Karine Moura de Freitas

Histomorphometric evaluation of the neotropical brown brocket deer Mazama gouazoubira testis, with an emphasis on cell population indexes of spermatogenic yield

Information on the reproductive biology of neotropical cervids is scarce. Therefore, the aim of this study was to perform biometric, histologic and stereologic analyses of the brown brocket deer Mazama gouazoubira testis, with an emphasis on the intrinsic yield and the Sertoli cell index. Seven adult males kept in captivity were used. The animals were immobilized; anesthetized and testicle fragments were obtained by biopsy incision. The material was fixed, processed and examined by routine histological methods for light microscopy. The average body weight was 17.2kg, from which 0.40% were allocated in gonads and 0.33% in seminiferous tubules, which represented 85.9% of the testis parenchyma. The mean albuginea width and volume were 345.7μm and 3.5mL (5.3% of the testicular weight), respectively. The mean mediastinum volume of both testicles was 1.0mL (1.5% of the testicular weight) and the testicular parenchyma volume corresponded to 93.1% of total testicular weight (64.9g). The seminiferous tubules diameter was 224.4μm, while the epithelium height was 69.6μm. On average, an adult brown brocket deer showed a total of 1418m of seminiferous tubules in both testicles (21.5m per gram of testis). Each stage I seminiferous tubular cross section contained 1.10 type A spermatogonia, 13.4 primary spermatocytes in pre-leptotene/leptotene, 13.7 spermatocytes in pachytene, 48.8 round spermatids and 3.7 Sertoli cells. The general yield of spermatogenesis was 44.7 cells and the Sertoli cell index was 13.2. The qualitative and quantitative description of testicular histology of brown brocket deer help to understand its spermatogenic process and to establish parameters for the reproductive biology of this wild species. Furthermore, the data from the present research will help further studies using other species of Brazilian cervids, especially endangered ones, making an additional effort to the species preservation.

Association of the infusion of Heteropterys aphrodisiaca and endurance training brings spermatogenetic advantages

The species Heteropterys aphrodisiaca is commonly used as a stimulant by popular medicine in the Cerrado, a savanna-like biome, Brazil. Recent studies have proved its protective effects on testes of animals submitted to treatment using Cyclosporine A, as well as its stimulus effect in increasing testosterone secretion. Therefore, the present study was designed to analyze whether the association of the plant infusion and endurance exercise could potentiate the stimulating effect. The animals were separated into 4 groups: two control (sedentary and trained) receiving water and two treated (sedentary and trained) receiving the plant infusion daily (104 mg/day). The proportion of the seminiferous tubule compartment and interstitium was analyzed. Within the seminiferous epithelium, the number of Sertoli and germ cells were counted in order to evaluate whether the treatment would alter the spermatogenic dynamics, analyzing: the spermatogenic yield, the mitotic and meiotic indexes, the total number of germ cells and the Sertoli cell support capacity. Trained and treated animals showed increased spermatogenic yield and spermatogonia mitosis, and no significant differences in apoptotic indexes. Despite the results showing the same pattern regarding yield and mitotic index, the meiotic index was higher in the sedentary/treated group. Therefore, the H. aphrodisiaca infusion increased both the testosterone production and the spermatogonia mitosis, thus increasing the spermatogenic yield.

Histomorphometric evaluation of the Molossus molossus (Chiroptera, Molossidae) testis: The tubular compartment and indices of sperm production

Insectivorous bats play a very important role in the regulation of tropical ecosystems, but information about their reproductive cycle is lacking. Thus, male Molossus molossus were captured over the four seasons, and morphometric analyses of their testes were conducted to infer on the gonadal dynamics and the reproductive capacity of the species. Testes were immersed in Karnovsky fixative, and fragments were embedded in methacrylate and paraplast for morphometric and TUNEL assay respectively. The least gonadosomatic index (0.3%), tubulesomatic index (0.2%) and tubular diameter (133.2μm) occurred in summer. An adult M. molossus showed a total average of 48.9m of seminiferous tubules per gram of testis. Primary spermatocytes were observed in the zygotene at Stage 1 of the seminiferous epithelium cycle. The greatest meiotic index was obtained in winter (3.8 cells), and the general yield of spermatogenesis was higher in winter (64.5 cells) than in summer (19.1 cells). There was no difference in the apoptotic cells count among seasons. The Sertoli cell index was less in summer (5.9) than in fall (11.6), while the number of Sertoli cells per gram of testis did not vary significantly among the seasons (28.0×10(7)). The spermatic reserve per gram of testis was greater in the fall (63.9×10(7)) and winter (69.8×10(7)) than summer (37.1×10(7)). We conclude that M. molossus males show a continuous reproductive cycle, featuring greater spermatogenic activity during the fall and winter, a tubular length above the average of other mammals and a less support capacity of the Sertoli cells.

Acupuncture increases the diameter and reorganisation of collagen fibrils during rat tendon healing

Background Our previous study showed that electroacupuncture (EA) increases the concentration and reorganisation of collagen in a rat model of tendon healing. However, the ultrastructure of collagen fibrils after acupuncture is unknown. Objectives To assess the effect of acupuncture protocols on the ultrastructure of collagen fibrils during tendon healing. Methods Sixty-four rats were divided into the following groups: non-tenotomised (normal group), tenotomised (teno group), tenotomised and subjected to manual acupuncture at ST36 (ST36 group), BL57 (BL57 group) and ST36+BL57 (SB group) and EA at ST36+BL57 (EA group). The mass-average diameter (MAD) and the reorganisation of collagen fibril diameters were determined during the three phases of tendon healing (at 7, 14 and 21 days). Results The MAD increased during the three phases of healing in the SB group. In the EA group, MAD increased initially but was reduced at day 21. The reorganisation of collagen fibrils was improved in the EA and SB groups at days 14 and 21, respectively. EA at day 21 appeared to reduce the reorganisation. Conclusions These results indicate that the use of EA up to day 14 and manual acupuncture at ST36+BL57 up to day 21 improve the ultrastructure of collagen fibrils, indicating strengthening of the tendon structure. These data suggest a potential role for acupuncture in rehabilitation protocols.

Organization and seasonal quantification of the intertubular compartment in the bat Molossus molossus (Pallas, 1776) testis

Environmental factors can influence the reproductive rates in bats, and since morphometric information of bats testis is scarce, we aimed to compare the organization and quantification of the intertubular components in the testes of the bat Molossus molossus, collected in different seasons. Testicular histological sections were evaluated using light and electron microscopy. The intertubular compartment occupied an average 10% of the testes, being predominately constituted of Leydig cells (LC). The percentages of the testes occupied by the intertubular compartment and by LC were significantly higher in summer, while the other intertubular components did not vary significantly among the seasons. As suspected under light microscopy, the ultrastructural analysis confirmed the existence of multinucleated LC during winter. The increase in the nuclear percentage of LC in winter seems to have caused the decrease of the cytoplasmatic measurements in that season, as well as in the volume of LC. The highest cytoplasmatic values and volume of LC registered in the spring, summer, and fall can be related to greater activity of this cell in these seasons. The higher investment in intertubular tissue and in LC observed in summer, compared to winter; suggest an increase in the steroidogenic capacity of this bat during summer. The analyses correlating testicular morphometry and abiotic environmental factors in this study confirm the influence of climatic factors on the reproduction of M. molossus. Microsc. Res. Tech., 2013.

Cycle of the seminiferous epithelium of the bat Molossus molossus, characterized by tubular morphology and acrosomal development

Abstract Objective To describe the seminiferous epithelium cycle (SEC) by tubular morphology method, and the acrosomal development of individualizing spermatids, and to explore the distinction of the stages between two generations of spermatids. Methods Testicular fragments were fixed in Karnovsky, embedded in glycol methacrylate and examined under light microscopy and transmission electron microscopy. Sections in 3 μm thickness were stained with toluidine blue for the characterization of the stages of the SEC by the tubular morphology method, or submitted to the PAS reaction for the visualization of the acrosomal formation. Additional details on the acrosomal formation were observed under transmission electron microscopy. Results Through the eight stages described by tubular morphology method, 10 steps of acrosomal formation were observed in the spermatid development, called acrosomal steps. As the spermatids were produced in stage V of the tubular morphology method, it was at this stage from which began the steps of acrosomal development. Conclusions We propose association of the acrosomal steps for the first time, with the different stages by tubular morphology method. This method presents an alternative to the existent Methods, allowing interspecific comparisons of the SEC, not only among different species of bats, but also among the other mammals.

The effects of Cyclosporin A and Heteropterys tomentosa on the rat liver

Cyclosporin A (CsA) is a widely employed immunosuppressive drug that is associated with several side effects, among then hepatotoxicity. Heteropterys tomentosa is a Brazilian plant efficient in reducing damage caused by CsA on the rat testis and prostate. The aim of this study was to evaluate the effect of CsA and H. tomentosa (administered isolated or simultaneously) on the liver of Wistar rats. The animals were treated daily with water (control), CsA (15 mg/kg/day), H. tomentosa infusion or CsA+H. tomentosa, for 21 or 56 days. The treatments did not alter liver morphology or cause fibrosis. H. tomentosa administered for 21 days increased the number of hepatocyte nuclei and Kupffer cell volumetric proportion. After 56 days of treatment, H. tomentosa administration did not alter the parameters analyzed. Biochemical plasma dosages and liver stereology showed impairment caused by CsA-treatment after 21 days; these results were not observed after 56 days of treatment. The simultaneous treatment with CsA and H. tomentosa for 21 or 56 days did not alleviate nor accentuate CsA hepatic effects. The present study showed that the 21 days treatment with CsA caused more alteration to the liver than the 56 days treatment; this could be related to hepatic recovery after the long term treatment.

Advantage of Guarana´ (Paullinia cupana Mart.) Supplementation on Cadmium-induced Damages in Testis of Adult Wistar Rats

Paullinia cupana is an Amazonian bush whose seeds have long been used in folk medicine. However, most of the therapeutic properties attributed to this plant are broad and nonspecific, although an antioxidant activity has been reported. On the other hand, cadmium is a heavy metal known for increasing free radicals, hence resulting in cellular oxidative damages. This study was designed to evaluate whether Paullinia cupana is able to reduce cadmium-induced morphological impairment in Wistar rat testis. Adult male Wistar rats 110 days old were ip injected with cadmium (1.15 mg/kg BW [body weight]) and subsequently treated with P. cupana during 56 days. Furthermore, groups receiving either P. cupana extract or cadmium are mentioned. After the treatment period, testis samples were subjected to histological and stereological analyses. Moderate to severe testicular impairments were shown by the animals exposed to cadmium. However, the animals supplemented with P. cupana after cadmium exposure showed a significant decrease in the proportion of damaged seminiferous tubules. Also, P. cupana supplementation was effective in maintaining the number of Leydig cells per testis in the animals exposed to cadmium. In conclusion, P. cupana supplementation was partially efficient in preventing cadmium from damaging the testis of adult Wistar rats.

Effects of Catuaba Cristal ® on the Testis of Wistar rats

The aim of this study was to evaluate the possible beneficial effects of Catuaba Cristal® (CC), an alcoholic drink made from wine and Erythroxylum catuaba Ar. Cam on testis. Wistar rats either received CC solution (n=8) or water (n=9). Results showed significant body weight reduction within the CC group, although, no weight changes were observed for liver, kidney, testis, epididymis, seminal vesicle and prostate. The volumetric proportion and volume of interstitial tissue and lymphatic space were reduced in the treated group. In the CC group, although the nuclear volume of Leydig cells (LC) decreased, the number of LC per testis increased. These results suggested that CC had no beneficial effect on spermatogenesis of Wistar rats.

Heteropterys tomentosa (A. Juss.) infusion counteracts Cyclosporin a side effects on the ventral prostate

BackgroundCyclosporin A (CsA) is an immunosuppressive drug widely used in treatment of auto-immune diseases or after organ transplants. However, several side effects are commonly associated with CsA long term intake, some regarding to loss of reproductive organ function due to oxidative damage. Considering that phytotherapy is an important tool often used against oxidative stress, we would like to describe the beneficial effects of Heteropterys tomentosa intake to minimize the damage caused by CsA to the ventral prostate tissue of Wistar rats under laboratorial conditions.MethodsThirty adult Wistar rats (Rattus norvegicus albinus) were divided into: control group (water); CsA group (Cyclosporin A); Ht group (H. tomentosa infusion) and CsA + Ht group (CsA and H. tomentosa infusion). Plasmic levels of hepatotoxicity markers, triglycerides, cholesterol and glucose were quantified. The ventral prostate tissue was analyzed under light microscopy, using stereological, morphometrical and immunohistochemical techniques.ResultsH. tomentosa did not cause any alterations either of the plasmic parameters or of the ventral prostate structure. CsA caused alterations of GOT, total and indirect bilirubin, cholesterol, triglycerides and glucose levels in the plasma; CsA-treated rats showed alterations of the ventral prostate tissue. There were no alterations regarding the plasma levels of GOT, triglycerides and glucose of CsA + Ht animals. The same group also showed normalization of most of the parameters analyzed on the ventral prostate tissue when compared to the CsA group. The treatments did not alter the pattern of AR expression or the apoptotic index of the ventral prostate epithelium.ConclusionsThe results suggest a protective action of the H. tomentosa infusion against the side effects of CsA on the ventral prostate tissue, which could also be observed with plasmic biochemical parameters.