Lumen Of Seminiferous Tubules Research Articles

Mutation of the Thap4 gene causes dwarfism and testicular anomalies in rats and mice.

The petit (pet) locus is associated with dwarfism, testicular anomalies, severe thymic hypoplasia, and high postnatal lethality, which are inherited in autosomal recessive mode of inheritance in rats with a Wistar strain genetic background. Linkage analysis localized the pet locus between 98.7Mb and 101.2Mb on rat chromosome 9. Nucleotide sequence analysis identified 2bp deletion in exon 2 of the Thap4 gene as the causative mutation for pet. This deletion causes a frameshift and premature termination codon, resulting in a truncated THAP4 protein lacking approximately two-thirds of the C-terminal side. Thap4 is expressed in various organs, including the testis and thymus in rats. To elucidate the biological function of THAP4 in other species, we generated Thap4 knockout mice lacking exon 2 of the Thap4 gene through genome editing. Thap4 knockout mice also exhibited dwarfism and small testis but did not show high postnatal lethality. Thymus weights of adult Thap4 knockout male mice were significantly higher compared to wild-type male mice. Although Thap4 knockout male mice were fertile, their testis contained seminiferous tubules with spermatogenesis and degenerative seminiferous tubules lacking germ cells. Additionally, we observed vacuoles in seminiferous tubules, and clusters of cells in the lumen in seminiferous tubules in Thap4 knockout male mice. These results demonstrate that spontaneous mutation of Thap4 gene in rats and knockout of Thap4 gene in mice both cause dwarfism and testicular anomalies. Thap4 gene in rats and mice is essential for normal testicular development, maintaining spermatogenesis throughout the entire region of seminiferous tubules.

Antifertility potential of the extracts of <i>Aloe barbadensis</i> Miller on the testes and reproductive hormones of male rats

There is a plethora of effective female contraceptives with varied efficacy, usability, and acceptability levels but not for males, who have condoms with a high failure rate and vasectomy with poor reversibility. This study was to determine the antifertility potential of the extracts of Aloe barbadensis as a herbal contraceptive for males. Gel and aqueous leaf extracts of Aloe barbadensis were administered orally to thirty-five sexually matured albino male rats for 30 days, to determine their effects on the reproductive hormones and histopathology of the testes. The animals were divided into 7 groups A, B, C, D, E, F, and G. Each group was for a treatment regimen having 5 rats. Group A, the Control, was given a normal ration and water. Groups B, C, and D were administered 10mg/kg, 30mg/kg, and 70mg/kg of fresh A. barbadensis gel daily for 30 days while groups E, F, and G were administered 10mg/kg, 30mg/kg, and 70mg/kg of aqueous leaf extract respectively. A significant reduction in testosterone levels (p<0.05), insignificant reduction in follicle-stimulating hormone levels, and an increase in luteinizing hormone levels that were also not significant (p>0.05) were observed. Histopathological alterations of disrupted cytoarchitecture, absence of Leydig cells, and scanty spermatozoa in the lumen of seminiferous tubules were observed in the tissues of the testes of the treated groups of rats, and as such the extracts may be possible effective natural antifertility agents for males although, toxic constituents of the plant extracts should be screened out before it is allowed for consumption.

Expression of NELL2/NICOL-ROS1 lumicrine signaling-related molecules in the human male reproductive tract.

The maturation of spermatozoa is a regulated process, influenced by genes expressing essential secreted proteins in the proximal epididymis. Recent genetic studies in rodents have identified the non-sex steroidal molecular signals that regulate gene expression in the proximal epididymis. Germ cells in the testis secrete ligand proteins into the seminiferous tubule lumen The ligand proteins travel through the male reproductive tract lumen to the epididymis, where they bind to receptors, triggering the differentiation of the luminal epithelium for sperm maturation. It is, however, not fully unveiled if such a testis-epididymis trans-luminal signaling mechanism exists in other species, especially humans. In the present study, the rodent-type testis-epididymis trans-luminal signaling in the human male reproductive tract was evaluated in a step-by-step manner by analyzing testis and epididymis gene expression and signaling mediator protein function. There was a significant correlation between the epididymal expressions of mouse genes upregulated by the trans-luminal signaling and those of their human orthologs, as evaluated by the correlation coefficient of 0.604. The transcript expression of NELL2 and NICOL encoding putative ligand proteins was also observed in human testicular cells. In vitro experiments demonstrated that purified recombinant human NELL2 and NICOL formed a molecular complex with similar properties to rodent proteins, which was evaluated by a dissociation equilibrium constant of 110 nM. Recombinant human NELL2 also specifically bound to its putative receptor human ROS1 in vitro. Collectively, these findings suggest that the rodent-type testis-epididymis secreted signaling mechanism is also possible in the human male reproductive tract.

Fucoidan from Cladosiphon okamuranus enhances antioxidant activity and prevents reproductive dysfunction in polystyrene microplastic-induced male rats

Plastic pollution, including microplastic, has emerged as a severe environmental and public health problem. The health risks, especially in the case of reproductive damage caused by polystyrene microplastic (PS-MP) exposure, are emerging problems that need to be solved. This study aimed to investigate the effects of fucoidan extracted from Cladosiphon okamuranus on the polystyrene microplastic-induced oxidative stress of the Leydig (LC540) cells and reproductive damage in male rats. The oxidative stress of the LC540 cells and reproductive damage in the rats were induced by PS-MP. The fucoidan treatment reduces nitric oxide (NO) and reactive oxygen species generation in the LC540 cells. In the animal study, fucoidan treatment enhanced enzymatic antioxidant activities (glutathione peroxidase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase) and reduced malondialdehyde and nitric oxide production. Fucoidan supplementation also downregulates tumor necrosis factor-alpha, interleukin-6, and caspase-3 expression. Additionally, fucoidan upregulates testosterone levels, prevents the reduction of epithelium thickness, and reduces the area of the seminiferous tubule lumen. According to these conditions, fucoidan from Cladosiphon okamuranus prevents reproductive damage by downregulating oxidative stress and pro-inflammatory cytokines. Therefore, fucoidan can be used as a source of food supplements or functional food ingredients for reproductive or testicular damage management.

Physicochemical Assessment of Zinc Oxide Nanoparticle and Moringa oleifera Supplementation on the Male Reproductive System of a Diabetic Rat Model

The probable synergistic effect of chemically synthesized zinc oxide nanoparticles and locally secured Moringa oliefera was assessed on diabetic rats’ model along with a potential effect on the male reproductive system and rat male serological markers. The plant M. oliefera was procured from the University of Agriculture Faisalabad step solvothermal method was utilized to synthesize the zinc oxide nanoparticles. The zeta sizer, UV–vis, and spectroscopic analysis of ZnO nanoparticles were undertaken. A total of 50 healthy male albino Wistar rats weighing 250 ± 10 g were used in the experimental research study having were divided into six stops designated as negative control, positive control (PC), standard control (Std. C), ZnO nanoparticle group (ZnO), M. oliefera Group (MO), and a group of combinations thereof (ZnONP + MO). The mean body weight was observed to be significantly normalized in group ZnONP + MO, i.e., 230 ± 6 g in contrast to PC, i.e., 162 ± 4 g; P ≤ 0.05 . The ZnONP + MO combination had a normoglycemic effect, i.e., 154.4 ± 4.5 mg/dl as in opposition to the PC, i.e., 315.7 ± 3 mg/dl. The serum level of rat testosterone in the ZnONP + MO group was observed at 0.958 ± 0.08 ng/ml in opposition to PC, which revealed it at 0.442 ± 0.02 ng/ml. The follicle-stimulating hormone level in the ZnONP + MO group was recorded at 10.04 ± 0.04 mlU/ml significantly varied from the PC, whose level was noted at 5.08 ± 0.09 mlU/ml. The level of LH in the ZnONP + MO group was observed at 6.89 ± 0.08 mIU/ml, significantly different from PC at 3.78 ± 0.08 mIU/ml. Histopathological changes in the rat testes treated with alloxan alone revealed the distortion in the epithelium of seminiferous tubules (H&E stain). However, the histopathology of testes isolated from rats treated with the zinc oxide nanoparticle and M. oliefera combination showed almost normal spermatogenic activity, the lumen of seminiferous tubules contained sperms, normal spermatids, and outer epithelium layer of seminiferous tubules was intact.

Histomorphometric study of seminiferous tubules and epididymis in adult ram

This research set out to examine the epididymis and seminiferous tubules of adult male Rams, use both histological and histomorphometric methods. For this purpose, ten paired of testis of healthy adult local rams were taken from local abattoir in Fallujah. Tissue was removed and cut into tiny pieces from different parts of the testicles before being put in Bouin's solution. The tissue samples were processed by common paraffin technique then stained with routine H & E stain as well as Masson's trichrome stain. According to the results of the present histological analysis, the ram's testis is enclosed by a capsule of thick irregular connective tissue, and its lobules are separated by trabecular and interlobular septa, with seminiferous tubules and interstitial tissue making up each lobule. Spermatogenic cells and Sertoli cells border the seminiferous tubules. Spermatozoa develop from spermatogenic cells. Spermatogonia are the earliest stages of spermatogenic cells; they are tiny, spherical cells with black, spherical nuclei that sit on the basement membrane. Primary spermatocytes, which are bigger cells with often-distinct chromatin, are generated during mitosis in the spermotogonia. Because of their rapid second meiotic division and subsequent formation of haploid spermatids, secondary spermatocytes are seldom seen. Clustered near the seminiferous tubule's lumen, the spermatids are spherical cells with pale nuclei. There are less Sertoli cells (sustentacular cells) than spermatogenic cells. The nucleus is oval or triangular in shape and pale in colour, with a large nucleolus. Leydig cells are found in the connective tissue between neighbouring tubules. Epithelium that is pseudostratified lines the epididymis. The stereocilia are long cytoplasmic structures that extend into the lumen. Circular smooth muscle is present in varying amounts, and the epithelium is supported by a connective tissue lamina propria.In conclusion, the current study concluded that the thickness of the epithelium of epididymis was decrease toward the tail while the thickness of the smooth muscle layer was increase toward the tail.

Evaluation of chemical castration using intra-testicular injection of zinc gluconate into the testis of the male donkey versus surgical castration: antimullerian hormone as an endpoint marker

BackgroundChemical castration of male animals is an alternative to surgical castration for inducing azoospermia, consequent sterility. Intra-testicular injection of zinc gluconate has been used for chemical castration in several animal species. However, its application to equine species, such as donkeys, has yet to be reported. This study aimed to evaluate the use of zinc gluconate for the chemical castration of male donkeys and to compare its effectiveness relative to routine surgical castration. For this purpose, investigations of serum testosterone and anti-Müllerian hormone levels, testicular ultrasonographic echogenicity, and histopathological findings were performed.MethodsFourteen clinically healthy adult male donkeys were randomly and equally divided into two groups. The donkeys in group I (n = 7) underwent surgical castration. The donkeys in group II (n = 7) received intra-testicular zinc gluconate injections. The donkeys were kept under close clinical observation for 60 days. Abnormalities in donkey behavior and gross alterations in the external genitalia were recorded daily. Serum testosterone and anti-Müllerian hormone (AMH) levels were measured 15 days before the start of the treatment and 15, 30, 45, and 60 days after treatment. The testicles of group II donkeys were evaluated ultrasonographically. At the end of the study, the testes were removed and histologically examined.ResultsSerum testosterone levels significantly declined compared to pre-castration levels in surgically castrated donkeys (group I), but donkeys exposed to chemical castration (group II) showed a non-significant reduction in testosterone levels. Donkeys in the surgical group had considerably lower serum AMH levels. In contrast, there was a non-significant (p > 0.05) increase in AMH levels in the chemical group compared with the pre-sterilization level. In addition, ultrasonographic examination revealed that the testicular echo-density had changed, as observed by a few scattered hyperechoic regions throughout the entire testis parenchyma. The histopathological investigation confirmed the presence of necrosis of the spermatogenic epithelium, increased thickness of the basement membrane of the seminiferous tubules, marked interstitial fibrosis, and shrinkage of the seminiferous tubules. Furthermore, syncytial giant cells were present in the lumen of seminiferous tubules and were associated with Sertoli cell vacuolation. Donkeys subjected to chemical castration (group II) had orchitis, as confirmed histopathologically.ConclusionIntra-testicular injection of zinc gluconate resulted in histopathological and ultrasonographic testicular changes in adult male donkeys, which may affect their reproductive potential. However, it did not significantly alter serum testosterone or AMH levels, indicating that it cannot be used as a substitute for surgical castration in male donkeys.

Immunohistochemical and Ultrastructural Evaluation of Spermatogenic Alteration by P53 under the Influence of Bisphenol-A

The p53 gene is a tumor suppressor gene and, thus, plays an important role in cell cycle, cell senescence, DNA repair, and cell death. Since testicular tissues go through a continuous and complex process of spermatogenesis, p53 is likely to play a significant role in the regulation of germ cell proliferation and spermiogenesis. In the present study, the specific localization of p53 in testicular tissues was determined by comparing it with BPA induced toxicity. Four groups containing 10 albino rats each were designated as Group I: Control, Group II: 10 mg/kg BPA, Group III: 50 mg/kg BPA, and Group IV: 100 mg/kg BPA. Daily administration of BPA was carried out through oral gavage for 6 weeks by dissolving the assigned weight of BPA in olive oil. Testicular tissues were investigated for expression of p53 by immunohistochemistry, and testicular sperms were examined under a scanning electron microscope. Results showed that p53 was exclusively expressed in the spermatogonia of animals exposed to 10 mg/kg BPA. The highest expression of p53 was present in animals exposed to 50 mg/kg BPA; besides spermatogonia, spermatocytes and spermatids also indicated positive expression. However, relatively lower expression was evident in animals exposed to 100 mg/kg BPA, as most cellular architecture was already distorted significantly, and germ cells appeared to have fallen into the lumen of seminiferous tubules. The ultrastructure of testicular sperm indicated specific damage to the perforatorium, plasma membrane, and connecting pieces around the neck, and tail. Damages occurring in the head cap segment of the perforatorium indicated an alteration during spermiogenesis. In conclusion, it is highly likely that a BPA induced alteration in the expression of p53 may have affected spermiogenesis through spermatogenesis.

In Vitro and In Vivo Models for Drug Transport Across the Blood-Testis Barrier.

The blood-testis barrier (BTB) is a selectively permeable membrane barrier formed by adjacent Sertoli cells (SCs) in the seminiferous tubules of the testes that develops intercellular junctional complexes to protect developing germ cells from external pressures. However, due to this inherent defense mechanism, the seminiferous tubule lumen can act as a pharmacological sanctuary site for latent viruses (e.g., Ebola, Zika) and cancers (e.g., leukemia). Therefore, it is critical to identify and evaluate BTB carrier-mediated drug delivery pathways to successfully treat these viruses and cancers. Many drugs are unable to effectively cross cell membranes without assistance from carrier proteins like transporters because they are large, polar, and often carry a charge at physiologic pH. SCs express transporters that selectively permit endogenous compounds, such as carnitine or nucleosides, across the BTB to support normal physiologic activity, although reproductive toxicants can also use these pathways, thereby circumventing the BTB. Certain xenobiotics, including select cancer therapeutics, antivirals, contraceptives, and environmental toxicants, are known to accumulate within the male genital tract and cause testicular toxicity; however, the transport pathways by which these compounds circumvent the BTB are largely unknown. Consequently, there is a need to identify the clinically relevant BTB transport pathways in in vitro and in vivo BTB models that recapitulate human pharmacokinetics and pharmacodynamics for these xenobiotics. This review summarizes the various in vitro and in vivo models of the BTB reported in the literature and highlights the strengths and weaknesses of certain models for drug disposition studies. SIGNIFICANCE STATEMENT: Drug disposition to the testes is influenced by the physical, physiological, and immunological components of the blood-testis barrier (BTB). But many compounds are known to cross the BTB by transporters, resulting in pharmacological and/or toxicological effects in the testes. Therefore, models that assess drug transport across the human BTB must adequately account for these confounding factors. This review identifies and discusses the benefits and limitations of various in vitro and in vivo BTB models for preclinical drug disposition studies.

Effects of aqueous methanolic extract of Bridella ferruginea stem bark on the spermatogenesis, accessory reproductive organs, and organosomatic indices in pubertal male Rattus Norvegicus

There is an increasing trend in the usage of medicinal plants in livestock production with some significantly toxic effects especially to vital organs such as reproductive tissues which necessitates this search. Twenty (20) matured male rats weighing 150-239g randomly assigned into 4 groups (n=5) were used for the sub-acute toxicity. Group 1 rats were treated with distilled water while groups 2-4 were treated with 100, 200, and 400mg/kg respectively of the aqueous methanolic extract of Bridelia ferruginea stem bark (AMEBFSB) for 30 days. Animals were terminated on day 31 and some of the reproductive organs were collected and weighed. The Cauda epididymis served as a semen source for sperm analysis and testes were collected for histological examination. There was a significant decrease (p<0.05) in the final body weights, average weights of testes, epididymis, vas deferens, seminal vesicles, and prostate gland of the treated groups as compared to the control. The semen evaluated showed a significant decrease (p<0.05) in sperm motility, sperm count, and percent normal sperm cell morphology in the 400mg/kg group, no significant difference (p<0.05) in the sperm viability and acrosome integrity as compared to control. Photomicrograph revealed loss and thinning of germinal epithelium and widened seminiferous tubule lumen. In conclusion, the oral administration of AMEBFSB for 30 days in male rats showed that the plant has antispermatogenic effects on reproductive functions.

Testicular toxicity in mice exposed to terephthalic acid in utero and during lactation.

Terephthalic acid (TPA) is a worldwide aromatic compound widely used to manufacture resins and the raw material for the polymerization reaction with ethylene glycol to produce polyethylene terephthalate, known as PET. The use of TPA extends to the synthesis of phthalates, plasticizers used in various industrialized products such as toys and cosmetics. The present study aimed to evaluate the testicular toxicity of terephthalic acid on male mice exposed in utero and during lactation to TPA in different developmental windows. The animals were treated intragastric with TPA at stock dispersal dosages corresponding to 0.0014g/ml and 0.56g/ml of TPA in 0.5% v/v carboxymethylcellulose as well as the control dose, composed solely of dispersion of carboxymethylcellulose (0.5% v/v). Four experimental windows were established: group I-treatment in utero, in the fetal period (gestational day-GD 10.5-18.5), with euthanasia at GD 18.5; group II-treatment in utero, in the fetal period (GD 10.5-18.5) and the lactational period (postnatal day (PND-15)), with euthanasia at 15days; group III-treatment in utero in the fetal period (DG 10.5-18.5) with euthanasia at 70days (age of sexual maturity, PND 70); group IV-treatment in utero, in the fetal period (GD 10.5-18.5) and the lactational period (PND-15), with euthanasia at 70days (PND70). The results indicate that TPA changes the reproductive parameters (testicular weight, GI, penis size, and anogenital index) only at the dose of 0.56g/ml in the fetal period. Data on the volumetric ratio of the testis elements show that the dispersion with the highest concentration of TPA significantly altered the blood vessel/capillary, lymphatic vessel, and connective tissue percentages. Only at the dose of 0.56g/ml TPA was it effective in decreasing the Leydig and Sertoli cell numbers of the euthanized animals at GD 18.5. In group II, TPA increased the diameter and lumen of seminiferous tubules, which indicates that TPA accelerated the maturation process of Sertoli cells without changing the number and the nuclear volume of these cells. The Sertoli and Leydig cell numbers of the 70-day animals exposed to TPA in the gestational and lactational period were similar to the control. Therefore, the present study is the first in the literature to show that TPA presents a testicular toxicity during fetal (DG18.5) and postnatal life (PND15), without repercussion in adulthood (70days).

Disrupted intercellular bridges and spermatogenesis in fatty acyl-CoA reductase 1 knockout mice: A new model ofether lipid deficiency.

Peroxisomal fatty acyl-CoA reductase 1 (FAR1) is a rate-limiting enzyme for ether lipid (EL) synthesis. Gene mutations in FAR1 cause a rare human disease. Furthermore, altered EL homeostasis has also been associated with various prevalent human diseases. Despite their importance in human health, the exact cellular functions of FAR1 and EL are not well-understood. Here, we report the generation and initial characterization of the first Far1 knockout (KO) mouse model. Far1 KO mice were subviable and displayed growth retardation. The adult KO male mice had smaller testes and were infertile. H&E and immunofluorescent staining showed fewer germ cells in seminiferous tubules. Round spermatids were present but no elongated spermatids or spermatozoa were observed, suggesting a spermatogenesis arrest at this stage. Large multi-nucleated giant cells (MGC) were found lining the lumen of seminiferous tubules with many of them undergoing apoptosis. The immunofluorescent signal of TEX14, an essential component of intercellular bridges (ICB) between developing germ cells, was greatly reduced and mislocalized in KO testis, suggesting the disrupted ICBs as an underlying cause of MGC formation. Integrative analysis of our total testis RNA-sequencing results and published single-cell RNA-sequencing data unveiled cell type-specific molecular alterations underlying the spermatogenesis arrest. Many genes essential for late germ cell development showed dramatic downregulation, whereas genes essential for extracellular matrix dynamics and cell-cell interactions were among the most upregulated genes. Together, this work identified the cell type-specific requirement of ELs in spermatogenesis and suggested a critical role of Far1/ELs in the formation/maintenance of ICB during meiosis.

Morphology of the male rat gonad after in Utero exposure to immunosuppressants

The protocol for immunosuppression of pregnant women is based on immunosuppressant panels. The aim of the study was to determine the influence of commonly applied combinations of immunosuppressants to pregnant rats on the morphology of the offspring’ testes. Pregnant rats were treated with cyclosporin A (CsA), mycophenolate mofetil (MMF) and prednisone (Pred) (CMG); tacrolimus (Tc), MMF and Pred (TMG); CsA, everolimus (Ev) and Pred (CEG). Testes of mature offspring underwent morphological analysis. Mainly in the testes of CMG and TMG rats the morphological and functional changes were observed: immature germ cells (GCs) in the seminiferous tubule (ST) lumen, invaginations of the basement membrane, infolding to the seminiferous epithelium (SE), the ST wall thickening, increased acidophilia of Sertoli cells’ (SCs) cytoplasm, large residual bodies near the lumen, dystrophic ST and tubules look like the Sertoli cell-only syndrome, Leydig cells with abnormal cell nucleus, hypertrophy of the interstitium, blurring of the boundary between ST wall and interstitium, a reduced number of GCs in the SE, vacuolation of the SE. In the CEG there were only a reduced number of GCs in some tubules and vacuolization of SCs. The safest combination of drugs was CEG, while the TMG and CMG were gonadotoxic.

A Comparative Study of the Cytohistomorphological and Biochemical Effect of Alcoholic Beverages (Brandy, Beer, Sour Wine, Dry Gin) on the Testes of Adult Male Wistar Rats

This study was designed to evaluate the cytohistomorphological and biochemical effect of alcoholic beverages on the prostate gland of adult male Wistar rats. Sixty-five (65) rats weighing between 180-230g were used for this experiment. They were randomly divided into 13 groups of five (5) animals each. Group 1 was the normal control. Group 2-13 were the experiment groups. Group 2, 3 and 4 were treated with 1.23mg/kg, 2.45mg/kg and 3.68mg/kg bodyweight of brandy respectively. Group 5, 6 and 7 were treated with 17.32mg/kg, 34.64mg/kg and 51.96mg/kg body weight of beer respectively. Group 8, 9 and 10 were treated with 12.25mg/kg, 24.96mg/kg and 36.74mg/kg bodyweight of soured wine respectively. Group 11, 12 and 13 were treated with 1.73mg/kg, 3.46mg/kg and 5.20mg/kg bodyweight of dry gin respectively. Administration was done daily for 28 days and orally using orogastric tube. On the 29th day, the animals were sacrificed using chloroform inhalation anaesthesia. The blood samples were aspirated via cardiac puncture and centrifuged for biochemical analysis, and testicular tissues were harvested, fixed in 10% buffered formalin, processed, and stained with haematoxylin and eosin. Body weight showed significant (p<0.05) increase in brandy administered groups compared to control. For testicular weight, there was an insignificant increase in all the treated groups except the group administered with 3.69mg/kg which showed insignificant decrease compared to control. Results for TT showed a general significant (p<0.001) decrease in all administered groups compared to control. FSH showed significant (p<0.05;0.01;0.001) decrease in group administered 3.69mg/kg of brandy, all administered groups of soured wine and group administered 5.20mg/kg of dry gin compared to control. Histology showed narrowed and elongated lumen of seminiferous tubule, hypertrophied sertoli cells, destroyed interstitial cells of leydig, distorted seminiferous tubules with degenerating spermatogenic cells in the administered groups compared to control. In conclusion, alcohol beverages pose adverse effects on the testes. KEYWORDS: Alcoholic Beverages, Testes, Histology, Testosterone, FSH.

Ameliorative Potentials of Bombax costatum Pellegr. & Vuillet Chloroform Stem Bark Extract on Pentylenetetrazole-induced Testicular Damage in Wistar Rats

This study was conducted out to find out the ameliorative potentials of Bombax costatum Pellegr. & Vuillet chloroform extract (BCCE) on Pentylenetetrazole (PTZ)-induced testicular damage in albino Wistar rats. Eighteen adult rats were randomly allotted into three groups (n=6) after one week of acclimatization. Group one served as a control and was given normal saline. Group two (PTZ only) was given 35mgkg-1 body weight PTZ. Group three received BCCE 250mgkg-1 followed by PTZ 35mgkg-1, the treatments lasted for one week. The rats were euthanized and testicular tissues were processed for histological study, while blood samples were collected and used for the hormonal assay. The interstitial Leydig cells have been disrupted by the PTZ toxicity which resulted in a significant decrease (p<0.05) of testosterone in the PTZ-induced rats as compared with the control group, while there was no significant difference in group three that was given BCCE when compared to the control. Also, there was no significant difference in the luteinizing hormone (LH) and follicles stimulating hormones (FSH) levels in all the rat groups. The histological features showed that PTZ has a deleterious effect on testicular tissues with seminiferous tubule lumen devoid of spermatozoa. The histology of the group that received BCCE 250mgkg-1 shows the lumen of the seminiferous tubules with spermatozoa indicating that BCCE exerts a protective effect against PTZ-induced damage. This suggests that BCCE has an ameliorative effect against PTZ-induced testicular toxicity in rat models.

Determination of possible contraceptive potential of methanolic leaf extract of Mentha longifolia L. in adult male rats: a biochemical and histological study.

Mentha longifolia L (ML) is locally used as an herbal contraceptive. We aim to assess the toxicity profile and validate the antifertility effects of its leaves in male rats. Adult male Sprague Dawley rats (n=10 animals/group) were divided into four groups. The Control group received .9% saline, while groups II, III, and IV received 50, 75, and 100mg/kg/day ML doses for 28days. ML induced a dose-dependent decrease in percent fertility as well as the number of pups born in ML preexposed pairs, with maximum change observed in the ML3 group (60%, 3.75±1.25) as compared to control (100%, 8.75±.48) group. A significant (P<.01) decrease in the rate of daily sperm production was observed in the ML3 group (.86±.16) in contrast to the control (2.93±.05). The levels of catalase and superoxide dismutase declined significantly in treatment groups (ML1-P<.001, ML2-P<.05, and ML3-P<.01). A significant increase in the production of reactive oxygen species (P<.001) was found in all treatment groups. The concentrations of LH (P<.001), FSH (P<.001), and testosterone (P<.001) were significantly reduced in a dose-dependent manner among ML-treated groups. Decreases in spermatogonial populations, mature spermatids, seminiferous tubule diameter, lumen diameter, and epithelial height were noticed at higher doses. The reversibility study depicted a reduction in both pregnancy outcomes and litter size. Besides contraceptive effects, M. longifolia L usage is associated with oxidative stress; therefore, it is important to consume this herb appropriately and its excessive usage should be prohibited.

Disturbance in Some Fertility Biomarkers Induced and Changes in Testis Architecture by Chronic Exposure to Various Dosages of Each of Nonylphenol or Bisphenol A and Their Mix.

Simple SummaryIn this study, 70 male albino rats were allocated to the control group (GI) and were given 1 mL of ethanol; G, II, and G III were treated with 100 mg/Kg of each of BPA and NP, G IV and G V were treated with 25 mg/Kg of each of BPA and NP, G VI was given a mixture of a high dose of BPA and NP, and G VII was given a mixture of a low dose of BPA and NP. A significant elevation in the TNF Alpha, TNF Beta, and Caspase 3 serum was recorded individually and in the groups treated with high doses. In conclusion, exposure to BPA and NP strongly impacts antioxidants, immune-inflammatory mediators, and testicular tissue architecture. Furthermore, the data from this investigation support the idea that exposure to BPA and NP in daily life has multiple damages.This investigation was conducted to demonstrate the potential impacts of different doses of Bisphenol A (BPA) or Nonylphenol (NP) and their mixtures on some biological activities in male albino rats. Seventy male albino rats were allocated to the control group (GI) and were given 1 mL of ethanol. G II and G III were given 100 mg/kg of each of BPA and NP, G IV and G V were given 25 mg/kg of each of BPA and NP, G VI was given a high dose of BPA and NP, and G VII was given a low dose of BPA and NP. All animals were treated orally for 60 days. Serum biomarkers of oxidative stress, antioxidants, immune-inflammatory mediators, and apoptotic markers were determined, as well as a histopathological examination of the testis at the end of the experimental period. The results obtained showed a pronounced increase in malondialdehyde (MDA), protein carbonyl (PC), and 4-hydroxynonenol (4-HNE), concomitant with a significant reduction in serum Superoxide dismutase (SOD), catalase enzyme (CAT), and total antioxidant capacity (TAC) in all treated groups. A significant elevation in TNF Alpha, TNF Beta, and Caspase 3 serum was recorded individually and in the groups treated with high doses. The disturbance is represented by histological damage in the testis in the germinal epithelium and a decrease in spermatozoa inside the lumen of seminiferous tubules. The effects on testis tissues were dose-dependent, pronounced in mixture doses, and remarkable in higher doses. In conclusion, exposure to BPA and NP strongly impacts antioxidants, immune-inflammatory mediators, and testis tissue architecture. Furthermore, the data from this investigation support the idea that exposure to BPA and NP in daily life has multiple damages.

Histopathological Changes in Rat Testes Induced by Potassium Bromate and Potential Ameliorative Effects of Ruta chalepensis Oil Extract

Potassium bromate (KBrO3) is used as a food additive mainly in the bread-making process. Ruta chalepensis (R. chalepensis) is an excellent and valuable bioactive plant that produces a range of complex flavonoids. The present study was aimed to investigate the possible protective and therapeutic effect of R. chalepensis against KBrO3 toxic effects on rat testis using histopathology investigation. Fifty adult male albino rats were used in the present study. The rats were divided into five groups each containing 10 rats. First group was kept as control, second group received oil extract of R. chalepensis, third group was treated with KBrO3, forth group (Protective group) and fifth group (Therapeutic group). All groups of animals were sacrificed at 2 and 4 weeks. The results of the present study showed that the administration of R. chalepensis caused disarrangement of spermatogonia throughout the lumen of seminiferous tubule. Additionally, abnormal widening of interstitial spaces with degeneration of interstitial cells. There was necrosis to the spermatocytes with inhibition of the spermatogenic process. KBrO3 group showed necrosis of seminiferous tubule, loss in some spermatogenic cells. There were fibrous stroma between the seminiferous tubules. Also revealed vacuolation, edematous, hyalinization and loss of interstitial connective tissue cell with haemorrhage between seminiferous tubules. Protective group showed seminiferous tubule atrophy, disrupted germ cell layers and disappearance of Leydig cells. The therapeutic group, showed interstitial hemorrhage, with necrosis in spermatogonia and vacuolation. It may be concluded that KBrO3 is toxic to testis and R. chalepensis at the tested dose is not beneficial as protective and curative agent.

Dose dependent intra-testicular accumulation of silver nanoparticles triggers morphometric changes in seminiferous tubules and Leydig cells and changes the structural integrity of spermatozoa chromatin

Citrate-coated silver nanoparticles were synthesized in one step method using tri-sodium citrate and silver nitrate at pH 6.1. After synthesis, the resulting silver nano-suspension was characterized using UV–visible spectroscopy, dynamic light scattering, high resolution-scanning electron microscopy, energy dispersive x-ray spectroscopy and fourier transform infrared-spectroscopy. The particles were intraperitoneally injected into Swiss albino male mice for a period of one complete spermatogenic cycle. The LD50 was determined following the procedure of Dixon's Up-and-Down method. The intra-testicular level of silver was measured using the technique of inductively coupled plasma-mass spectrometry. The intra-testicular location of accumulated nanoparticles was observed using auto-metallography. The cytology and volume of Leydig cells were assessed and analysed. Following the exposure to silver nanoparticles, it was found that accumulation of nanoparticles inside the seminiferous tubules is a dose-dependent process. The deposition of silver agglomerate induced morphometric changes in the lumen of seminiferous tubules and Leydig cells. The exposure also caused significant changes at the level of structural integrity of sperm chromatin material and variable damages to sperm DNA.

Sertoli cells require TDP-43 to support spermatogenesis†.

TAR DNA binding protein of 43 kD (TDP-43) is an evolutionarily conserved, ubiquitously expressed transcription factor and RNA-binding protein with major human health relevance. TDP-43 is present in Sertoli and germ cells of the testis and is aberrantly expressed in the sperm of infertile men. Sertoli cells play a key role in spermatogenesis by offering physical and nutritional support to male germ cells. The current study investigated the requirement of TDP-43 in Sertoli cells. Conditional knockout (cKO) of TDP-43 in mouse Sertoli cells caused failure of spermatogenesis and male subfertility. The cKO mice showed decreased testis weight, and low sperm count. Testis showed loss of germ cell layers, presence of vacuoles, and sloughing of round spermatids, suggesting loss of contact with Sertoli cells. Using a biotin tracer, we found that the blood-testis barrier (BTB) was disrupted as early as postnatal day 24 and worsened in adult cKO mice. We noted aberrant expression of the junction proteins connexin-43 (gap junction) and N-cadherin (ectoplasmic specialization). Oil Red O staining showed a decrease in lipid droplets (phagocytic function) in tubule cross-sections, Sertoli cells cytoplasm, and in the lumen of seminiferous tubules of cKO mice. Finally, qRT-PCR showed upregulation of genes involved in the formation and/or maintenance of Sertoli cell junctions as well as in the phagocytic pathway. Sertoli cells require TDP-43 for germ cell attachment, formation and maintenance of BTB, and phagocytic function, thus indicating an essential role for TDP-43 in the maintenance of spermatogenesis.