Decrease In Diameter Of Seminiferous Tubules Research Articles

Protective effects of CAPE against testicular damage in streptozotocin-induced diabetic rats

Objective: The aim of this study was to investigate the protective effect of CAPE on oxidative stress and apoptosis against streptozotocin (STZ)-induced damage in rat testis after diabetes. Materials and methods: The rats were randomly divided into 4 groups: Control animals, Control animals given CAPE, STZ-induced diabetic animals, and STZ-induced diabetic rats given CAPE. Diabetes was induced by a single intraperitoneal injection of STZ (55 mg/kg). Testicular damage was examined by using hematoxylin and eosin staining and apoptosis was determined by Caspase-3. Potential disorders associated with seminiferous tubular sperm formation were evaluated using the Johnsen score and seminiferous tubule diameters were measured using the Leica Q Win Plus Image Analysis System. Results: Diabetic rats showed an increase in degenerated germ cells along with a decrease in seminiferous tubule diameter. Also, Caspase-3 positive cells were significantly increased in diabetic rats compared to control rats. On the other hand, CAPE significantly reduced the damage and germ cell apoptosis in diabetic rat testis. In testis tissues samples. CAPE treatment significantly decreased the elevated tissue malondialdehyde levels, while increasing the reduced superoxide dismutase enzyme activity. Conclusion: These results suggest that CAPE administered intraperitoneally for 20 days to diabetic rats is a potentially beneficial agent that can be used to reduce testicular damage.

The Effects of Prenatal and Postnatal Exposure to 50-Hz and 3 mT Electromagnetic Field on Rat Testicular Development.

Background and objectives: It has been shown that electromagnetic fields (EMFs) have negative effects on the reproductive system. The biological effects of EMF on the male reproductive system are controversial and vary depending on the frequency and exposure time. Although a limited number of studies have focused on the structural and functional effects of EMF, the effects of prenatal and postnatal EMF exposure on testes are not clear. We aimed to investigate the effects of 50-Hz, 3-mT EMF exposure (5 days/wk, 4 h/day) during pre- and postnatal periods on testis development. Materials and Methods: Pups from three groups of Sprague-Dawley pregnant rats were used: Sham, EMF-28 (EMF-exposure applied during pregnancy and until postnatal day 28), EMF-42 (EMF-exposure applied during pregnancy and until postnatal day 42). The testis tissues and blood samples of male offspring were collected on the postnatal day 42. Results: Morphometric analyses showed a decrease in seminiferous tubule diameter as a result of testicular degeneration in the EMF-42 group. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were decreased in the EMF-42 group. Lipid peroxidation levels were increased in both EMF groups, while antioxidant levels were decreased only in the EMF-28 group. We found decreased levels of vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF1) in the EMF-42 group, and decreased levels of the SRC homology 3 (SH3) and multiple ankyrin repeat domain (SHANK3) in the EMF-28 group in the testis tissue. Conclusions: EMF exposure during pre- and postnatal periods may cause deterioration in the structure and function of testis and decrease in growing factors that would affect testicular functions in male rat pups. In addition to the oxidative stress observed in testis, decreased SHANK3, VEGF, and IGF1 protein levels suggests that these proteins may be mediators in testis affected by EMF exposure. This study shows that EMF exposure during embryonic development and adolescence can cause apoptosis and structural changes in the testis.

EFFECT OF FLAVONOIDS ON THICKNESS AND DIAMETER OF TESTICULAR SEMINIFEROUS TUBULES IN ADULT MALES WISTAR RATES EXPOSED TO OXIDATIVE STRESS BY LEAD ACETATE

The current study aims to determination the effect of flavonoids on thickness and diameter of testicular seminiferous tubules in adult males wistar rats exposed to lead acetate which cause oxidative stress. 40 male wistar rats, each 200±10g on average, were randomly devided to four equal groups and received the following treated for 60 days: First group (C) received nothing except water as a control. Quercetin at dose (300 mg/kg/B.w.) was administered to the second group (T1). Lead acetate at dose (10 mg/kg/B.w.) was administered to the third group (T2). The fourth group received quercetin (300 mg/kg/B.w) for 30 days after receiving lead acetate (10mg/kg/B.w) for 30 days. All of the animals were sacrificed at the end of experiment. Samples of testis, epididymis and vas difference were taken for histopathological study. A significant difference (p≤0.05) in the seminiferous tubule thickness and leydig cell count was reported in a histological examination. sertoli cells, primary and secondary spermatocytes in T1 group in compared with C group. Whereas there were no significant difference in seminiferous tubules diameter in T1 group in compared with the C group. Additionally, there was a significant difference (p≤0.05) between the T2 group and the C group in terms of the thickness of the seminiferous tubules, leydig's cells and sertoli cells number, primary and secondary spermatocytes, and the diameter of the seminiferous tubules. Additionally, there was a significant difference between the T3 group and the T2 group as evidenced by an increase in seminiferous tubule thickness, leydig's cells, sertoli cells, primary and secondary spermatocytes, and a decrease in seminiferous tubule diameter. Also histological sections in control group revealed a complete spermatogenesis, normal testicular tissue architecture with normal seminiferous tubules, epididymis and vas deference tissues. In T1 group there was complete spermatogenesis, the epididymis duct filled with the sperm and extended, vas deference has normal smooth muscles fibers and sperm filled the lumen of it. In T2 group there was vaculation of spermatogonia and few number of sperms, sertoli and leydige cells, the epididymis duct empty with degeneration and destruction of stereocilia, the vas deference empty from sperm and there was mild destruction in the stereocilia and sloughing the epithelial layer of it. In T3 there was large number of spermatogonia, primary and secondary spermatocytes with spermatids, the epididymial duct filled with the sperm .Vas deference has normal smooth muscles and the sperm filled the lumen of it.

Modulatory effects of Punica granatum L juice against 2115 MHz (3G) radiation-induced reproductive toxicity in male Wistar rat.

Advancements in telecommunication sector result in increasing exposure to electromagnetic (EM) radiation, which has been correlated with incidence of male infertility. Therefore, the present study focused on analyzing the consequence of EM radiation (2115 MHz) exposure on the reproductive system of male Wistar rats. Besides, the antioxidant protective effect of Punica granatum juice was also evaluated. For experimental analysis, rats were divided into five groups (control, sham exposed, exposed, herbal plus exposed, and herbal only). Individual group consisted of 6 rats which were exposed to radiation for 45 days (2 h/day). The herbal-treated groups were given 1 ml of Punica granatum extract orally. Various parameters such as organ to body ratio, sperm count, motility, viability, and testis histopathology were studied. Furthermore, oxidative stress parameters and free radical generation were analyzed. The exposed group showed changes in sperm parameters along with decrease in seminiferous tubule diameter. On the contrary, herbal-exposed group showed enhanced sperm count, increased motility, and viability in comparison to exposed group. Histopathology studies also revealed the protective role of herbal juice. Significant alteration in oxidative parameters along with an enhanced free radical generation in exposed group and reduction in herbal groups was observed. The results thus indicate that continuous exposure to EM radiation can lead to oxidative stress which induces biochemical changes in rat sperms. However, Punica granatum extract has a protective role against oxidative damage induced by EM radiation.

Morphological and morphometrical changes on adult Wistar rat testis caused by chronic sodium arsenite exposure.

Arsenic is a contaminant that occurs naturally in the environment, and it is related to several diseases, such as cancer and severe metabolic diseases. Sodium arsenite effects on testes rats are not fully understood regarding morphology and stereology; thus, it becomes necessary to evaluate possible changes in these parameters under low concentrations and simulating occupational exposure. Therefore, the aim of this study was to analyze the morphometrical and stereological changes on rat testis treated with sodium arsenite. The treatment was accomplished using 5mg/kg of sodium arsenite by gastric gavage in Wistar rats, which experiment lasted 8 weeks. Organs were weighed and gonadosomatic index (GSI) was calculated. Using the software Image Pro Plus, seminiferous tubule diameter was measured, and the volume densities of testicular parenchymal components were obtained. It was counted 200 hundred spermatozoa and classified as normal or abnormal. The parameters means of control (N=5) and treated (N=7) groups were compared by U Mann-Whitney's test, and the results were considered significant for P<0.05. We observed a decrease in seminiferous tubule diameter, as well as testis weight. These finds may be related with disorders of testosterone metabolism due to activation of immunological responses of macrophage, which inhibit the steroidogenesis. Thus, we conclude that sodium arsenic does not impair the animal's general health, but its exposure induces biochemical and tissue changes.

Effects of hyper- and hypothyroidism on the development and proliferation of testicular cells in prepubertal rats.

Thyroid hormones are important in the development and regulation of testes. This study was conducted to determine the effects of hyper- and hypothyroidism on testicular development in prepubertal rats aged 20-70days. Weaning male rats (20days old) until day 70 age were randomly divided into four groups: control, hyperthyroid (hyper-T), hypothyroid (hypo-T) and hypothyroid treated with thyroxine (T4) (hypo-T+T4). The results indicated that thyroid hormones caused a significant effect in body and testis weights, and food and water consumption. In addition there were changes in serum concentrations of tri-iodothyronine, T4, thyroid stimulating hormone (TSH) and testosterone. Histomorphology showed a significant decrease in seminiferous tubule diameter in hyper-T compared to the other groups. Leydig cell numbers showed a significant elevation in hyper-T but not in hypo-T groups. Immunostaining indicated that TSH receptor (TSHR), thyroid hormone receptors α/β (TRαβ) and proliferating cell nuclear antigen (PCNA) have the roles in testicular development. Our findings suggest that hyper- and hypo-thyroidism regulate testicular cell proliferation and spermatogenesis in prepubertal rats, indicating that expression of TSHR, TRαβ and PCNA may be regulated by thyroid hormones that are involved in testicular development; and that the administration of T4 to the hypo-T+T4 group leads to an improvement in the testicular condition.

Histopathological Evaluation of Orchiectomy Specimens in 51 Late Postpubertal Men with Unilateral Cryptorchidism

We evaluate the histopathological features of uncorrected undescended testis presenting at a late postpubertal age. The study included 51 men (age 20 to 24 years) diagnosed with inguinal unilateral undescended testis found on routine examination for military recruits. None was evaluated or treated for undescended testis previously. All of the men had a normal contralateral testis and no other observed phenotypic alterations, and all had undergone unilateral orchiectomy. The surgical specimens were first examined histologically, and sections were additionally examined with immunohistochemical methods using antibodies against CD117 and OCT3/4 proteins to verify the presence of intratubular germ cell neoplasia. Histopathology revealed the presence of germ cells at different maturation levels in 26 of 51 (51%) cases. There were 28 cases (55%) with different degrees of basal membrane thickening. A decrease in seminiferous tubule diameter was observed in 23 (45%) patients. Six patients (12%) had dystrophic calcification and 12 (24%) had Leydig cell hyperplasia. Although morphological evaluation did not show intratubular germ cell neoplasia in any patients, 1 with germ cells had positivity for OCT3/4 and CD117 staining. Therefore, 1 case out of51 had diagnosed intratubular germ cell neoplasia. There was a wide range of histopathological changes in undescended testis. Nearly half the patients may still have significant germ cell activity at a variety of maturation levels. The incidence of intratubular germ cell neoplasia was 2% in this group. Intratubular germ cell neoplasia may be overlooked with hematoxylin and eosin staining so immunohistochemical study should be added for evaluation.

2.45-GHz microwave irradiation adversely affects reproductive function in male mouse, Mus musculus by inducing oxidative and nitrosative stress

Electromagnetic radiations are reported to produce long-term and short-term biological effects, which are of great concern to human health due to increasing use of devices emitting EMR especially microwave (MW) radiation in our daily life. In view of the unavoidable use of MW emitting devices (microwaves oven, mobile phones, Wi-Fi, etc.) and their harmful effects on biological system, it was thought worthwhile to investigate the long-term effects of low-level MW irradiation on the reproductive function of male Swiss strain mice and its mechanism of action. Twelve-week-old mice were exposed to non-thermal low-level 2.45-GHz MW radiation (CW for 2 h/day for 30 days, power density = 0.029812 mW/cm2 and SAR = 0.018 W/Kg). Sperm count and sperm viability test were done as well as vital organs were processed to study different stress parameters. Plasma was used for testosterone and testis for 3β HSD assay. Immunohistochemistry of 3β HSD and nitric oxide synthase (i-NOS) was also performed in testis. We observed that MW irradiation induced a significant decrease in sperm count and sperm viability along with the decrease in seminiferous tubule diameter and degeneration of seminiferous tubules. Reduction in testicular 3β HSD activity and plasma testosterone levels was also noted in the exposed group of mice. Increased expression of testicular i-NOS was observed in the MW-irradiated group of mice. Further, these adverse reproductive effects suggest that chronic exposure to nonionizing MW radiation may lead to infertility via free radical species-mediated pathway.

Melatonin and Testicular Damage in Busulfan Treated Mice

Background:Advancement in the treatment of various types of cancer has led to greater patient survival. These treatments essentially have toxic effects on different kinds of cells, such as germ cells. Infertility as one of the side effects of cancer treatment has changed the quality of life of young cancer survivors dramatically. Melatonin is an antioxidant with receptors in the reproductive systems.Objectives:We supposed that melatonin, as an antioxidant, may protect testis against the toxic effects of the drugs.Materials and Methods:In this experimental study, three groups with seven mice each, were allocated. The control group received normal saline for two months, and the busulfan group received a single dose of 40 mg/kg busulfan intra-peritoneally, and the melatonin group received 20 mg/kg melatonin daily for two months, 45 days after a single dose of busulfan. Next, after decapitation and removal of the testis, tissues were fixed in Bouin's solution and stained by H&E and TUNEL. The sections were evaluated, assessing morphology and spermatogenesis.Results:In this research, a significant reduction in Johnson’s criteria in the busulfan group (Mean rank = 15.50) was found versus the control group (Mean rank = 45.50), P < 0.001 and in the melatonin group (Mean rank = 45.50) compared to the busulfan group (Mean rank = 15.50), P < 0.001. There was a significant difference between the melatonin and control groups, P < 0.05. In addition, a significant decrease in seminiferous tubule diameter was observed in the busulfan group (763.2 ± 104.41) versus the control group (855.4 ± 52.35), P < 0.01 and melatonin group (834.2 ± 87.26), P < 0.05. Testicular epithelium height was significantly decreased in the busulfan group (Mean rank = 14.60) compared to the control group (Mean rank = 26.40), P < 0.01 and in the busulfan group (Mean rank = 14.95) in comparison with the melatonin group (Mean rank = 26.05), P < 0.01. Also melatonin group (Mean rank = 25.42) showed a significant reduction in epithelium height compared to the control group (Mean rank = 35.58), P < 0.05. Spermatogenesis was impaired in the busulfan group. Although melatonin reduced the rate of apoptosis in the busulfan group, yet it could not remove all apoptotic cells.Conclusions:This study indicated that melatonin ameliorates the cytotoxic effects of busulfan on germ cells.

The effect of zinc oxide nanoparticles on mouse spermatogenesis

To evaluate the effects of zinc oxide nanoparticles on mouse spermatogenesis. Thirty two adult male NMRI mice were used. Experimental Groups (ZNP-1-ZNP-3) received one of the following treatments daily for 35days: 5, 50 and 300mg/kg zinc oxide nanoparticles respectively. Control group received only distilled water. Epididymal sperm parameters, testicular histopathology, morphometric analysis and spermatogenesis assessments were performed for evaluation of the zinc oxide nanoparticles effects on testis. Epididymal sperm parameters including sperm number, motility and percentage of abnormality were significantly changed in 50 and 300mg/kg zinc oxide nanoparticles treated mice (p < 0.01). Histopathological criteria such as epithelial vacuolization, sloughing of germ and detachment were significantly increased in 50 and 300mg/kg zinc oxide nanoparticles treated mice (p < 0.001). 300mg/kg zinc oxide nanoparticles induced formation of multinucleated giant cells in the germinal epithelium. 50 and 300mg/kg zinc oxide nanoparticles also caused a significant decrease in seminiferous tubule diameter, seminiferous epithelium height and maturation arrest (p < 0.001). Zinc oxide nanoparticles act as testicular toxicant and further studies are needed to establish its mechanism of action upon spermatogenesis.

Impact of moderate calorie restriction on testicular morphology and endocrine function in adult rhesus macaques (Macaca mulatta)

We previously reported that moderate calorie restriction (CR) has minimal impact on testicular gene expression in young adult rhesus macaques, and no obvious negative impact on semen quality or plasma testosterone levels. We now extend these findings by examining the influence of CR on various aspects of the reproductive axis of older males, including 24-h circulating testosterone levels, testicular gene expression, and testicular morphology. Young adult and old adult male rhesus macaques were subjected to either 30% CR for 5-7years, or were fed a standard control diet. Analysis of the 24-h plasma testosterone profiles revealed a significant age-associated decline, but no evidence for CR-induced suppression in either the young or old males. Similarly, expression profiling of key genes associated with testosterone biosynthesis and Leydig cell maintenance showed no significant CR-induced changes in either the young or old animals. The only evidence for CR-associated negative effects on the testis was detected in the old animals at the histological level; when old CR animals were compared with their age-matched controls, there was a modest decrease in seminiferous tubule diameter and epithelium height, with a concomitant increase in the number of depleted germ cell lines. Reassuringly, data from this study and our previous study suggest that moderate CR does not negatively impact 24-h plasma testosterone profiles or testicular gene expression. Although there appear to be some minor CR-induced effects on testicular morphology in old animals, it is unclear if these would significantly compromise fertility.

Toxic effects of Carthamus tinctorius L. (Safflower) extract on mouse spermatogenesis.

To evaluate the effects of aqueous extract of Carthamus tinctorius L., also named safflower, on mouse spermatogenesis. Sixteen adult male NMRI mice were used. Experimental group received Carthamus tinctorius L. extract at the dose of 200 mg/kg for 35 consecutive days and control group received only distilled water. Testicular histopathology, morphometric analysis and spermatogenesis assessments were performed for evaluation of the Carthamus tinctorius L. extract effects on testis. Histopathological criteria such as epithelial vacuolization, sloughing of germ and detachment were significantly decreased in Carthamus tinctorius L. treated mice (p < 0.001). Carthamus tinctorius L. extract induced formation of multinucleated giant cells in the germinal epithelium. Carthamus tinctorius L. extract also caused a significant decrease in seminiferous tubule diameter, seminiferous epithelium height and maturation arrest (p < 0.001). Carthamus tinctorius L. extract has toxic effects on mouse testicular tissue, and recommended to use it with caution if there is a reproductive problem.

Seasonal Changes in Spermatogenesis (Including Germ Cell Degeneration) and Plasma Testosterone Concentration in the Grey-headed Fruit Bat, Pteropus poliocephalus

Maximal testicular size of P. poliocephalus occurred during the February-March-April period (autumn breeding season) associated with maximal plasma levels of testosterone (121' 5 � 23� 4 nmol/I). Testicular size decreased after the breeding season concomitant with a decrease in plasma testosterone (7'6 � 1�7 nmol/I). Histologically, regressive changes were observed in the testis after the breeding season including a decrease in seminiferous tubule diameter and a decrease in Leydig cell nuclear diameter. Significant germ cell loss occurred during the breeding season (44%) mainly during the final spermatogonial division and meiosis and increased in regressed testes (69%), being accentuated chiefly at meiosis. All regressed testes showed some sperm production although it was much reduced after the breeding season; however, optimal fertility appears to be restricted to autumn by the large increases in testosterone secretion which only occur at this time.