 4° C overnight. Dissociation of the tissue was achieved by mechanical agitation in a calcium and magnesium-free buffer (pH 7.8; 870 mOsm) at room temperature, for 30 min. Based on differences in light absorption and size as well as on comparison with the corresponding histological and ultrastructural cell composition, the seminiferous lobules were identified and classified according to the stages of spermatogenesis of Mellinger (1965). The characteristic changes take place in the size and surface of the lobules and are mainly due to differences in the arrangement of the germ cells, in their number, and in the light absorption characteristics of their nuclei. The combination of the procedures of isolation and transillumination of the dogfish seminiferous lobules, in native condition, offers an original method for the study of germ cell-Sertoli cell interactions in the non-mammalian vertebrate.     

Role of adiponectin as a modulator of testicular function during aging in mice

The mechanisms by which testicular functions decline with aging remain largely speculative. Our recent finding showed the importance of adiponectin in the regulation of testicular functions, whereas its concentration declines during male infertility. Thus, the aim of present study was to explore the potential role of adiponectin during aging. The changes in adiponectin, adiponectin-receptors, and insulin receptor proteins expression in the testis were evaluated and compared with the testicular parameters, mass, and testosterone level in the mice from early post-natal to late senescence period. Further, the current study has examined the effect of exogenous adiponectin treatment on testicular functions in aged mice. The results showed a significant decline in adiponectin/adiponectin-receptors expression simultaneously with a significant decline in testicular mass, insulin receptor expression and testosterone synthesis in the testis of aged mice. Exogenous treatment of adiponectin to aged mice resulted in marked improvements in testicular mass, histological features (cells proliferation), insulin receptor expression, testicular glucose uptake, anti-oxidative enzymes activity and testosterone synthesis as compared with the control. Based on these findings, it may be concluded that a marked decline in adiponectin synthesis and action results in decreased insulin sensitivity (development of insulin resistance) and increased oxidative stress which consequently suppresses testicular functions during aging. This study further showed that treatment with adiponectin ameliorates reduced testicular functions by enhanced expression of insulin receptor in the testis of senescent mice. It is thus hypothesized that systemic adiponectin treatment could be a promising therapeutic strategy for improvement of testosterone production and sperm counts during aging.