Basal Lamina Of Seminiferous Tubules Research Articles

COenzyme Q10 ameliorates bisphinol A induced reproductive male toxicity

Background and aim The endocrine disruptor compound bisphenol A (BPA) affects spermatogenesis and exacerbates benign prostate hyperplasia induced by testosterone. Nonetheless, the direct effect of BPA on prostate and testicular cells is not fully investigated. The objective of this study was to evaluate the pathogenic effects of BPA on the structure of the prostate and the ultrastructure of the testis of adult male albino rats via immunohistochemical and transmission electron microscopic study and the potential protective effect of CoQ10 supplementation. Methods A total of 30 male albino Wistar rats were categorized into five equal cohorts: group I, no treatment; group II was administered corn oil; group III received coenzyme Q10 (CoQ10); group IV was administrated BPA; and group V received BPA+CoQ10. Results BPA administration significantly decreased the mean values of the plasma fertility hormones and serum antioxidant enzymes and increased malondialdehyde. BPA administration markedly affected seminal parameters. Coadministered CoQ10 significantly reversed these biochemical changes. BPA induced histopathological alterations in the epithelium and connective tissue of prostate. Immunohistochemistry of the prostate revealed decreased E-cadherin and increased vimentin expressions in BPA-treated group. Ultrastructural analysis of the testis showed impairment of the basal lamina of seminiferous tubules and tight junctions between Sertoli cells after BPA exposure. Conclusion The biochemical and histopathological results of this study revealed direct evidence for BPA-induced male reproductive toxicity in the testes and prostate, causing male infertility. CoQ10 coadministration with BPA partially protects against its damaging effect mediated via its antioxidant capabilities.

Influence of hormonal imbalance on the integrity of seminiferous epithelium in the testes of adult rats chronically exposed to letrozole and rats exposed to soya isoflavones during the prenatal period, lactation, and up to sexual maturity

The structural integrity of the germ cells in the seminiferous epithelium and the correct process of spermatogenesis are made possible by proteins that participate in the formation of different types of junctions. This study was performed on samples of the testes of 4 groups (2 experimental and 2 corresponding control) of male Wistar rats. In the first experimental group, the adult rats received letrozole – a nonsteroidal inhibitor of cytochrome P450 aromatase (P450arom). The second experimental group was exposed to soya isoflavones during the prenatal period, lactation, and up to sexual maturity. The aim of this study was to examine the immunoexpression of β-catenin, N-cadherin, occludin, connexin43, annexin V, and advanced glycation end products (AGE) in the seminiferous epithelium of rat testes with chronic estrogen deficiency and of rats exposed to soya isoflavones. Series of sections of the testes were stained using PAS and silver impregnation. Moreover, immunohistochemistry tests were performed. A semi-quantitative determination of protein immunoexpression was performed using Image J. The number of annexin V positive Sertoli cells per tubule were counted manually. Comparisons between the experimental and corresponding control groups were performed using a non-parametric Mann-Whitney U test. The most common alterations were prematurely sloughed germ cells in the lumen of the seminiferous tubules and invaginations of the seminiferous tubules. We observed a lower number of annexin V positive Sertoli cells and a lower expression of N-cadherin and occludin in the seminiferous epithelium of both groups of rats with hormonal imbalances. Moreover, a higher expression of AGE, a lower expression of connexin 43 and a lower amount of reticular fibers in the basal lamina of seminiferous tubules was present in rats treated with letrozole and a higher expression of β-catenin was found in rats exposed to soya isoflavones. The hormonal imbalance between androgens and estrogens resulted in a decreased number of annexin V positive Sertoli cells. This may be associated with a failed clearance of apoptotic germ cells that leads to disturbances in the blood-testis-barrier (BTB) by affecting the expression of junctional proteins in the seminiferous epithelium. Moreover, a decreased level of estrogens was also associated with an increased expression of AGEs and with a changed composition of basal lamina in the seminiferous tubules of rats. These changes could lead to germ cell sloughing and invaginations of the seminiferous tubules.

Consequences of Microwave oven Exposed Diet on The Basal Lamina of Seminiferous Tubules of Mice

Usage of electronic gadgets like microwave oven is increasing day by day that heats the food by exposing it to electromagnetic radiations which has many hazardous effects on human health including fertility. Aim: To find the effects of microwave oven exposed diet on basal lamina of seminiferous tubules of mice alongwith protective effects of Mentha piperita and melatonin on the same tissue. Study Design: Randomized control trial. Methodology: Adult male mice (n=32) were divided into four groups. Control group (G1) received standard pellets prepared for mice. Second group (G2) was given mice pellets exposed to microwave oven. Third group (G3) received Mentha Piperita leaf extract along with mice pellets exposed to microwave oven and the fourth group (G4) received oral melatonin along with pellets exposed to microwave oven. Later their testicular tissue was removed for histological examination while basal lamina disruption was assessed by scoring. Data analyzed by SPSS 22.0v. Results: In group G2, there was slight disruption in the basal lamina in 75% of the cases while in experimental group G3, there was slight disruption of basal lamina only in 12.5% of the cases. However, in group G4, only 25% specimen had slight disruption of basal lamina Conclusion: It was concluded that microwave oven exposed diet produced severe disruption of basal lamina in group G2 that decreased in Mentha piperita and melatonin treated groups. However, Mentha piperita treated group produced better results than melatonin treated group. Keywords: Mice, Testis, Basal Lamina, Mentha piperita and Melatonin

Localization of Laminin and Fibronectin in Rat Testes after Diisobutyl Phthalate Exposure: Histopathologic and Immunohistochemical Study

In this study the effects of Diisobutyl phthalate (DIBP) from commonly used pytalates were histopathologically investigated in order to find out whether it is one of the causes of increased infertility in recent years. In addition, the localization of laminin and fibronectin was demonstrated immunohistochemically after exposure to DIBP. For this purpose, three study groups from Wistar albino rats were created: experimental group, control group and corn oil applied group. The experimental group were administered by gavage 3 different doses (0.25-0.5-1 ml/kg/day) of DIBP mixed with corn oil every day for 28 days. At the end of the experiment, the testis tissue samples from all of the experimental and control group animals were investigated and evaluated with a light microscope after routine histological and immunohistochemical preparation processes. Reduction in spermatogenic cell population, apoptotic and necrotic germ cells, increase in Sertoli cells, multinucleate giant cells, thickening and dilatation in basal lamina of seminiferous tubules, oedema and shrinkage in interstitial connective tissue were observed in DIBP treatment groups. The decrease in localization of fibronectin and laminin immunoreactivity in seminiferous tubules was observed. The results of this study demonstrated that DIBP causes serious damage to the testicular tissue, especially in terms of spermatogenesis. For this reason, it has come to the conclusion that the material in question may cause infertility in male individuals.

The applications and recovery outcome of spermatogonia stem cells in regenerative medicine

Abstract Spermatogonial stem cells are unique cells that line the basal lamina of seminiferous tubules in the testis and have the potential of self-renewal and differentiation. These cells pass on genetic information to the next generation via the process of spermatogenesis and have an important role in the maintenance of male fertility. Recent reports have shown the pluripotency characteristics of spermatogonial stem cells at the cellular and molecular level. These cells can convert into Embryonic stem like cells that exhibit similar phenotype with embryonic stem cells and the capacity to differentiate into all three germ layers: Ectoderm, endoderm and mesoderm. These properties make spermatogonial stem cells an appropriate source for use in regenerative medicine. Spermatogonial stem cells considered as an alternative and novel cell source that limits the technical and ethical problems accompanied with the other pluripotent stem cells. Patient-specific cell lineages without limitations of immunological rejection and specific auto transplantation are the beneficial’s associated with these cell population. Also, cell therapy based on spermatogonial stem cells engraftment, promises for preservation of fertility and induces the reproductive potential in infertile patients. Here, we discuss the applications of spermatogonial stem cells in regenerative medicine with focus on all important aspects, including: cell isolation, propagation, differentiation and especially cell transplantation.

Histologic study of testis injury after bisphenol A exposure in mice.

The health effects of bisphenol A (BPA) have become a great concern in recent years. In this study, the reproductive toxicity of BPA was investigated. Male CD-1 mice were orally administrated with BPA (0, 100, 300 and 600 mg kg-1 body weight) for 56 consecutive days. Results showed that relative testis weight to total body weight was significantly lower in the high-dose group ( p < 0.01, p < 0.05). Microscopic examination under light and transmission electron microscopes showed disorders of spermatogenesis after BPA exposure, including rough basal lamina of seminiferous tubules and damage of tight junctions between Sertoli cells. Further study by terminal-deoxynucleoitidyl transferase-mediated nick end labelling assay showed a significant induction of apoptosis in the testis tissue of the BPA groups ( p < 0.01). Immunohistochemical study found that the expression of androgen-binding protein (ABP) was significantly decreased in BPA-treated mice ( p < 0.01). Our results indicated that impairment of the basal lamina of seminiferous tubules and tight junctions may contribute to BPA-induced cell injury. A decrease in the level of ABP could be the possible mechanism for the reproductive toxicity of BPA. These findings provided direct evidence and novel insight into the reproductive toxicity of BPA and may have implications for understanding the toxicity of other endocrine disruptors.

Effects of 50 Hz Magnetic Field on the Testes and Sperms of Adult Albino Rats

This study evaluates the 'effect of 50 Hz magnetic field' on the testicular gross morphology and histology, sperm morphology and meiotic chromosomes of adult Albino rats. The rats were divided into three groups; Control and two exposed groups to 0.15 mT and 20 mT. Thirty seven rats were included in each group. The exposure time lasted for 2 weeks, 3 days/week and 1/2 hour/day. After exposure, the rats were sacrificed by cervical dislocation and handled for evaluation. One of the testes was examined for gross morphological and histopathological changes. The other one was handled for evaluation of meiotic chromosomal abnormalities in the spermatocytes. The sperms were extracted from the epididymis and vas deferens for morphological investigation. There were no observed gross morphological changes. The histopathological evaluation revealed a picture of maturation arrest at the level of the round spermatids with slight degenerative changes. These changes were more marked at the third group. The morphometric study showed reduction in the cellular surface area of the seminiferous tubules and an increase in the thickness of the basal lamina of seminiferous tubules, the 'number of the Leydig cells in the interstitial space' and the surface area of blood vessels. There was no effect on either the seminiferous tubular diameter, nucleo-cytoplasmic ratio of Leydig cells or the Sertoli cell number. The sperm morphological evaluation showed increase in the number of abnormal forms due to exposure. The chromosomal investigation showed also an increase in the chromosomal abnormalities after exposure.

Mice Stage-Specific Claudin 3 Expression Regulates Progression of Meiosis in Early Stage Spermatocytes1

Claudin 3 is a protein component of the tight junction strands. Tight junctions between adjacent Sertoli cells form the blood-testis barrier (BTB). During spermatogenesis, seminiferous stage-specific expression of claudin 3 is believed to regulate the migration of preleptotene/leptotene spermatocytes across the BTB. Here, we determined the cell types expressing claudin 3 in adult mouse testis and investigated spermatogenesis after testis-specific in vivo knockdown of claudin 3. The results of in situ hybridization revealed that claudin 3 mRNA was predominantly expressed in germ cells near the basal lamina of seminiferous tubules at stages VI-IX. Furthermore, claudin 3 protein was localized not only to the BTB but also to the cell membrane of STRA8-expressing preleptotene/leptotene spermatocytes in the testis of adult ICR.Cg-Tg(Stra8-EGFP)1Ysa/YsaRbrc mice. Although claudin 3 knockdown did not affect BTB integrity, it did cause a partial delay in spermatocyte migration across the BTB. Moreover, claudin 3 knockdown resulted in a prolonged preleptotene phase during spermatogenesis. These data indicate that the seminiferous stage-specific expression and localization of claudin 3 during spermatogenesis regulate the progression of meiosis by promoting germ cell migration across the BTB.

The Niche for Spermatogonial Stem Cells in the Mammalian Testis

The theory of the "stem cell niche" was originally proposed for the hematopoietic system, and the existence of the niche as an actual entity was proved in the Drosophila germ cell system. Historically, mammalian spermatogenesis has been studied extensively as a prime example of a stem cell system, and studies have established a stem-progenitor hierarchical order of spermatogonia. In the niche on the basal lamina of seminiferous tubules, spermatogonial stem cells (SSCs) are secluded from the outside world and divide constantly to self-renew and differentiate. During the last 10 years, the development and exploitation of the germ cell transplantation method has expanded our understanding of the nature of SSCs and their niches. The ability to maintain and expand SSCs in vitro, which recently became possible, has further reinforced this research area as a mecca of stem cell biology. Nonetheless, the mammalian germ stem cell and its niche remain to be defined more strictly and precisely. We are still on a journey in search of the real stem cell and its true niche.

Leydig cells within the aspermatogenic seminiferous tubules

Cells identical to Leydig cells were found within a peritubular boundary layer and even inside a basal lamina of seminiferous tubules in three male patients (two with inguinal cryptorchism and one with infertility). The seminiferous tubules of all patients showed a moderate to marked thickening of the boundary layer and a complete loss of spermatogenic cells. The "ectopic Leydig cells" were characterized by the presence of Reinke crystals or an extensively developed smooth endoplasmic reticulum. These cells were believed to have differentiated in situ from myoid cells within the boundary layer and also to have invaded from the interstitial tissue in the form of mature Leydig cells. The occurrence of ectopic Leydig cells appeared to parallel the extent of loss of the Sertoli cells and also that of the thickening of the boundary layer. The functional significance of the ectopic occurrence might be implicated in the impaired spermatogenesis.

Spermatogenic disturbance induced in mice by combined local injection of monoclonal antibodies to Sertoli cell and to basal lamina of seminiferous tubule.

Two kinds of hybridoma clones, one producing monoclonal antibodies against Sertoli cell (TM-1) and the other the basal lamina of the seminiferous tubule (TM-2), were raised by fusion between P3X63Ag8-653 mouse myeloma cells and spleen cells of BALB/c mice (H-2d) immunized with testicular homogenate of the same inbred mice. Immunohistochemically, TM-1 reacted specifically with cytoplasmic component of Sertoli cell and TM-2 with basal lamina of the seminiferous tubule. Using these monoclonal antibodies, spermatogenic disturbance was induced experimentally in BDF1 (H-2b/d) by intratesticular injection of a set of these two antibodies. Single injection of either TM-1 or TM-2 failed to induce the lesion. This fact indicated that TM-1 antibody could reach the Sertoli cell to impair its function, which was otherwise inaccessible without coincidental action of TM-2 antibody. TM-2 antibody appeared to alter the permeability of the basal lamina of the tubule and lower its barrier effect.

Pathogenesis of Trypanosoma brucei infection in deer mice (Peromyscus maniculatus). Light and electron microscopic study of testicular lesions.

The results of light and electron microscopic studies demonstrated that Trypanosoma brucei infection produced marked testicular degeneration in deer mice (Peromyscus maniculatus) with highly significant (p less than 0.001) decreases in testicular weight, seminiferous tubular diameter, and epithelial thickness. Lesions in seminiferous tubules consisted of necrosis of spermatids, spermatocytes, and sometimes the spermatogonia. Sertoli's cells had marked vacuolation and accumulation of phagocytic material. Orchitis was present in most infected mice, particularly those with severe seminiferous tubular degeneration. There was marked folding of the basal laminae of the seminiferous tubule and myoid layer, and a consistent increase in the layers of the basal laminae of the seminiferous tubule. Trypanosomes were present in the intertubular tissues of seminiferous tubules, and they occasionally crossed the myoid layer but never the basal lamina of seminiferous tubules. Inflammatory and other cells in the interstitium consisted of lymphocytes (35.2%), Leydig's cells (25.1%), macrophages (18.1%), plasma cells (12.3%), neutrophils (5.6%), eosinophils (3.2%), and mast cells (0.5%). Inflammatory cells including lymphocytes (51.6%), macrophages (43.9%), plasma cells (3.0%), and eosinophils (1.5%) occasionally breached the myoid layer and occupied the space between it and the basal lamina of the seminiferous tubule. Leydig's cells usually existed in clusters, had decreased mitochondrial size and secretory granules, and folding of the nuclear membrane.

Varicose axons bearing "synaptic" vesicles on the basal lamina of the human seminiferous tubules.

Normal (infant and adult) and pathological testes were examined by electron microscopy in order to study testicular innervation. Nerves composed of non-myelinated fibres were abundant in the tunica vasculosa of the tunica albuginea. These nerves penetrated into the testicular septa reaching the interstitial tissue. This showed numerous non-myelinated nerve fibres running among the Leydig cells and blood vessels. Single axons or small groups of them, partially surrounded by Schwann cells, approached: 1) the Leydig cells, 2) the interstitial blood vessels, and 3) the seminiferous tubules. Single naked axons were also observed primarily in the proximity of the seminiferous tubules. These axons showed varicosities containing both small and large "synaptic" vesicles. The latter were less numerous and contained a central dense core. Small vesicles were agranular. Some varicose axons ran across the myofibroblast layer of the tunica propria reaching the basal lamina of the seminiferous tubules at the level of the Sertoli cells but not at the level of the spermatogonia. The intercellular space between Sertoli cell and axon membrane was about 150-200 nm.

精巣と腓腹筋にみられる実験的ラットビタミンE欠乏性病変とヒト老年性病変との比較病理学的研究

Vitamin E is well recognized to act as a very versatile reagent for facilitating fertilization. Recently, particular attention has been focussed on the fact that Vitamin deficient subjects maydisplay changes quite similar to those described in cases of aged subjects.From this view-point the author has investigated the testis and skeletal muscles of Vitamin E deficient rats and aged group, and has compared their histo-pathological changes.In the testicular tissue, the decrease of spermatocyte, sperm, and number of Leydig cells Were observed the hyalinosis of the basal lamina of seminiferous tubules Was very striking. Moreover, proliferation of multinuclear giant cells which might be derived from spermatocytesand reserved Sertori cells were not unfrequently observed.It is noteworthy that hyaline degenerationof the basal lamina of tubules in the aged group constitute one of the conspicuousfeatures.In the m. gastrocnemius of experimental rats, uneven atrophic and degenerative changeswere observed associated with occasional floccular degeneration, increase of number of nuclei, non-reactive necrotic change, and proliferation of reticulin fibers. The muscular degenerationand atrophy are justifiable as the process resembling the dystrophic changes of the muscle, andobviously different from the neurogenic atrophy of the muscle which is documented as the groupatrophy of muscles.There are many similarities between Vitamin E deficiency changes and that of aging. Vitamin E is regarded as an anti-oxidative agent, and its deficiency may lead to peroxidation of thecytomembrane which eventually may cause extensive degenerative changes of the cell.From the above, the author has concluded that further detailed investigation of the biologicalaction of Vitamin E on various tissue injury and aging, may provide a clue to the understandingof the phenomenon of aging.