Structure Of Seminiferous Tubules Research Articles

Histological and Immunohistochemical Examination in Testicular Tissue of Diabetic Rats Treated with Dandelion (Taraxacum officinale) Extract

The aim of this study was to evaluate the changes induced in testicular tissue by administration of dandelion (Taraxacum officinale) extract and its effects on the release of enzymatic antioxidants SOD-2 by immunohistochemical methods in rats with experimental diabetes. Degenerative changes in the seminiferous tubule structure were observed in the testicular sections of the diabetes group. Separations between the basement membrane and the seminiferous epithelium, disruptions in the structure and localization of spermatogenic cells toward the lumen of the seminiferous tubules, thickening of the basement membrane and tunica albuginea, changes in the shape of Sertoli cells, and shrinkage of the nucleus of Leydig cells were observed. While no immunopositivity was observed in spermatogonium and primary spermatocytes in the diabetes group, weak SOD-2 immunoreactivity was observed in other cells of the spermatogenic series and moderate SOD-2 immunoreactivity was observed in the diabetes+dandelion group. In conclu-sion, it was found that histopathological changes occurred in the testicular tissue and SOD-2 immunoreactivity decreased in diabetes, a chronic metabolic disease. It was concluded that dandelion administration may have beneficial effects on decreased SOD-2 immunoreactivity.

Filtered and Unfiltered Cigarette Effect on White Rats Male Reproduction (Rattus norvegicus L.)

Cigarette smoke can cause various health problems, one of which is a decrease in fertility in males. The purpose of this study was to determine the effect of smoke exposure between with and without filter hand-rolled kretek cigarettes (SKT) and white cigarettes on white rats male reproduction: sperm quality in the form of sperm motility and viability, also the structure of seminiferous tubules. This study used 5 treatment groups (KP), namely controls not given exposure to cigarette smoke (KP0), exposure to unfiltered kretek cigarette smoke (KP1), exposure to filtered kretek cigarettes (KP2), exposure to unfiltered white cigarette smoke (KP3) and exposure to secondhand smoke. filtered white cigarettes (KP4) with treatment placement arrangement in a Completely Randomized Design (CRD) and if there is a significant difference, a Duncan Multiple Range Test (DMRT) is carried out. The results obtained on sperm quality, namely sperm motility and viability, decreased. Good quality at KP0 then the quality decreases successively starting from KP1, KP2, KP3 and KP4. The results on histpathological observations of the seminiferous tubules, there were no pathological changes found in all treatments. In contrast to the histopathology of seminiferous tubules, the number of spermatogenic cells showed a significant difference between treatments with the least average spermatogenic count and a decrease in the number of cells from spermatogonia cells to spermatid cells, namely in the treatment group exposure to filtered white cigarette smoke (KP4). Overall, the exposure that caused the most severe impact on all measured parameters was in the treatment group of filtered white cigarette smoke (KP4).

Effects of the adsorption behavior of polyamide microplastics on male reproductive health by reduction of testosterone bioavailability

Microplastics (MPs) are global environmental pollutants with potential toxicity concerns, and their effects on the reproductive system have attracted increasing attention. This study investigated the interaction between MPs and mammalian biomolecules, focusing on the relationship between the testosterone adsorption behavior of MPs and male reproductive health. The adsorption capacity of different types of MPs for testosterone was evaluated in vitro experiments. Polyamide (PA)-MPs exhibited stronger adsorption, while polymethyl methacrylate (PMMA)-MPs displayed the weakest adsorption. Sorption equilibrium between PA-MPs and testosterone was achieved within 6 h, fitting the Pseudo-2nd-order model and Langmuir isotherm. The effects of MPs on male reproduction in mice was determined in vivo experiments. Male mice were treated with 0.1 and 0.5 mg/d PA-MPs/PMMA-MPs by gavage once per day for 28 days. The results showed that only 0.5 mg/d PA-MP exposure induced decreased serum testosterone levels, increased testicular testosterone levels compared to the control, and more severe damage to seminiferous tubule structure, sperm motility and sperm morphology compared to the PMMA-MPs group. Meanwhile, PA-MPs could reduce intracellular nuclear translocation of androgen receptor (AR) mediated by testosterone, while PMMA-MPs had no impact. The study revealed that PA-MP adsorption reduced testosterone bioavailability and caused sperm quality to decline, offering new insights into the combined toxicity mechanism of MPs in male mammals.

UCHL1 maintains microenvironmental homeostasis in goat germline stem cells.

Spermatogonial stem cells (SSCs) play a crucial role in mammalian spermatogenesis and maintain the stable inheritance of the germline in livestock. However, stress and bacterial or viral infections can disrupt immune homeostasis of the testes, thereby leading to spermatogenesis destruction and infertility, which severely affects the health and productivity of mammals. This study aimed to explore the effect of ubiquitin C-terminal hydrolase L1 (UCHL1) knockdown (KD) in goat SSCs and mouse testes and investigate the potential anti-inflammatory function of UCHL1 in a poly(I:C)-induced inflammation model to maintain microenvironmental homeostasis. Invitro, the downregulation of UCHL1 (UCHL1 KD) in goat SSCs increased the expression levels of apoptosis and inflammatory factors and inhibited the self-renewal and proliferation of SSCs. Invivo, the structure of seminiferous tubules and spermatogenic cells was disrupted after UCHL1 KD, and the expression levels of apoptosis- and inflammation-related proteins were significantly upregulated. Furthermore, UCHL1 inhibited the TLR3/TBK1/IRF3 pathway to resist poly(I:C)-induced inflammation in SSCs by antagonizing HSPA8 and thus maintaining SSC autoimmune homeostasis. Most importantly, the results of this study showed that UCHL1 maintained immune homeostasis of SSCs and spermatogenesis. UCHL1 KD not only inhibited the self-renewal and proliferation of goat SSCs and spermatogenesis but was also involved in the inflammatory response of goat SSCs. Additionally, UCHL1 has an antiviral function in SSCs by antagonizing HSPA8, which provides an important basis for exploring the specific mechanisms of UCHL1 in goat spermatogenesis.

BDE-209 disturbed proliferation and differentiation of spermatogonia during mitotic process through estrogen receptor α.

Deca-bromodiphenyl ether (BDE-209) exposure caused spermatogenesis disorder resulting in poor sperm quality has become a public concern in recent years. Spermatogenesis refers to the process by which the division of spermatogonia stem cells (SSCs) produces haploid spermatozoa, including mitosis, meiosis, and spermiogenesis. However, the mechanism of mitosis including proliferation and differentiation of spermatogonia dysfunction induced by BDE-209 remains largely unclear. Here, our data showed that BDE-209 exposure caused a decline in sperm quality with seminiferous tubule structure disorder in rats. In addition, BDE-209 exposure damage spermatogonia proliferation and differentiation with decreasing level of PLZF and cKit in testis. Moreover, rats exposed to BDE-209 decreased the expression of ERα, whereas an elevated expression of Wnt3a and Wnt5a. Mechanistically, supplementation with propipyrazole triol (PPT, a selective ERα pathway agonist) rescued sperm quality and attenuated impairment of proliferation and differentiation of spermatogonia in BDE-209-induced rats. Therefore, ERα plays a crucial role in the proliferation and differentiation of spermatogonia during mitotic process. In conclusion, our study clarified the role of ERα in BDE-209-induced spermatogonia proliferation and differentiation in rats and provides a potential therapeutic application on poor sperm quality caused by BDE-209 exposure.

Exosomes obtained from adipose mesenchymal stem cells prevent ischemia-reperfusion injury after torsion-detorsion in rat testes.

To investigate the effect of exosomes obtainedfrom adipose-derived mesenchymal stem cells (AD-MSCs) on testicular ischemia-reperfusion (I/R) injury. AD-MSCs from rat adipose tissue were cultured. Characterization of cells was evaluated with CD44, CD90, CD34 and CD45 antibodies. Exosomes from AD-MSCs were obtained with the miRCURYexosomeisolation kit. 21 rats were divided into 3 groups. The I/R model was created as 720° torsion for 4h and reperfusion for 4h. In the Sham group (SG), only scrotal incision was made. 100µl of medium in the torsion-control group (T-CG) and 100µl of exosome in the treatment group (TG) were injected into the testicular parenchyma after detorsion. Johnsen scores of testicles were determined. Apoptosis was evaluated by the TUNEL method. It was observed that the seminiferous tubule structures were partially disrupted in T-CG, but normal in SG and TG. Johnsen scores in SG, T-CG, and TG were 8.64 ± 0.39, 7.71 ± 0.37, and 8.57 ± 0.39, respectively. Apoptotic cell distribution was 11.28 ± 5.25%, 60.58% ± 1.68% and 17.71 ± 8.34% in SG, T-CG and TG, respectively. In both parameters, the difference between SG and TG was insignificant (p > 0.05), the difference between T-CG/TG and SG/T-CG was significant (p < 0.05). Exosomes obtainedfromAD-MSCs are effective in preventing testicular I/R injury. This effect appears to occur because of suppression of apoptotic activity.

Pathophysiology, diagnosis, and management of testicular degeneration in the bull

Testicular degeneration in the bull is described as diffuse disintegration of seminiferous tubule structure and function causing a profound reduction in sperm concentration, scrotal circumference, and an increase in sperm morphologic abnormalities resulting in subfertility or permanent infertility. Degenerative changes within the seminiferous epithelium occur most commonly after an insult to spermatogenesis or may be idiopathic. Common insults include trauma, stress (environmental, heat, nutritional), or toxins. Idiopathic is most often associated with age-related changes. Clinical signs of testicular degeneration are discussed along with approaches to treatment and potential prognoses as related to treatment and management practices that can be employed by the general practitioner and specialist alike. This review will also encompass a discussion on the pathophysiology of this disease so that the practitioner has a detailed understanding of the disease, which improves their ability to employ evidence-based medicine and sound clinical reasoning.

A mosaic karyotype of 45,X/46,X,psu idic(Y)(q12) in a ten-year-old boy: integrating high-throughput sequencing with cytogenetic technique for precise diagnosis and genetic counselling

BackgroundIsodicentric Y chromosome (idic(Y)) is the most commonly reported aberration of the human Y chromosome, which is an important cause of abnormal sexual development. The breakpoints of isodicentric Y chromosome mostly occurred in Yq11.2 and Yp11.3, however, the breakpoints in Yq12 are relatively rare.Case presentationWe described a 10-year-old boy presenting hypospadias, micropenis and short stature, as well as unilateral cryptorchidism without normal testicular seminiferous tubules structure by biopsy. Whole exome sequencing didn’t find any pathogenic/likely pathogenic variants related to phenotypes of this patient. Copy number variation sequencing showed the duplication of whole Y chromosome. Subsequently, karyotyping and FISH analyses demonstrated his genetic diagnosis was mosaic 45,X[8]/46,X,psu idic(Y)(q12)[32], with the breakpoint in Yq12.ConclusionsOur case proved that it would be beneficial to integrate high-throughput sequencing with cytogenetic technique for precise diagnosis, treatment and genetic counselling.

Efficacy and mechanism of lipoic acid in the treatment of reproductive injury caused by perfluorooctanoic acid.

Environmental pollutants, including perfluorooctanoic acid (PFOA), cause severe reproductive damage to humans and animals. Lipoic acid (LA) is a strong antioxidant that alleviates the oxidative damage caused by heavy metals, environmental toxicants, chemical poisons, etc. Therefore, the present study investigated the curative effect and mechanism of LA in treating spermatogenesis dysfunction caused by PFOA. A rat reproductive injury model was established by gavage of PFOA for consecutive 30 days and then rats were treated with different doses of LA for 42 days. The effects were assessed by ELISA, western blotting, H&E staining and immunofluorescence staining. The results demonstrated that LA had ameliorating effects on PFOA-induced reproductive injury in rats, it increased the sperm counts, and the levels of serum succinate dehydrogenase, lactate dehydrogenase, glutathione peroxidase and superoxide dismutase returned to normal levels following LA treatment. In addition, LA promoted the expression of estradiol, reduced the expression of serum sex hormones, such as follicle-stimulating hormone, androgen receptor (AR) and malondialdehyde in the testes, and restored the structure of seminiferous tubules. Its therapeutic mechanisms include regulating the testicular oxidative stress pathway and hypothalamic-pituitary-testis axis.

Insufficiency of Mrpl40 disrupts testicular structure and semen parameters in a murine model.

Approximately 31% of patients with 22q11.2 deletion syndrome (22q11.2DS) have genitourinary system disorders and 6% of them have undescended testes. Haploinsufficiency of genes on chromosome 22q11.2 might contribute to the risk of 22q11.2DS. In this study, we used mice with single-allele deletion in mitochondrial ribosomal protein L40 (Mrpl40 +/- ) as models to investigate the function of Mrpl40 in testes and spermatozoa development. The penetrance of cryptorchidism in Mrpl40 +/- mice was found to be higher than that in wild-type (WT) counterparts. Although the weight of testes was not significantly different between the WT and Mrpl40 +/- mice, the structure of seminiferous tubules and mitochondrial morphology was altered in the Mrpl40 +/- mice. Moreover, the concentration and motility of spermatozoa were significantly decreased in the Mrpl40 +/- mice. In addition, data-independent acquisition mass spectrometry indicated that the expression of genes associated with male infertility was altered in Mrpl40 +/- testes. Our study demonstrated the important role of Mrpl40 in testicular structure and spermatozoa motility and count. These findings suggest that Mrpl40 is potentially a novel therapeutic target for cryptorchidism and decreased motility and count of spermatozoa.

Glutathione improves testicular spermatogenesis through inhibiting oxidative stress, mitochondrial damage, and apoptosis induced by copper deposition in mice with Wilson disease

Background and objectiveThere are considerable evidence of reproductive impairment in male organisms with Wilson disease (WD). The purpose of this study was to observe spermatogenesis, mitochondrial damage, apoptosis, and the level of oxidative stress in the testes of Wilson disease model TX mice, and to observe the effect and mechanism of glutathione on testicular spermatogenesis. MethodsMice were divided into a normal control group (control group), Wilson disease model TX mice group (WD group), penicillamine-treated TX mice group (penicillamine group) and glutathione-treated TX mice group (glutathione group). Testicular coefficient, histomorphology of testis and epididymis, number of spermatozoa, apoptosis of spermatogenic cells and expression of apoptosis-related proteins were observed. Ultrastructural analysis of mitochondria and mitochondrial membrane potential (MMP) monitored using JC-1 dye were used to detect mitochondrial damage. The levels of malondialdehyde (MDA), glutathione (GSH), catalase (CAT), and reactive oxygen species (ROS) in testicular cells were measured to assess oxidative stress. ResultsTesticular coefficient did not change in mice with Wilson disease. However, the tissue structure of the testicular seminiferous tubules was damaged, and the number of spermatozoa in the epididymal lumen was significantly reduced in WD group. The apoptosis rate in the testes was significantly increased. The protein expression of the pro-apoptotic proteins Bax and Caspase-3 significantly increased, and the expressions of the anti-apoptotic protein Bcl-2 significantly decreased. The levels of ROS and MDA significantly increased, and the levels of CAT and GSH significantly decreased. Mitochondria with abnormal ultrastructure and the rate of JC-1 positive cells were significantly increased in the WD group. After copper chelation by penicillamine, the structure of the testicular seminiferous tubules and the number of spermatozoa in the epididymal lumen were significantly improved. The number of apoptotic cells was significantly reduced. The levels of Bax and Caspase-3 decreased, and the expression of Bcl-2 increased. The contents of CAT and GSH increased, and the levels of ROS and MDA decreased significantly. The abnormal mitochondria and JC-1 positive cells was significantly decreased. The histomorphology of seminiferous tubules, spermatogenic function, apoptosis rate, apoptosis-related proteins, mitochondrial damage, and oxidative stress in Wilson disease TX mice significantly improved after glutathione treatment. ConclusionCopper deposition in Wilson disease can lead to oxidative stress injury, mitochondrial damage, and apoptosis in the testis, leading to the impairment of spermatogenesis. Glutathione may improve testicular spermatogenesis in male Wilson disease TX mice by inhibiting copper deposition-induced oxidative stress, mitochondrial damage, and apoptosis.

Effects of growth hormone on lipid metabolism and sexual development in pubertal obese male rats.

To investigate the effects of growth hormone (GH) on pubertal obese male rats, a rat model of high-fat diet-induced obesity was established in juvenile male rats. The model rats were divided into the treatment group (GH) and the non-treatment group (physiological saline). After 4 weeks, we measured the levels of alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), estrogen (E2), testosterone (T), and insulin-like growth factor (IGF-1). The morphological changes of the liver and testis were assessed, and the expression of aromatase was detected. The levels of ALT, AST, TC, TG, LDL-C, E2, and IGF-1 in the treatment group were significantly lower than in the non-treated model rats (P < 0.001). The levels of HDL-C and T of GH-treated rats were significantly higher than those of the non-treatment group (P < 0.001). Compared with non-treated model rats, GH-treated model rats showed reduced liver steatosis, improved morphological structure of the testicular seminiferous tubules, and an increased number of spermatogenic cells. The treatment group also showed lower expression of aromatase in the liver and testis compared with the non-treatment group. GH partially protected pubertal male rats from obesity-induced lipid metabolic disorder and sexual retardation.

Jujing Zhuyu decoction inhibits apoptosis in rats with asthenozoospermia by regulating the mitochondrial apoptosis pathway.

Jujing Zhuyu decoction (JZD) is a traditional Chinese medicine that effectively improves sperm motility. However, the molecular mechanism of JZD on asthenozoospermia still remains unknown. In this study, we investigated the effect of JZD on the mitochondrial apoptosis pathway in an asthenozoospermia rat model. Sixty Sprague-Dawley rats were randomly divided into five groups-control, tripterygium glycosides (GTW) model, JZD-low (JZD-L), JZD-medium (JZD-M), and JZD-high (JZD-H) groups (n=12/group). GTW was used to generate the asthenozoospermia model. The JZD-L, JZD-M, and JZD-H groups were administered 5, 10, or 15 g kg-1 day-1 of JZD granules respectively, for 4 weeks. Testicular tissue morphology was examined using histological staining, while sperm count was determined using manual and computer-aided semen analyses. Apoptosis of spermatogenic cells was detected with the TUNEL assay, and the expression of proteins and genes related to mitochondrial apoptosis was detected using western blotting and quantitative reverse transcription-polymerase chain reaction respectively. Histomorphological evaluation revealed superior seminiferous tubule structure and arrangement as well as improved spermatogenic cell morphology in the JZD-L, JZD-M, and JZD-H groups compared to those in the model group. Moreover, semen quality and the apoptotic index were significantly improved in the JZD-L, JZD-M, and JZD-H groups compared to those in the model group. Additionally, the mRNA expression and protein abundance of Apaf-1, Bax, Cyto-c, and caspase-3 was reduced, while those of Bcl-2 were increased in all JZD groups compared to those in the model group. JZD reduces the apoptosis rate of sperm cells and significantly promotes sperm survival by regulating the mitochondrial apoptosis pathway. This mechanism provides experimental support for the treatment of asthenozoospermia by JZD.

Absence of MerTK disrupts spermatogenesis in an age-dependent manner

Spermatogenesis is a highly specialized cell differentiation process regulated by the testicular microenvironment. During the process of spermatogenesis, phagocytosis performs an essential role in male germ cell development, and its dysfunction in the testis can cause reproduction defects. MerTK, as a critical protein of phagocytosis, facilitates the removal of apoptotic substrates from the retina and ovaries through cooperation with several phagocytosis receptors. However, its role in mammalian spermatogenesis remains undefined. Here, we found that 30-week-old MerTK−/− male mice developed oligoasthenospermia due to abnormal spermatogenesis. These mice showed damaged seminiferous tubule structure, as well as altered spermatogonia proliferation and differentiation. We also found that Sertoli cells from MerTK−/− mice had decreased phagocytic activity on apoptotic germ cells in vitro. Moreover, a transcriptomic analysis demonstrated that the pivotal genes involved in spermatid differentiation and development changed expression. These results indicate that MerTK is crucial for spermatogenesis, as it regulates the crosstalk between germ cells and Sertoli cells. This provides us insight into the molecular mechanism of MerTK on spermatogenesis and its implications for the diagnosis and treatment of human male infertility.

Agomelatine ameliorates cadmium-induced toxicity through the modification of HMGB-1/TLR-4/NFκB pathway

Cadmium (Cd) has potential hazards on human beings. Consequently, this study was performed to explore the protective effects of agomelatine (AGO), a melatonin receptor agonist, against Cd-induced toxicity in rats. AGO (40 mg/kg/day) was administered orally concomitant with intra peritoneal injection of Cd (0.4 mg/kg/day) for 14 days. Then, blood, biochemical parameters and histological examination of affected organs including, heart and testis, were evaluated. Interestingly, AGO significantly counteracted Cd-induced elevation of serum cardiac enzymes. Similarly, AGO significantly improved the deterioration of serum testosterone level with Cd administration. The oxidative balance was corrected by AGO, as evidenced by decrease malondialdehyde (MDA), and superoxide dismutase activity in cardiac and testicular tissues. Additionally, AGO increased silent information regulator 1 protein (SIRT-1) and decreased High mobility group box 1 (HMGB1), Toll like receptor-4 (TLR-4), and Myd88 levels that subsequently reduced expression of nuclear factor-κB (NF-κB). Moreover, level of apoptotic marker; caspase-3 was inhibited by AGO. In accordance with the biochemical and molecular results, AGO restored structure of cardiac myofibers and seminiferous tubules. Collectively, AGO mitigated cardiac and testicular toxicity of Cd via modulation of SIRT-1/HMGB1 and its downstream pathway.

The effect of aromatase inhibitors against possible testis toxicity in pembrolizumab treated rats.

Pembrolizumab is a monoclonal antibody. Anastrozole is an infertility inhibitor of aromatase. Resveratrol is an antioxidant polyphenol in the reproductive system. This study was planned to demonstrate the protective effects of anastrozole and resveratrol against pembrolizumab-induced reproductive damage. Forty-two Sprague-Dawley rats were used in the study. Groups: The control, Pembrolizumab (PEMB), PEMB + Anastrazol (ANAST), PEMB + Resveratrol (RES), RES, and ANAST groups. At the end of the experiment, rats were euthanased under anaesthesia. Tissue samples were taken from rats for biochemical, histological, and ELISA evaluations. Tissues were subjected to routine tissue follow-up for histological analysis. Biochemically, thiobarbituric acid reactive substance (TBARS), glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) levels were measured. Sperm motility, abnormal sperm rate, and epididymal sperm concentration were examined spermatologically. Serum testosterone and programmed cell death-1 (PD-1) levels were measured using the ELISA. TBARS levels were significantly increased and GSH, SOD, GPx, and CAT levels were mitigated in PEMB-treated rats. Histologically; Control, ANAST, and RES groups testis samples were observed with normal histological appearance. Histological damage was detected in seminiferous tubule structures in testicular tissue in the PEMB group. In treatment groups, this damage was decreased. In addition, PD-1 and testosterone levels were evaluated by the ELISA method. ANAST and RES have therapeutic effects against reproductive damage caused by PEMB.

Investigation of the Effect of Rosmarinic Acid on Cyclophosphamide-Induced Gonadal Toxicity

The aim of this study was to investigate the effect of rosmarinic acid against gonadal toxicity caused by cyclophosphamide, an important anticancer drug. A total of 28 rats were divided into 4 groups, with 7 animals in each group. The groups were created as follows; Group 1 (control) (n = 7): Subjects received only 1 ml of 0.9% saline solution per day intraperitoneally for 14 days. Group 2 (Rosmarinic Acid) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid intraperitoneally for 14 days. Group 3 (Cyclophosphamide) (n = 7): Only 1 ml of 0.9% saline solution was administered intraperitoneally to the subjects for the first 7 days. Cyclophosphamide 20 mg/kg per day was administered intraperitoneally for the last 7 days (from the 8th day). Group 4 (Rosmarinic Acid + Cyclophosphamide) (n = 7): The subjects were given 20 mg/kg Rosmarinic acid daily intraperitoneally for 14 days. From the 8th day of the experiment to the end of the experiment (last 7 days of the experiment), 20 mg/kg cyclophosphamide was given intraperitoneally daily. At the end of the experiment, body weights of all rats were measured first. Afterwards, the weights of testicular tissue samples were measured and the averages of the weights were taken. Routine tissue follows up was performed on the testicular tissues taken. Hematoxylin-eosin staining was applied to tissue sections of 5 μm thickness. As a result of the statistical analysis, it was determined that cyclophosmadine decreased body and testicular weight, but rosmarinic acid had a protective effect on the contrary. It was concluded that cyclophosmadine caused damage to the basement membrane structure, Sertoli, Leydig and germ cells, but these structures were preserved due to the protective effect of rosmarinic acid. Despite the toxic effect of CP, rosmarinic acid is thought to have a significant curative effect on the spermatogenetic process and seminiferous tubule structure in the gonads.

Two distinct Sertoli cell states are regulated via germ cell crosstalk†.

Sertoli cells are a critical component of the testis environment for their role in maintaining seminiferous tubule structure, establishing the blood-testis barrier, and nourishing maturing germ cells in a specialized niche. This study sought to uncover how Sertoli cells are regulated in the testis environment via germ cell crosstalk in the mouse. We found two major clusters of Sertoli cells as defined by their transcriptomes in Stages VII–VIII of the seminiferous epithelium and a cluster for all other stages. Additionally, we examined transcriptomes of germ cell-deficient testes and found that these existed in a state independent of either of the germ cell-sufficient clusters. Altogether, we highlight two main transcriptional states of Sertoli cells in an unperturbed testis environment, and a germ cell-deficient environment does not allow normal Sertoli cell transcriptome cycling and results in a state unique from either of those seen in Sertoli cells from a germ cell-sufficient environment.

Ultrastructural examination of testicular tissues and evaluation of ZO-1 protein levels obtained from azoospermic cases

Spermatogenesis is a process that involves cellular and biochemical processes. The purpose of this study is to understand the role of tight junctions in spermatogenesis by evaluating Zonula occludens-1 (ZO-1) protein levels of cells from the testicular sperm extraction (TESE) biopsies in patients with azoospermia. The study included 40 infertile men with azoospermia. They were divided into two groups, as spermatozoa positive (n=20) and spermatozoa negative (n=20). Testis tissues were examined morphologically, immunohistochemically and ultrastructurally by using transmission electron microscopy (TEM). The structure of seminiferous tubules was deteriorated, degenerated and hyalinized in spermatozoa negative group. ZO-1 expression level was normal in blood-testis barrier of spermatozoa positive group in comparing to lateral surfaces of Sertoli cells spermatozoa negative group. The ultrastructural results were also supported with light microscopy findings. These findings suggest that decrease of the ZO-1 protein expression level in Sertoli cells might be one of the factors involved in the impairment of spermatogenesis in spermatozoa negative group. Therefore, we suggest that tight junctions may have crucial roles in spermatogenesis.

Senescent cells in rabbit, nutria and chinchilla testes—Results from histochemical and immunohistochemical studies

Rabbit, nutria and chinchilla testes were evaluated to compare testicular cellular senescence. There were no major species-specific differences in structure of either seminiferous tubules or interstitial tissue. There, however, were occasional abnormalities in seminiferous tubule structure with there being multinucleated and exfoliated cells present in rabbit testes. Furthermore, there were seminiferous tubules without a lumen that were filled with premeiotic/meiotic cells in nutria; and tubules with vacuolization with there being no post-meiotic cells in chinchillas. There were no differences in distribution or content of acids, total proteins and polysaccharides in the testis of any of the three species. Results using comparative immunohistochemistry procedures indicated the testes contained a few senescent cells in seminiferous tubules with typical morphology and there was a large number of senescent cells in seminiferous tubules of nutrias and chinchillas that had an abnormal structure (P <0.001). Compared to rabbit testes, in which there was the least number of senescent cells in seminiferous tubules, there was a greater abundance of senescence markers in both nutria and chinchilla testes (P < 0.05; P < 0.001, respectively). Furthermore, there were small abundances of caspase 3 and LC3 in the testes of all species. In chinchilla testes, there was a lesser concentration of cholesterol (P < 0.001) and testosterone compared with the other species. Cellular senescence in testes, therefore, can be assessed by detection of morpho-functional disorders of the testis of the three species evaluated in the present study.