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Background: Camels are bred for their milk, meat, wool and hair, transportation, and their excrement as fuel. The seasonal reproduction of camel bull is accompanied by changes in sexual activity, the morphology, and function of the testes. This study aimed to evaluate the seasonal fluctuations in serum testosterone (T) levels as well as total antioxidant capacity (TAC) and malondialdehyde (MDA) levels in the testes of dromedary bulls (Camelus dromedarius) during the rutting and non-rutting seasons. Moreover, the impact of rutting season on the testicular size and histomorphology was also observed. Seventy mature dromedary bulls were divided into a rutting group (n = 35) and a non-rutting group (n = 35). From these bulls, blood samples and testes were collected during the rutting season (October to April) and non-rutting season (May to September) from a local slaughterhouse. Results: All parameters changed significantly during rutting and non-rutting periods in camel bulls. The levels of TAC in testes, and serum T were significantly (P < 0.05) higher in the rutting group than in the non-rutting group. However, testicular MDA was significantly (P < 0.05) lower in the rutting group than in the non-rutting group. TAC was negatively correlated with MDA (r = -0.59, p < 0.01). Moreover, in the rutting group and the non-rutting group, T was positively correlated with levels of TAC (r = 0.66, p < 0.0003). Additionally, testicular size (length, breadth, and thickness) was significantly greater in camels during the rutting season than in camels during the non-rutting season. Moreover, the number and diameter of seminiferous tubules, and spermatogenesis increased during the rutting season, whereas, the collagen content and apoptosis increased during the non-rutting season. Conclusion: This study revealed that the rutting normal breeding season (NBS, rutting group) was associated with higher levels of total antioxidant capacity (TAC), T, and spermatogenic activity while the collagen content, concentrations of MDA (the oxidative stress factor) and apoptosis (an outcome of oxidative stress) were lower than those in the low breeding season (LBS, non-rutting group). In addition, the testicular size and seminiferous tubule diameter and number were higher during the NBS. [ABSTRACT FROM AUTHOR]

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 98.7 Mb and 101.2 Mb on rat chromosome 9. Nucleotide sequence analysis identified 2 bp 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. [ABSTRACT FROM AUTHOR]

Ethanol consumption is increasingly prevalent in communities and has several side effects for humans. Chronic alcohol consumption is often associated with decreased libido and infertility. This study aimed to evaluate the impact of beer on spermatogenesis, proteins, and gene expression of P21 and cyclin D1 in testicular tissue. Twenty-four male mice were assigned to 4 groups. The control group received normal saline, and the experimental groups received alcoholic beer (3 g/kg BW as 20% v/v). After 7, 15, and 35 days, mice were sacrificed, and a part of testicular tissues was stored at −70 °C to measure the protein and gene expression of P21 and cyclin D1 by Western blot and real-time PCR. Moreover, this assay measured the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT). Histomorphometry and histopathology were done. The consumption of alcoholic beer led to increased protein and gene expression of P21 and decreased expression of cyclin D1, as well as reduced levels of SOD and CAT, increased levels of MDA, and subsequent tissue damage in the testicular tissue in a time-dependent manner. Also, the diameter of seminiferous tubules and the thickness of the germinal epithelium were reduced in a time-dependent manner, and the percentage of seminiferous tubules with tubular differentiation, replacement, and negative spermiogenesis coefficients increased significantly. Alcoholic beer disrupted the process of cell division in the testes by reducing the expression of cyclin D1, increasing p21 expression, and inducing oxidative stress, which in turn reduced sperm production and quality. • Alcoholic beer consumption increased protein and gene expression of P21. • Alcoholic beer consumption decreased protein and gene expression of cyclin D1. • Alcoholic beer consumption increased levels of MDA and subsequent tissue damage in the testicular tissue. [ABSTRACT FROM AUTHOR]

The study aimed to assess the effects of water salinity on the sperm parameters, levels of cortisol, LH, FSH, testosterone and antioxidants as well as the testes' histopathology in Barki rams. Fifteen healthy Barki rams (1–1.5 years) were divided into three equal depending on the type of drinking water for nine months. The rams in the tap water group (TW, water that contained 350 ppm of total dissolved salts (TDS). Males in the high saline water group (HSW) were permitted to consume high saline water with 8,934 ppm TDS, whereas those in the second group were permitted to have moderately saline water (MSW, 4,557 ppm TDS). High salt concentration in drinking water had adverse effect on sperm viability, morphology and sperm cell concertation. Nitric oxide and malondialdehyde concentrations in blood were significantly higher in the MSW and HSW groups than in TW. There was a significant decrease in glutathione concentration as well as superoxide dismutase activity in TDS and HSW. Cortisol was most highly concentrated in the HSW, next in the MSW, and least in TW. The testosterone, LH, and FSH concentrations in the HSW and MSW groups were significantly lower than in TW. As the salt concentration in drinking water increases, damage to testicular tissue. The MSW group demonstrating vacuolation of lining epithelial cells with pyknotic nuclei in the epididymis and necrosis and desquamation of spermatogenic cells in seminiferous tubules while HSW group displaying desquamated necrotic cells and giant cell formation in the epididymis, as well as damage to some of the seminiferous tubules and showed congestion, vacuolation of spermatogenic epithelium of seminiferous tubules, and desquamated necrotic spermatogenic epithelium. In conclusion, the salinity of the water has detrimental impacts on the sperm morphology, viability and concentration, hormones and antioxidant levels in Barki rams. [ABSTRACT FROM AUTHOR]

Infertility is an important personal and society disease, of which the male factor represents half of all causes. One of the aspects less studied in male infertility is the immunological testicular microenvironment. Mast cells (MCs), having high potential for regulating spermatogenesis due to fine-tuning the state of the integrative buffer metabolic environment, are one of the most crucial cellular subpopulations of the testicular interstitium. One important component of the MC secretome is proteases that can act as proinflammatory agents and in extracellular matrix (ECM) remodeling. In the testis, MCs are an important cell component of the testicular interstitial tissue (TIT). However, there are still no studies addressing the analysis of a specific MC protease—carboxypeptidase A3 (CPA3)—in cases with altered spermatogenesis. The cytological and histotopographic features of testicular CPA3+ MCs were examined in a study involving 34 men with azoospermia. As revealed, in cases with non-obstructive azoospermia, a higher content of CPA3+ MCs in the TIT and migration to the microvasculature and peritubular tissue of seminiferous tubules were observed when compared with cases with obstructive azoospermia. Additionally, a high frequency of CPA3+ MCs colocalization with fibroblasts, Leydig cells, and elastic fibers was detected in cases with NOA. Thus, CPA3 seems to be of crucial pathogenetic significance in the formation of a profibrogenic background of the tissue microenvironment, which may have direct and indirect effects on spermatogenesis. [ABSTRACT FROM AUTHOR]

Leydig cells are essential components of testicular interstitial tissue and serve as a primary source of androgen in males. A functional deficiency in Leydig cells often causes severe reproductive disorders; however, the transcriptional programs underlying the fate decisions and steroidogenesis of these cells have not been fully defined. In this study, we report that the homeodomain transcription factor PBX1 is a master regulator of Leydig cell differentiation and testosterone production in mice. PBX1 was highly expressed in Leydig cells and peritubular myoid cells in the adult testis. Conditional deletion of Pbx1 in Leydig cells caused spermatogenic defects and complete sterility. Histological examinations revealed that Pbx1 deletion impaired testicular structure and led to disorganization of the seminiferous tubules. Single-cell RNA-seq analysis revealed that loss of Pbx1 function affected the fate decisions of progenitor Leydig cells and altered the transcription of genes associated with testosterone synthesis in the adult testis. Pbx1 directly regulates the transcription of genes that play important roles in steroidogenesis (Prlr, Nr2f2 and Nedd4). Further analysis demonstrated that deletion of Pbx1 leads to a significant decrease in testosterone levels, accompanied by increases in pregnenolone, androstenedione and luteinizing hormone. Collectively, our data revealed that PBX1 is indispensable for maintaining Leydig cell function. These findings provide insights into testicular dysgenesis and the regulation of hormone secretion in Leydig cells. [ABSTRACT FROM AUTHOR]

The blood-testis barrier (BTB) is formed adjacent to the seminiferous basement membrane. It is a distinct ultrastructure, partitioning testicular seminiferous epithelium into apical (adluminal) and basal compartments. It plays a vital role in developing and maturing spermatocytes into spermatozoa via reorganizing its structure. This enables the transportation of preleptotene spermatocytes across the BTB, from basal to adluminal compartments in the seminiferous tubules. Several bioactive peptides and biomolecules secreted by testicular cells regulate the BTB function and support spermatogenesis. These peptides activate various downstream signaling proteins and can also be the target themself, which could improve the diffusion of drugs across the BTB. The gap junction (GJ) and its coexisting junctions at the BTB maintain the immunological barrier integrity and can be the "gateway" during spermatocyte transition. These junctions are the possible route for toxicant entry, causing male reproductive dysfunction. Herein, we summarize the detailed mechanism of all the regulators playing an essential role in the maintenance of the BTB, which will help researchers to understand and find targets for drug delivery inside the testis. [ABSTRACT FROM AUTHOR]

The present research work was carried out on testes procured from six healthy adult non-descript male cats brought for castration/ neutering at Veterinary Clinical Complex of the College in Anand (Gujarat, India). The fibrous connective tissue capsule of testes was made up of outer tunica albugenia and inner tunica vasculosa. The tunica albugenia was thinner as compared to tunica vasculosa layer. The capsule was having collagen fibers and sparce number of elastic fibers. Mean thickness of tunica albuginea was 38.35±1.42 µm. The mediastinal testis was located centrally in the testicular parenchyma. The rete testis was lined by simple cuboidal epithelial cells, presenting prominent oval and horizontally placed nuclei in it. Each lobule of testis was oval to coiled irregular in shape. The mean diameter of seminiferous tubules was 243.37±2.52 µm, whereas the mean height of spermatogenic epithelium was 65.52±0.77 µm. There was average 17-18 Sertoli cells noticed per cross section of seminiferous tubules. The mean height of Sertoli cells was 38.35±1.42 µm. Clusters of Leydig cells were arranged in triangle, rectangle or irregular shape in the interstitial spaces. There was presence of lipid material in the Leydig cells. The mean diameter of Leydig cells was 7.4 µm. The elongated myoid cells were found at the basement membrane of the seminiferous tubules. The tunica albuginea, basement membrane of seminiferous tubules and mediastinal testis were strongly positive for PAS Alcian blue stain indicating more amount of neutral mucopolysaccharides. The seminiferous tubular epithelial cells and interstitial cells were moderately positive for neutral mucopolysaccharides. [ABSTRACT FROM AUTHOR]

Simple Summary: The results of this research revealed significant differences in the tissue morphology and gene expression profiles of Mongolian horses based on age. Specifically, we identified 25 cell clusters and 10 cell types, including spermatogonial and somatic cells, from 17,000 testicular cells of Mongolian horses, constructing a single-cell atlas of the testes. Differential gene expression analysis highlighted distinct patterns in sexually immature and mature testes, with genes related to cellular infrastructure predominating in the former and those associated with spermatogenesis in the latter. Furthermore, we identified marker genes specific to each developmental stage, including APOA1, AMH, TAC3, INHA, SPARC, and SOX9 for the sexually immature stage, and PRM1, PRM2, LOC100051500, PRSS37, HMGB4, and H1-9 for the sexually mature stage. This study aimed to investigate differences in testicular tissue morphology, gene expression, and marker genes between sexually immature (1-year-old) and sexually mature (10-year-old) Mongolian horses. The purposes of our research were to provide insights into the reproductive physiology of male Mongolian horses and to identify potential markers for sexual maturity. The methods we applied included the transcriptomic profiling of testicular cells using single-cell sequencing techniques. Our results revealed significant differences in tissue morphology and gene expression patterns between the two age groups. Specifically, 25 cell clusters and 10 cell types were identified, including spermatogonial and somatic cells. Differential gene expression analysis highlighted distinct patterns related to cellular infrastructure in sexually immature horses and spermatogenesis in sexually mature horses. Marker genes specific to each stage were also identified, including APOA1, AMH, TAC3, INHA, SPARC, and SOX9 for the sexually immature stage, and PRM1, PRM2, LOC100051500, PRSS37, HMGB4, and H1-9 for the sexually mature stage. These findings contribute to a deeper understanding of testicular development and spermatogenesis in Mongolian horses and have potential applications in equine reproductive biology and breeding programs. In conclusion, this study provides valuable insights into the molecular mechanisms underlying sexual maturity in Mongolian horses. [ABSTRACT FROM AUTHOR]

The apelin receptor (APJ) belongs to the member of the G protein‐coupled receptor family, and expression of APJ has been reported in the different cell types of testis. The seminiferous tubules in the testis can be identified as different stages (I–XII). It has been also suggested that different factors could be expressed in stage and cell‐specific manner in the seminiferous tubules. Recently, we also shown that expression of APJ is developmentally regulated in the testis from PND1 to PND42. Therefore, we analyzed the expression of APJ in the testis of adult mice by immunohistochemistry. Immunohistochemistry showed that the APJ was highly specific for the round and elongated spermatids with stage‐dependent changes. The seminiferous tubules at stages I–VII showed APJ immunostaining in the spermatid steps 1–8, not steps of 13–16. The seminiferous tubules at stages IX–XII showed APJ immunostaining in the spermatid steps 9–12. These results suggested the possible role of APJ in the spermiogenesis process. The intratesticular administration of APJ antagonist, ML221 showed a few round spermatids in the seminiferous tubules and some of the tubules with complete absence of round spermatid. Overall, we present evidence that APJ expression in spermatid is dependent on the stages of the seminiferous epithelium cycle and APJ could be involved in the differentiation of round spermatid to elongated spermatid. Research Highlights: The expression of APJ is stage‐ and cell‐specific.The apelin signaling is required for elongation of spermatid and might have a crucial role in spermiogenesis.The intratesticular inhibition of ML221 study suggests that apelin signaling plays important role in spermatid differentiation. [ABSTRACT FROM AUTHOR]

STUDY QUESTION Are there subgroups among patients with cryptozoospermia pointing to distinct etiologies? SUMMARY ANSWER We reveal two distinct subgroups of cryptozoospermic (Crypto) patients based on testicular tissue composition, testicular volume, and FSH levels. WHAT IS KNOWN ALREADY Cryptozoospermic patients present with a sperm concentration below 0.1 million/ml. While the etiology of the severely impaired spermatogenesis remains largely unknown, alterations of the spermatogonial compartment have been reported including a reduction of the reserve stem cells in these patients. STUDY DESIGN, SIZE, DURATION To assess whether there are distinct subgroups among cryptozoospermic patients, we applied the statistical method of cluster analysis. For this, we retrospectively selected 132 cryptozoospermic patients from a clinical database who underwent a testicular biopsy in the frame of fertility treatment at a university hospital. As controls (Control), we selected 160 patients with obstructive azoospermia and full spermatogenesis. All 292 patients underwent routine evaluation for endocrine, semen, and histological parameters (i.e. the percentage of tubules with elongated spermatids). Moreover, outcome of medically assisted reproduction (MAR) was assessed for cryptozoospermic (n = 73) and Control patients (n = 87), respectively. For in-depth immunohistochemical and histomorphometrical analyses, representative tissue samples from cryptozoospermic (n = 27) and Control patients (n = 12) were selected based on cluster analysis results and histological parameters. PARTICIPANTS/MATERIALS, SETTING, METHODS This study included two parts: firstly using clinical parameters of the entire cohort of 292 patients, we performed principal component analysis (PCA) followed by hierarchical clustering on principal components (i.e. considering hormonal values, ejaculate parameters, and histological information). Secondly, for histological analyses seminiferous tubules were categorized according to the most advanced germ cell type present in sections stained with Periodic acid Schif. On the selected cohort of 39 patients (12 Control, 27 cryptozoospermic), we performed immunohistochemistry for spermatogonial markers melanoma-associated antigen 4 (MAGEA4) and piwi like RNA-mediated gene silencing 4 (PIWIL4) followed by quantitative analyses. Moreover, the morphologically defined Adark spermatogonia, which are considered to be the reserve stem cells, were quantified. MAIN RESULTS AND THE ROLE OF CHANCE The PCA and hierarchical clustering revealed three different clusters, one of them containing all Control samples. The main factors driving the sorting of patients to the clusters were the percentage of tubules with elongated spermatids (Cluster 1, all Control patients and two cryptozoospermic patients), the percentage of tubules with spermatocytes (Cluster 2, cryptozoospermic patients), and tubules showing a Sertoli cells only phenotype (Cluster 3, cryptozoospermic patients). Importantly, the percentage of tubules containing elongated spermatids was comparable between Clusters 2 and 3. Additional differences were higher FSH levels (P  < 0.001) and lower testicular volumes (P  < 0.001) in Cluster 3 compared to Cluster 2. In the spermatogonial compartment of both cryptozoospermic Clusters, we found lower numbers of MAGEA4+ and Adark spermatogonia but higher proportions of PIWIL4+ spermatogonia, which were significantly correlated with a lower percentage of tubules containing elongated spermatids. In line with this common alteration, the outcome of MAR was comparable between Controls as well as both cryptozoospermic Clusters. LIMITATIONS, REASONS FOR CAUTION While we have uncovered the existence of subgroups within the cohort of cryptozoospermic patients, comprehensive genetic analyses remain to be performed to unravel potentially distinct etiologies. WIDER IMPLICATIONS OF THE FINDINGS The novel insight that cryptozoospermic patients can be divided into two subgroups will facilitate the strategic search for underlying genetic etiologies. Moreover, the shared alterations of the spermatogonial stem cell compartment between the two cryptozoospermic subgroups could represent a general response mechanism to the reduced output of sperm, which may be associated with a progressive phenotype. This study therefore offers novel approaches towards the understanding of the etiology underlying the reduced sperm formation in cryptozoospermic patients. STUDY FUNDING/COMPETING INTEREST(S) German research foundation CRU 326 (grants to: SDP, NN). Moreover, we thank the Faculty of Medicine of the University of Münster for the financial support of Lena Charlotte Schülke through the MedK-program. We acknowledge support from the Open Access Publication Fund of the University of Münster. The authors have no potential conflicts of interest. TRIAL REGISTRATION NUMBER N/A. [ABSTRACT FROM AUTHOR]

Objective(s): We aimed to investigate the effects of quercetin (QRC) against cyclophosphamide (CP)- induced testicular damage and how it interacts with apoptotic and inflammatory signaling pathways. Materials and Methods: Forty male Wistar rats were randomly divided into four groups, 10 in each group; Control group (corn oil, intragastrically, 14 days), QRC group (100 mg/kg QRC, dissolved in corn oil, 14 days), CP group (200 mg/kg CP, intraperitoneally, single dose on the 7th day), and CP+QRC group (100 mg/kg QRC, intragastrically, 14 days and 200 mg/kg CP, intraperitoneally, single dose on the 7th day). Animals were sacrificed one day after the last QRC application and the effects of quercetin were evaluated by histological, morphometrical, and hormonal parameters. Also, nuclear factor kappa B (NFkB), nuclear factor erythroid 2 related factor 2 (Nrf2), Bcl-2 associated X protein (Bax), and B-cell lymphoma-2 (Bcl-2) immunoreactivities were evaluated immunohistochemically. Results: CP increased the testicular weight/body weight ratio, significantly decreasing body weights and testicular weights. All hormone levels were also reduced significantly. Morphometrically, seminiferous tubules diameter and germinal epithelial thickness decreased, while a significant increase was determined in interstitial field width in addition to histological damage. Furthermore, immunohistochemical findings also indicated that NFkB and Bax immunoreactivity were increased in the CP group, whereas significant decrease was seen in Nrf2 and Bcl-2 immunoreactivity. Apoptotic cell and tubule index were reduced in CP. QRC ensured improvement in all findings. [ABSTRACT FROM AUTHOR]

Snakes represent a wide and diverse group of species and have anatomical particularities, such as the renal sexual segment (RSS), a structure located in the kidneys and formed from the hypertrophy of the urinary ducts and nephrons. This study aims at describing the histological aspects of the RSS of Boa constrictor, Epicrates cenchria and Corallus hortulanus, all of which are Brazilian snake species from the Boidae family. The reproductive system and kidneys of five male specimens of E. cenchria, three male specimens of C. hortulanus and two male specimens of B. constrictor were obtained. Tissue samples were processed histologically and different stains used (Toluidine Blue, Alcian Blue and Periodic Acid Schiff). The histological evaluation of the RSS of E. cenchria, C. hortulanus and B. constrictor shows that the RSS in these species varies when comparing individuals in the reproductive period with those which are not. It also allows for the observation of the segment's secretory activity in animals in the reproductive stage (mature sperm in the lumen of the seminiferous tubules) as well as in those which are not. Finally, the histological evaluation also reveals the variation of the secretion product in individuals in the reproductive period, in those which are not, and also among individuals within the same reproductive stage. [ABSTRACT FROM AUTHOR]

The possible protective effects of Myrtus communis L. (MC) extract on a high fat diet (HFD)-induced testicular injury in a rat model were investigated using histological and biochemical methods. Wistar albino rats were divided into three groups: a standard diet control group; a HFD group; and an HFD+MC group. The HFD and HFD+MC groups were fed with a HFD for 16 weeks. MC extract (100 mg/kg) was given orally five days a week to the rats in the HFD+MC group during the last four weeks of the experiment. Leptin, triglyceride, high-density lipoproteins, cholesterol, estrogen, testosterone, LH and FSH were analyzed in blood serum. Sperm parameters were evaluated from the epididymis. Testicular morphology, proliferative, apoptotic and NADPH oxidase-2 (NOX2)-positive cells were evaluated histologically. Testicular oxidative stress parameters were analyzed biochemically. In the HFD group, lipid and hormone profiles were changed, abnormal spermatozoa, degenerated seminiferous tubules with apoptotic and NOX2-positive cells were increased in number, and sperm motility and germinal proliferative cells decreased compared to the control group. Moreover, testicular malondialdehyde, 8-hydroxy-2-deoxyguanosine and myeloperoxidase levels increased, whereas glutathione and superoxide dismutase levels decreased in the HFD group compared to the control group. All these histological and biochemical features were ameliorated by MC treatment of HFD-fed rats. In conclusion, HFD caused alterations in sperm parameters and testicular morphology by increasing oxidative damage and apoptosis. MC extract may have potential protective effects by inhibiting oxidative damage. [ABSTRACT FROM AUTHOR]

Dynamic nuclear architecture and chromatin organizations are the key features of the mid‐prophase I in mammalian meiosis. The chromatin undergoes major changes, including meiosis‐specific spatiotemporal arrangements and remodeling, the establishment of chromatin loop–axis structure, pairing, and crossing over between homologous chromosomes, any deficiencies in these events may induce genome instability, subsequently leading to failure to produce gametes and infertility. Despite the significance of chromatin structure, little is known about the location of chromatin marks and the necessity of their balance during meiosis prophase I. Here, we show a thorough cytological study of the surface‐spread meiotic chromosomes of mouse spermatocytes for H3K9,14,18,23,27,36, H4K12,16 acetylation, and H3K4,9,27,36 methylation. Active acetylation and methylation marks on H3 and H4, such as H3K9ac, H3K14ac, H3K18ac, H3K36ac, H3K56ac, H4K12ac, H4K16ac, and H3K36me3 exhibited pan‐nuclear localization away from heterochromatin. In comparison, repressive marks like H3K9me3 and H3K27me3 are localized to heterochromatin. Further, taking advantage of the delivery of small‐molecule chemical inhibitors methotrexate (heterochromatin enhancer), heterochromatin inhibitor, anacardic acid (histone acetyltransferase inhibitor), trichostatin A (histone deacetylase inhibitor), IOX1 (JmjC demethylases inhibitor), and AZ505 (methyltransferase inhibitor) in seminiferous tubules through the rete testis route, revealed that alteration in histone modifications enhanced the centromere mislocalization, chromosome breakage, altered meiotic recombination and reduced sperm count. Specifically, IOX1 and AZ505 treatment shows severe meiotic phenotypes, including altering chromosome axis length and chromatin loop size via transcriptional regulation of meiosis‐specific genes. Our findings highlight the importance of balanced chromatin modifications in meiotic prophase I chromosome organization and instability. [ABSTRACT FROM AUTHOR]

Simple Summary: In the testis of infertile patients, we have demonstrated a high prevalence of immune cells (orchitis); their presence together with high FSH levels are associated with alterations in the behavior of nurturing Sertoli cells, which are mostly immature, in the disturbed balance of death and proliferation of germ cells facts leading to reduced sperm production. The experimental model of infertility in rats associated with chronic testicular inflammation, experimental autoimmune orchitis (EAO), shares a common hormonal profile and Sertoli cell and germ cell behavior with orchitis in humans. We support EAO as a useful tool to study how infertility is initiated and progresses, and to search for therapies to improve and/or restore gamete production in infertile patients. Experimental autoimmune orchitis (EAO) is a well-established rodent model of organ-specific autoimmunity associated with infertility in which the testis immunohistopathology has been extensively studied. In contrast, analysis of testis biopsies from infertile patients associated with inflammation has been more limited. In this work, testicular biopsies from patients with idiopathic non-obstructive azoospermia diagnosed with hypospermatogenesis (HypoSp) [mild: n = 9, and severe: n = 11], with obstructive azoospermia and complete Sp (spermatogenesis) (control group, C, n = 9), and from Sertoli cell-only syndrome (SCOS, n = 9) were analyzed for the presence of immune cells, spermatogonia and Sertoli cell (SCs) alterations, and reproductive hormones levels. These parameters were compared with those obtained in rats with EAO. The presence of increased CD45+ cells in the seminiferous tubules (STs) wall and lumen in severe HypoSp is associated with increased numbers of apoptotic meiotic germ cells and decreased populations of undifferentiated and differentiated spermatogonia. The SCs showed an immature profile with the highest expression of AMH in patients with SCOS and severe HypoSp. In SCOS patients, the amount of SCs/ST and Ki67+ SCs/ST increased and correlated with high serum FSH levels and CD45+ cells. In the severe phase of EAO, immune cell infiltration and apoptosis of meiotic germ cells increased and the number of undifferentiated and differentiated spermatogonia was lowest, as previously reported. Here, we found that orchitis leads to reduced sperm number, viability, and motility. SCs were mature (AMH-) but increased in number, with Ki67+ observed in severely damaged STs and associated with the highest levels of FSH and inflammatory cells. Our findings demonstrate that in a scenario where a chronic inflammatory process is underway, FSH levels, immune cell infiltration, and immature phenotypes of SCs are associated with severe changes in spermatogenesis, leading to azoospermia. Furthermore, AMH and Ki67 expression in SCs is a distinctive marker of severe alterations of STs in human orchitis. [ABSTRACT FROM AUTHOR]

Busulfan is an anticancer drug known to cause serious damage to seminiferous tubules in the testes and deplete germ cells in human and animal models. The testicular artery is anastomosed with deferential and cremasteric arteries and is divided into capsular arteries, which give rise to the centripetal arteries and then recurrent arteries. The arterial blood in the testicular tissue is supplied by such a consequent system of arterial vessels, in order from the peripheral to the central area. As anticancer drugs are generally distributed throughout the whole body via the bloodstream and the running and distribution of arteries differ among the testicular areas, we hypothesized that the efficacy of busulfan differs in different testicular areas, particularly between the central and peripheral areas. In this study, busulfan was intraperitoneally injected at 40 mg/kg body weight into C57BL/6J male mice. After 28 days, in busulfan- treated mice, the diameters of seminiferous tubules were significantly higher in the central than in the peripheral area of the testes. The seminiferous tubular areas also significantly decreased in the peripheral areas compared with the central areas. The number of germ cells per seminiferous tubule was significantly higher in the central than in the peripheral area. Sertoli cell nuclei were detached into the lumen in the peripheral area. The number of Leydig cells was significantly lower in the peripheral areas. These data suggest that the effects of busulfan differ between the central and peripheral areas of the testis at 4 weeks after busulfan administration. [ABSTRACT FROM AUTHOR]

The activation of the NLRP3 inflammasome has been implicated in male infertility. Our study aimed to investigate the therapeutic role of Thiolutin (THL), an inhibitor of the NLRP3 inflammasome, on oligoasthenospermia (OA) and to elucidate its mechanisms. Semen from 50 OA and 20 healthy males were analyzed to assess the sperm quality and levels of inflammatory markers. Their correlation was determined using Pearson's correlation coefficient. The BALB/c mice were intraperitoneal injected by cyclophosphamide at 60 mg/kg/day for five days to induce OA, followed by a two-week treatment with THL or L-carnitine. Reproductive organ size and H&E staining were determined to observe the organ and seminiferous tubule morphology. ELISA and western blotting were utilized to measure sex hormone levels, inflammatory markers, and NLRP3 inflammasome levels. Furthermore, male and female mice were co-housed to observe pregnancy success rates. OA patients exhibited a decrease in sperm density and motility compared to healthy individuals, along with elevated levels of IL-1β, IL-18 and NLRP3 inflammasome. In vivo, THL ameliorated OA-induced atrophy of reproductive organs, hormonal imbalance, and improved sperm density, motility, spermatogenesis and pregnancy success rates with negligible adverse effects on weight or liver-kidney function. THL also demonstrated to be able to inhibit the activation of NLRP3 inflammasome and associated proteins in OA mice. THL can improve sperm quality and hormonal balance in OA mice through the inhibition of NLRP3 inflammasome activation. Thus, THL holds promising potential as a therapeutic agent for OA. [ABSTRACT FROM AUTHOR]

Stress granules (SGs) are membraneless ribonucleoprotein (RNP)-based cellular foci formed in response to stress, facilitating cell survival by protecting against damage. Mammalian spermatogenesis should be maintained below body temperature for proper development, indicating its vulnerability to heat stress (HS). In this study, biotin tracer permeability assays showed that the inhibition of heat-induced SG assembly in the testis by 4–8 mg/kg cycloheximide significantly increased the percentage of seminiferous tubules with a damaged blood–testis barrier (BTB). Western blot results additionally revealed that the suppression of heat-induced SG assembly in Sertoli cell line, TM4 cells, by RNA inference of G3bp1/2 aggravated the decline in the BTB-related proteins ZO-1, β-Catenin and Claudin-11, indicating that SGs could protect the BTB against damage caused by HS. The protein components that associate with SGs in Sertoli cells were isolated by sequential centrifugation and immunoprecipitation, and were identified by liquid chromatography with tandem mass spectrometry. Gene Ontology and KEGG pathway enrichment analysis revealed that their corresponding genes were mainly involved in pathways related to proteasomes, nucleotide excision repair, mismatch repair, and DNA replication. Furthermore, a new SG component, the ubiquitin associated protein 2 (UBAP2), was found to translocate to SGs upon HS in TM4 cells by immunofluorescence. Moreover, SG assembly was significantly diminished after UBAP2 knockdown by RNA inference during HS, suggesting the important role of UBAP2 in SG assembly. In addition, UBAP2 knockdown reduced the expression of ZO-1, β-Catenin and Claudin-11, which implied its potential role in the function of the BTB. Overall, our study demonstrated the role of SGs in maintaining BTB functions during HS and identified a new component implicated in SG formation in Sertoli cells. These findings not only offer novel insights into the biological functions of SGs and the molecular mechanism of low fertility in males in summer, but also potentially provide an experimental basis for male fertility therapies. [ABSTRACT FROM AUTHOR]

Purpose: To identify germline mutations related to azoospermia etiology and reproductive potential of surgically retrieved spermatozoa, and to investigate the feasibility of predicting seminiferous tubule function of nonobstructive azoospermic men by transcriptomic profiling of ejaculates. Materials and methods: Sperm specimens were obtained from 30 men (38.4 ± 6 years) undergoing epididymal sperm aspiration for obstructive azoospermia (OA, n = 19) acquired by vasectomy, or testicular biopsy for nonobstructive azoospermia (NOA, n = 11). To evaluate for a correlation with azoospermia etiology, DNAseq was performed on surgically retrieved spermatozoa, and cell-free RNAseq on seminal fluid (n = 23) was performed to predict spermatogenesis in the seminiferous tubule. Results: Overall, surgically retrieved sperm aneuploidy rates were 1.7% and 1.8% among OA and NOA cohorts, respectively. OA men carried housekeeping-related gene mutations, while NOA men displayed mutations on genes involved in crucial spermiogenic functions (AP1S2, AP1G2, APOE). We categorized couples within each cohort according to ICSI clinical outcomes to investigate genetic causes that may affect reproductive potential. All OA-fertile men (n = 9) carried mutations in ZNF749 (sperm production), whereas OA-infertile men (n = 10) harbored mutations in PRB1, which is essential for DNA replication. NOA-fertile men (n = 8) carried mutations in MPIG6B (stem cell lineage differentiation), whereas NOA-infertile individuals (n = 3) harbored mutations in genes involved in spermato/spermio-genesis (ADAM29, SPATA31E1, MAK, POLG, IFT43, ATG9B) and early embryonic development (MBD5, CCAR1, PMEPA1, POLK, REC8, REPIN1, MAPRE3, ARL4C). Transcriptomic assessment of cell-free RNAs in seminal fluid from NOA men allowed the prediction of residual spermatogenic foci. Conclusions: Sperm genome profiling provides invaluable information on azoospermia etiology and identifies gene-related mechanistic links to reproductive performance. Moreover, RNAseq assessment of seminal fluid from NOA men can help predict sperm retrieval during testicular biopsies. [ABSTRACT FROM AUTHOR]

Preclinical research has provided compelling evidence indicating that exposure to hypobaric hypoxia (HH) results in a deterioration of spermatogenesis. This adverse effect extends to the underlying molecular mechanisms, progressively leading to impairments in the seminiferous epithelium and germ cells and alterations in semen parameters. Indeed, several studies have demonstrated that animals exposed to HH, whether in natural high-altitude environments or under simulated hypoxic conditions, exhibit damage to the self-renewal and differentiation of spermatogenesis, an increase in germline cell apoptosis, and structural alterations in the seminiferous tubules. One of the primary mechanisms associated with the inhibition of differentiation and an increase in apoptosis among germ cells is an elevated level of oxidative stress, which has been closely associated with HH exposure. Human studies have shown that individuals exposed to HH, such as mountaineers and alpinists, exhibit decreased sperm count, reduced motility, diminished viability, and increased sperm with abnormal morphology in their semen. This evidence strongly suggests that exposure to HH may be considered a significant risk factor that could elevate the prevalence of male infertility. This literature review aims to provide a comprehensive description and propose potential mechanisms that could elucidate the infertility processes induced by HH. By doing so, it contributes to expanding our understanding of the challenges posed by extreme environments on human physiology, opening new avenues for research in this field. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is a widespread metabolic disease presenting with various complications, including spermatogenic dysfunction. However, the underlying mechanisms are still unclear. Ferroptosis, a novel type of programmed cell death, is associated with much metabolic diseases. Here, we investigated the role of ferroptosis in spermatogenic dysfunction of streptozotocin (STZ)-induced type 1 diabetic mice (diabetic mice), high glucose (HG)-treated GC-2 cells (HG cells) as well as testicular tissues of diabetic patients. We found an accumulation of iron, elevated malondialdehyde level and reduced glutathione level in the testis tissues of diabetic mice and HG cells. Histological examination showed a decrease in spermatogenic cells and spermatids within the seminiferous tubules as well as mitochondrial shrinkage in the testis tissues of diabetic mice. Ferrostatin-1 (Fer-1), the inhibitor of ferroptosis, mitigated ferroptosis-associated iron overload, lipid peroxidation accumulation and spermatogenic dysfunction of diabetic mice. Furthermore, we observed a downregulation of GPX4, FTL and SLC7A11 in diabetic mice and HG cells. Fer-1 treatment and GPX4 overexpression counteracted the effects of HG on cell viability, reactive oxygen species, lipid peroxidation and glutathione via inhibition of ferroptosis. Moreover, we found an elevation of ferroptosis in testicular tissues of diabetic patients. Taken together, our results identify the crucial role of ferroptosis in diabetic spermatogenic dysfunction and ferroptosis may be a promising therapeutic target to improve spermatogenesis in diabetic patients. [Display omitted] • Increased ferroptosis caused spermatogenic dysfunction in type 1 diabetic mice and patients. • Ferrostatin-1 treatment alleviated spermatogenic dysfunction in diabetic mice. • Ferrostatin-1 treatment and GPX4 overexpression attenuated ferroptosis in high glucose treated GC-2 cells. • Inhibition of ferroptosis may be a potential therapy to alleviate diabetic male infertility. [ABSTRACT FROM AUTHOR]

Background: Spermatogenesis is the process of producing mature sperm from Spermatogonial stem cells (SSCs) and this process requires a complex cooperation of different types of somatic and germ cells. Undifferentiated spermatogonia initiate the spermatogenesis and Sertoli cells as the only somatic cells inside of the seminiferous tubule play a key role in providing chemical and physical requirements for normal spermatogenesis, here, we investigated the dysfunction of these cells in non-obstructive azoospermia. Material and method: In this study, we analyzed the expression of sox9 and UTF1 in the non-obstructive human testis by immunohistochemistry. Moreover, we used the KEGG pathway and bioinformatics analysis to reveal the connection between our object genes and protein. Results: The immunohistochemistry analysis of the non-obstructive human seminiferous tubule showed low expression of Sox9 and UTF1 that was detected out of the main location of the responsible cells for these expressions. Our bioinformatics analysis clearly and strongly indicated the relation between UTF1 in undifferentiated spermatogonia and Sox9 in Sertoli cells mediated by POU5F1. Conclusion: Generally, this study showed the negative effect of POU5F1 as a mediator between Sertoli cells as the somatic cells within seminiferous tubules and undifferentiated spermatogonia as the spermatogenesis initiator germ cells in non-obstructive conditions. [ABSTRACT FROM AUTHOR]

The Sertoli cells (SeCs) of the seminiferous tubules secrete a multitude of immunoregulatory and trophic factors to provide immune protection and assist in the orderly development of germ cells. Grafts of naked or encapsulated SeCs have been proved to represent an interesting therapeutic option in a plethora of experimental models of diseases. However, whether SeCs have immunosuppressive or immunomodulatory effects, which is imperative for their clinical translatability, has not been demonstrated. We directly assessed the immunopotential of intraperitoneally grafted microencapsulated porcine SeCs (MC-SeCs) in murine models of fungal infection (Aspergillus fumigatus or Candida albicans) or cancer (Lewis lung carcinoma/LLC or B16 melanoma cells). We found that MC-SeCs (i) provide antifungal resistance with minimum inflammatory pathology through the activation of the tolerogenic aryl hydrocarbon receptor/indoleamine 2,3-dioxygenase pathway; (ii) do not affect tumor growth in vivo; and (iii) reduce the LLC cell metastatic cancer spread associated with restricted Vegfr2 expression in primary tumors. Our results point to the fine immunoregulation of SeCs in the relative absence of overt immunosuppression in both infection and cancer conditions, providing additional support for the potential therapeutic use of SeC grafts in human patients. [ABSTRACT FROM AUTHOR]

As an estrogenic agent, Bisphenol A Dimethacrylate (Bis-DMA) may incite alterations in both the reproductive tract and the neuroendocrine axis, and thus have the potential to affect the proper development, maturity and conceptive performance in animals. We investigated the consequences of 14 weeks of exposure to different concentrations of Bis-DMA on male mouse conceptive performance. Male mice were exposed to Bis-DMA (0, 0.1 mg/L, 1.0 mg/L or 10 mg/L) via drinking water, and the effects on fertility, reproductive organ weights, reproductive hormone levels, sperm counts and testicular histology were assessed. We clearly demonstrate that prolonged exposure of male mice to Bis-DMA negatively affects fertility and reproduction causing significant reductions in sperm counts, non-monotonic effects on serum LH and testosterone levels, increased seminal vesicle weights, lower number of embryonic implantations and viable fetuses, as well as, increased embryonal resorptions in females mated by Bis-DMA treated males. Furthermore, Bis-DMA caused abnormalities in testicular infrastructure with atrophic seminiferous tubules exhibiting intraepithelial vacuolization and disorganization, loss and shedding of germ cells into the lumen, and presence of apoptotic cells. Our data collectively suggest that Bis-DMA adversely affects male fertility and reproduction by interference with normal hormone signaling in the testis, inducing changes in testicular infrastructure and ultimately leading to impaired reproductive function and fertility. [ABSTRACT FROM AUTHOR]

• Chlorophytum borivilianum Santapau & R.R.Fern. (C. borivillianum) used for the treatment of male sexual dysfunction and oligospermia. • Ethanolic extract and saponin enriched n-butanol fraction was evaluated to evolve the underlying biochemical mechanism for spermatogenic potential of C. borivilianum. • Ethanolic extract and saponin enriched n-butanol fraction evaluated to alter fold shift in the mRNA expression of the StAR and 3β-HSD enzymes. • Saponin enriched n-butanol fraction might have interacted with CREB and additionally stimulated LH receptors to amplify the expression of 3β-HSD and StAR mRNA. Traditional applications of the roots of Chlorophytum borivilianum Santapau & R.R.Fern. include spermatogenic potential. Using rats as test subjects, the potential of alcoholic extract (AE-SM) and n-Butanol fraction (NB-SM), which is rich in saponin, to alter the levels of testicular testosterone and cholesterol as well as mRNA expression of the 3β-Hydroxy-∆5- steroid dehydrogenase (3β-HSD) and Acute Steroid Regulatory Protein (StAR) were determined. Histological analyses of testicular tissues were performed, and the quantity of sperm in the seminal fluid was measured. The testosterone concentration increased in AE-SM and NB-SM in a dose-dependent and statistically significant manner. In-vivo mRNA expression of the StAR and 3β-HSD enzymes was also enhanced by the test samples. When compared to control animals, AE-SM and NB-SM increased testicular testosterone by 11.46 and 12.98 fold, respectively, while the extract and fractions revealed a 5.72 and 7.70 fold alteration in StAR mRNA expression. In testicular tissues, AE-SM and NB-SM increased 3β-HSD activity by 4.98 and 9.05 fold, respectively, while changing the expression of the 3β-HSD mRNA by 6.46 and 15.54 factor, respectively. The fluid sampled from the caudal region of the AE-SM and NB-SM indicated an increased sperm concentration, and histological examinations revealed dense Sertoli cell cytoplasm and an increase in the number of sperms in the luminal part of the seminiferous tubules. The components of AE-SM and NB-SM might stimulated testosterone biosynthesis in-vivo by modifying mRNA expression of selected steroidogenic enzymes. By modifying the mRNA expression of the StAR and 3β-HSD, AE-SM and NB-SM can raise the level of testosterone in testicular tissues. As evidenced by an elevated sperm concentration discovered within the seminiferous fluid and histological studies, overexpression of steroidogenic enzymes increased testicular testosterone levels and may have stimulated the spermatogenesis process.Saponin enriched n-butanol fraction might have interacted with CREB and additionally stimulated LH receptors to elevate the StAR and 3β-HSD mRNA expression. [ABSTRACT FROM AUTHOR]

To investigate the effect of age in male quail on testicular weight and histology, sexual libido and semen characteristics, a study was performed on 100 quails at 10, 16, 22, 28 and 34 weeks of age. The body and testicular weights were significantly (p <.05) higher at 16 and 22 than at 28 weeks of age. The circumference and diameter of the seminiferous tubules were significantly (p <.05) higher at 28 and 34 than at 10 and 16 weeks of age. Histological evaluation of testicular slices revealed advanced and effective seminiferous tubes as early as 10 weeks, while spermatogenic activity peaked at 16 weeks of age. The highest semen volume, sperm motility and sperm concentration were observed at weeks of age and then decreased gradually with age. At the same time, the testosterone level and libido were significantly (p <.05) higher at 22 than at 10 weeks of age. Furthermore, the age was positive (p <.05), correlated to seminiferous tubule circumference and diameter, and negative (p <.05), correlated to sperm concentration. In conclusion, the age of quail markedly affected the testicular histological structure, libido, testosterone level and semen characteristics. [ABSTRACT FROM AUTHOR]

Lead is considered a common old chronic toxicant around the world due to expanded environmental pollution, it is likely an inevitable contaminant in food, dairy products, air etc. Also, fipronil is a wide‐ranging effective N‐phenyl pyrazole insecticide which used commonly in agriculture and public health insect control, but until now no adequate data are available on the oxidative stress, cytotoxicity, and mutagenic influence of fipronil and lead or their mixture subchronic exposure. Both xenobiotics (lead and fipronil) exert a harmful impact on reproduction, prompting the exploration of various foods for functional protection. The present study investigated the effects of camel milk treatments on reproductive problems caused by lead acetate with or without mixing with fipronil in male albino rats. Liver oxidative stress, testicular relative weight, sperm analysis, investigation of chromosomal aberration, and histopathological examination of testis were performed. The results showed that the oxidative stress was elevated in rats treated with fipronil, lead acetate, and their mixture, which were reduced through camel milk treatments. Sperm counts were decreased significantly in lead and/or fipronil exposure but significantly elevated with camel milk intoxicated treated. Sperm morphological abnormalities and chromosomal aberrations in intoxicated groups were reduced significantly in camel milk‐treated animals relative to untreated intoxicated groups. Testicular histopathological results showed moderate common degeneration of seminiferous tubules in lead and/or fipronil‐intoxicated rats which were ameliorated by camel milk treatments. Generally, it can be concluded that lead and fipronil together in a mixture resulted in or induced severe reproductive problems and oxidative stress over lead or fipronil alone. Camel milk treatment significantly decreased the harmful oxidative stress in reproductive as well as the mutagenicity disorder associated with lead and fipronil exposure in male albino rats. [ABSTRACT FROM AUTHOR]

Sofosbuvir treatment regimens for chronic HCV infection have recently been linked to extra hepatic side effects. This study aimed to show how sofosbuvir affected the adult male albino rat testis. Forty adult male albino rats were used. The rats were equally split into two main groups (I and II), then each group subdivided into two subgroups (A and B). Each rat in group I (control) received 0.5 ml of distilled water every day for four weeks. Each rat in group II (sofosbuvir-treated) received 0.5 ml of distilled water containing 7.2 mg of sofosbuvir every day for four weeks. After four weeks (subgroups IA and IIA) and eight weeks (subgroups IB and IIB) of treatment, the rats were sacrificed. Histological, biochemical, and morphometric studies on the testes were conducted. The data were analyzed. Examination of testes of sovaldi treated group revealed histopathological changes. Biochemical and morphometric analysis showed reduced levels of reduced glutathione and seminiferous tubule epithelial height respectively. Following a 4-week drug withdrawal period, the testes only partially recovered. We concluded that sofosbuvir induced deteriorating changes in the adult male albino rats' testes. These changes were proved by histological and biochemical studies. These changes were incompletely reversible after cession of treatment. Researches investigating the effect of adding drugs that have antioxidant properties during sofosbuvir therapy are recommended. [ABSTRACT FROM AUTHOR]

Deltamethrin is a widely used synthetic pyrethroid pesticide. It causes reproductive toxicity. Aim of the work: it evaluates the impact of vitamin E in restoration of the testicular integrity of albino rats after toxicity induced by Deltamethrin. Thirty-six adult male albino rats were included, and they were further sub-divided into four experimental groups; Group A: six rats served as controls. Group B (Model): 10 rats equally divided into two sub-groups (B1): the rats received deltamethrin dissolved in oil in a dose of 0.6 mg/kg/daily by nasogastric gavage for 2 weeks. (B2): the rats received Deltamethrin in the same dose of group B1 for 1 month. Group C (Protected): 10 rats equally divided into two sub-groups (C1): the rats received deltamethrin orally 0.6 mg/kg/day concomitant with Vitamin E dissolved in 1 ml of corn oil in a dose 200 mg/kg/day by nasogastric gavage for 2 weeks. (C2): the rats received deltamethrin concomitant with Vitamin E in the same dose of group C1 for 1 month. Group D (Treatment): 10 rats received deltamethrin for 1 month followed by Vitamin E for another month in the same previously prescribed doses. Significant decreases in serum testosterone level, GSH, catalase activity, and significant increase in MDA in the deltamethrin-treated group were detected. Moreover, histological and ultrastructural examinations of the testis seminiferous tubules showed detrimental alterations in the deltamethrin group which were duration dependent. Vitamin E administration reversed such alterations. Vitamin E ameliorates the testicular dysfunction caused by Deltamethrin. [ABSTRACT FROM AUTHOR]

Despite numerous female contraceptive options, nearly half of all pregnancies are unintended. Family planning choices for men are currently limited to unreliable condoms and invasive vasectomies with questionable reversibility. Here, we report the development of an oral contraceptive approach based on transcriptional disruption of cyclical gene expression patterns during spermatogenesis. Spermatogenesis involves a continuous series of self-renewal and differentiation programs of spermatogonial stem cells (SSCs) that is regulated by retinoic acid (RA)-dependent activation of receptors (RARs), which control target gene expression through association with corepressor proteins. We have found that the interaction between RAR and the corepressor silencing mediator of retinoid and thyroid hormone receptors (SMRT) is essential for spermatogenesis. In a genetically engineered mouse model that negates SMRT-RAR binding (SMRTmRID mice), the synchronized, cyclic expression of RAR-dependent genes along the seminiferous tubules is disrupted. Notably, the presence of an RA-resistant SSC population that survives RAR de-repression suggests that the infertility attributed to the loss of SMRT-mediated repression is reversible. Supporting this notion, we show that inhibiting the action of the SMRT complex with chronic, low-dose oral administration of a histone deacetylase inhibitor reversibly blocks spermatogenesis and fertility without affecting libido. This demonstration validates pharmacologic targeting of the SMRT repressor complex for non-hormonal male contraception. [ABSTRACT FROM AUTHOR]

Background: PRAME constitutes one of the largest multi-copy gene families in Eutherians, encoding cancer-testis antigens (CTAs) with leucine-rich repeats (LRR) domains, highly expressed in cancer cells and gametogenic germ cells. This study aims to elucidate genetic interactions between two members, Pramex1 and Pramel1, in the mouse Prame family during gametogenesis using a gene knockout approach. Result: Single-gene knockout (sKO) of either Pramex1 or Pramel1 resulted in approximately 7% of abnormal seminiferous tubules, characterized by a Sertoli-cell only (SCO) phenotype, impacting sperm count and fecundity significantly. Remarkably, sKO female mice displayed normal reproductive functions. In contrast, Pramex1/Pramel1 double knockout (dKO) mice exhibited reduced fecundity in both sexes. In dKO females, ovarian primary follicle count decreased by 50% compared to sKO and WT mice, correlating with a 50% fecundity decrease. This suggested compensatory roles during oogenesis in Pramex1 or Pramel1 sKO females. Conversely, dKO males showed an 18% frequency of SCO tubules, increased apoptotic germ cells, and decreased undifferentiated spermatogonia compared to sKO and WT testes. Western blot analysis with PRAMEX1- or PRAMEL1-specific antibodies on sKO testes revealed compensatory upregulation of each protein (30–50%) in response to the other gene's deletion. Double KO males exhibited more severe defects in sperm count and litter size, surpassing Pramex1 and Pramel1 sKO accumulative effects, indicating a synergistic enhancement interaction during spermatogenesis. Additional experiments administering trans-retinoic acid (RA) and its inhibitor (WIN18,446) in sKO, dKO, and WT mice suggested that PRAMEX1 and PRAMEL1 synergistically repress the RA signaling pathway during spermatogenesis. Conclusion: Data from sKO and dKO mice unveil a synergistic interaction via the RA signaling pathway between Pramex1 and Pramel1 genes during gametogenesis. This discovery sets the stage for investigating interactions among other members within the Prame family, advancing our understanding of multi-copy gene families involved in germ cell formation and function. [ABSTRACT FROM AUTHOR]

Objective. In the testicular tissue stroma, matrix proteins such as laminin and fibronectin play a critical role in transmitting signals under normal and stressful conditions to Leydig cells. Heat stress is considered one of the stressful conditions in the testicular tissue. This study has investigated the protective effects of chrysin on the distribution of laminin and fibronectin around Leydig cells under heat stress. Methods. Rats were divided into four groups, including sham (A), heat stress (B), heat stress with chrysin treatment (50 mg/kg) (C), and heat stress with chrysin treatment (75 mg/kg) (D). At the end of the treatment period, on the 21st day, testosterone serum levels were evaluated, and the testes were also assessed to determine the changes in gene expression and distribution of laminin, fibronectin, and caspase-3 proteins. Results. Exposure to heat stress in Group B resulted in increased levels of caspase-3, decreased activity, height, and diameter of seminiferous tubules, and altered distribution of laminin and fibronectin proteins around the Leydig cells when compared to the sham group (p < 0.05). Treatment with chrysin 75 resulted in a significant decrease in caspase-3 levels, improved testosterone levels, and increased distribution of laminin and fibronectin around Leydig cells compared to group B (p < 0.05). In addition, the group treated with chrysin 75 showed improved seminiferous histological activity markers in comparison to group B (p < 0.05). Conclusion. Following heat stress, concurrent with disrupted steroidogenesis, an evident change happens in the distribution pattern of the extracellular matrix (ECM) proteins of the testis, which may indicate the transmission of stress signals from the ECM to the Leydig cells. Treatment of chrysin in 75 mg/kg dose can reduce the damage caused by heat stress on the ECM and Leydig cells. [ABSTRACT FROM AUTHOR]

In recent years, adipose-derived stem cells (ADSCs) and derived exosomes (ADSC-Ex) have been investigated for their therapeutic potential in various diseases due to their satisfactory differentiation and regeneration ability. We aimed to explore the potential treatment of ADSCs and ADSC-Ex for testicular injury caused by cisplatin. ADSCs and ADSC-Ex s were identified and extracted to treat the rat model with testicular injury caused by cisplatin. Then the immunohistochemistry and Enzyme linked immunosorbent assay (ELISA) were used to detect the potential treatment of ADSCs and ADSC-Ex. We found that ADSCs and ADSC-Ex significantly improved the testicular tissue damage, increased the number of germ cells, and improved the arrangement of the seminiferous tubules. The levels of malondialdehyde and testosterone were also improved. We speculated that ADSCs and ADSC-Ex may alleviate the testicular injury caused by cisplatin. [ABSTRACT FROM AUTHOR]

Background: An increase in the temperature of the testis is associated with damage to the epithelium of seminiferous tubules and disruption of sperm production. Objective: The current study aimed to investigate the effect of the Sertoli cell-conditioned medium (SCCM) on the blood-testis-barrier associated genes and spermatogenesis process following scrotal hyperthermia. Materials and Methods: In this experimental study, 40 adult NMRI mice (8 wk, 25-30 gr) were allocated into 4 groups: I) control, II) DMEM (10 µl Dulbecco's Modified Eagle Medium), III) scrotal hyperthermia, and IV) scrotal hyperthermia+SCCM (10 µl SCCM). Hyperthermia was induced by placing the mice scrotum in water at 43°C for 20 min every other day for 10 days. Mice were treated every other day for 5 wk. Then the animals were euthanized, and the tails of epididymis were removed to analyze sperm parameters, testis were taken for stereological assessment, reactive oxygen spices and glutathione levels, and the expression of Ocln, Gja1, Cdh2, and Itgb1. Results: The results of sperm analysis indicated that SCCM-treated mice significantly increased sperm count and motility and reduced DNA fragmentation. In addition, histological and molecular findings showed that the volume of testicular tissue, the number of germ cells, the glutathione level, and the expression of Ocln, Gja1, Cdh2, and Itgb1 genes were significantly increased in the SCCM-treated mice. Conclusion: Findings suggest that growth factors of SCCM stimulate the proliferation and differentiation of germ cells through paracrine effects and upregulate the blood-testis-barrier-associated genes in mice subjected to scrotal hyperthermia. [ABSTRACT FROM AUTHOR]

The branched-chain amino acids (BCAAs: leucine, isoleucine and valine) are essential for animal growth and metabolic health. However, the effect of valine on male reproduction and its underlying molecular mechanism remain largely unknown. Here, we showed that l -valine supplementation (0.30% or 0.45%, water drinking for 3 weeks) did not change body and testis weights, but significantly altered morphology of sertoli cells and germ cells within seminiferous tubule, and enlarged the space between seminiferous tubules within mouse testis. l -valine treatment (0.45%) increased significantly the Caspase3/9 mRNA levels and CASPASE9 protein levels, therefore induced apoptosis of mouse testis. Moreover, gene expression levels related to autophagy (Atg5 and Lamb3), DNA 5 mC methylation (Dnmt1, Dnmt3a, Tet2 and Tet3), RNA m6A methylation (Mettl14, Alkbh5 and Fto), and m6A methylation binding proteins (Ythdf1/2/3 and Igf2bp1/2) were significantly reduced. Protein abundances of ALKBH5, FTO and YTHDF3 were also significantly reduced, but not for ATG5 and TET2. Testis transcriptome sequencing detected 537 differentially expressed genes (DEGs, 26 up-regulated and 511 down-regulated), involved in multiple important signaling pathways. RT-qPCR validated 8 of 9 DEGs (Cd36, Scd1, Insl3, Anxa5, Lcn2, Hsd17b3, Cyp11a1, Cyp17a1 and Agt) to be decreased significantly, consistent with RNA-seq results. Taken together, l -valine treatment could disturb multiple signaling pathways (autophagy and RNA methylation etc.), and induce apoptosis to destroy the tissue structure of mouse testis. • l -valine intake changed cell morphology and tissue structure of mouse testis in a dose-dependent manner. • l -valine intake induced apoptosis via the Caspase pathway. • l -valine affected autophagy and nucleic acid methylation pathways. • RNA-seq identified 537 DEGs altered by l -valine, most of which down-regulated and involved in multiple signaling pathways. [ABSTRACT FROM AUTHOR]

Background: The association between thiopurine use and testicular reproductive functions remains unclear. In this study, we investigated whether thiopurines affect testicular functions based on the NUDT15 genotypes using Nudt15R138C knock-in mice. Methods: The male Nudt15R138C knock-in mice (9–12 weeks) were treated with mercaptopurine (MP: 0.5 mg/kg/day) for 4 or 12 weeks. To examine reversibility, some mice were maintained for a further 12 weeks under MP-free condition. Results: After MP treatment for 4 weeks, Nudt15R138C/R138C mice exhibited a significant reduction of testis weight compared to Nudt15+/+ mice and Nudt15+/R138C mice. The epithelial height and diameter of seminiferous tubules were significantly reduced in Nudt15R138C/R138C mice compared to Nudt15+/+ and Nudt15+/R138C mice. Apoptotic cells were significantly increased in Nudt15R138C/R138C mice, and most of apoptotic cells were spermatogonia. There were no significant changes in sperm counts and sperm morphology in MP-treated Nudt15R138C/R138C mice after 4-week MP treatment. On the other hand, after MP treatment for 12 weeks, the Nudt15+/R138C mice, but not Nudt15+/+ mice, exhibited a significant reduction in the testis weight and atrophic changes of seminiferous tubules, but these changes disappeared after 12-week rearing under MP-free condition. Despite a significant increase in abnormal sperm rate, there were no changes in the ability to conceive. No differences in serum levels of follicle-stimulating hormone or testosterone were observed between MP-treated Nudt15+/R138C and Nudt15+/+ mice after 12-week MP treatment. Conclusions: Thiopurines exert harmful effects on testicular reproductive function according to host NUDT15 genotypes. [ABSTRACT FROM AUTHOR]

Patients with Idiopathic non-obstructive azoospermia (iNOA) can achieve fertility by extracting testicular sperm through microdissection testicular sperm extraction (mTESE). But more than half of iNOA patients still cannot benefit from mTESE. In recent years, some studies had reported that serum hormones may be related to the outcome of sperm retrieval, but few had been verified. We hope to obtain a predictive method that is convenient for clinical application and can help judge the outcome of sperm extraction before implementing mTESE. We performed a retrospective analysis of NOA patients who underwent mTESE in the same andrology center from June 2020 to November 2022. A total of 261 patients with complete data were collected, logistic regression analysis was performed and a predictive model was constructed. Then, from December 2022 to May 2023, one prospective cohort of 48 NOA patients who met the inclusion criteria from the same center was recruited to validate the risk prediction model. We successfully constructed a logistic regression model to predict the outcome of iNOA patients undergoing mTESE and found that higher serum anti-Müllerian hormone (AMH) levels were associated with failure sperm retrieval, resulting in an AMH cut-off of 2.60 ng/ml. The area under the receiver operating curve was 0.811, the sensitivity was 0.870, and the specificity was 0.705. Decision curve analysis demonstrated that the threshold probability was above 4%, and unnecessary mTESE could be reduced using this model. In a prospective cohort at the same center, 85.42% (41/48) of iNOA patients correctly identified the mTESE outcome using this model. A logistic regression model with AMH as an independent predictor can predict mTESE outcomes in iNOA patients. Preoperative selection of mTESE in patients with iNOA using this model had clinical benefit in reducing unnecessary surgery. The model demonstrated good accuracy in a small prospective cohort validation. [ABSTRACT FROM AUTHOR]

Background: Phytoestrogens are non-estradiol molecules categorized as endocrine-disrupting chemicals, which means they can alter the hormonal equilibrium and cause reproductive problems. Studies suggested Hibiscus sabdariffa (H. sabdariffa) possible use as alternative to hormone replacement therapy in ovarian hypofunction patients. Salvia officinalis (S. officinalis) contains phytoestrogen steroids and isoflavonoids. The purpose of this research was to examine the estrogenic effects of H. sabdariffa and S. officinalis aqueous extracts on male reproductive hormones and testicular histology and function. Methods: Three groups of six male rats each were utilized as follows: control (distilled water), H. sabdariffa (500 mg/kg aqueous extract), and S. officinalis (500 mg/kg aqueous extract). Blood, epididymis. Testis and serum samples were gathered after 30 days for biochemical analysis, sperm characteristics, and histological evaluations. Results: Compared to the control and S. officinalis groups, H. sabdariffa caused a significant reduction in body weight (BW) and percent BW gain (%BWG). Relative to the control rats, the two extracts dramatically reduced testicular weight, sperm motility, concentration, and viability while raising abnormalities. Furthermore, the herbal extract groups demonstrated a significant decrease in blood testosterone levels while increasing serum prolactin, FSH, and LH levels. Consumption of both herbs also caused testicular histological changes such as degenerated seminiferous tubules, with a diminished number of mature spermatozoa in the tubular lumen, reduced diameter of seminiferous tubules, and the presence of exfoliated cells in the tubular lumen. Conclusion: The results of this study revealed that the aqueous extracts of H. sabdariffa and S. officinalis may negatively impact male fertility. However, more research is needed to validate these findings and investigate the mechanistic components of these impacts. [ABSTRACT FROM AUTHOR]

Background and Purpose: Damage to the testis following exposure to ionizing radiation has become an urgent problem to be solved. Here we have investigated if inhibition of p38 mitogen‐activated protein kinase (p38MAPK) signalling could alleviate radiation‐induced testicular damage. Experimental Approach: In mice exposed to whole body radiation (2‐6 Gy), morphological changes of the epididymis and testis was measured by histochemical staining. immunohistochemical and immunofluorescence procedures and western blotting were used to monitor expression and cellular location of proteins. Expression of genes was assessed by qPCR and RNA‐Seq was used to profile gene expression. Key Results: Exposure to ionizing radiation induced dose‐dependent damage to mouse testis. The sperm quality decreased at 6 and 8 weeks after 6 Gy X‐ray radiation. Radiation decreased PLZF+ cells and increased SOX9+ cells, and affected the expression of 969 genes, compared with data from non‐irradiated mice. Expression of genes related to p38MAPK were enriched by GO analysis and were increased in the irradiated testis, and confirmed by qPCR. Levels of phospho‐p38MAPK protein increased at 28 days after irradiation. In irradiated mice, SB203580 treatment increased spermatozoa, SOX9+ cells, the area and diameter of seminiferous tubules, sperm movement rate and density. Furthermore, SB203580 treatment increased SCP3+ cells, accelerating the process of spermatogenesis. Conclusion and Implications: Exposure to ionizing radiation clearly changed gene expression in mouse testis, involving activation of p38MAPK signalling pathways. Inhibition of p38MAPK by SB203580 partly alleviated the testicular damage caused by radiation and accelerated the recovery of sperms through promoting spermatogenesis. [ABSTRACT FROM AUTHOR]

In mammals, females undergo reproductive cessation with age, whereas male fertility gradually declines but persists almost throughout life. However, the detailed effects of ageing on germ cells during and after spermatogenesis, in the testis and epididymis, respectively, remain unclear. Here we comprehensively examined the in vivo male fertility and the overall organization of the testis and epididymis with age, focusing on spermatogenesis, and sperm function and fertility, in mice. We first found that in vivo male fertility decreased with age, which is independent of mating behaviors and testosterone levels. Second, overall sperm production in aged testes was decreased; about 20% of seminiferous tubules showed abnormalities such as germ cell depletion, sperm release failure, and perturbed germ cell associations, and the remaining 80% of tubules contained lower number of germ cells because of decreased proliferation of spermatogonia. Further, the spermatozoa in aged epididymides exhibited decreased total cell numbers, abnormal morphology/structure, decreased motility, and DNA damage, resulting in low fertilizing and developmental rates. We conclude that these multiple ageing effects on germ cells lead to decreased in vivo male fertility. Our present findings are useful to better understand the basic mechanism behind the ageing effect on male fertility in mammals including humans. Declined fertility in aged male mice are associated with decreased sperm production in aged testes and functional sperm defects in aged epididymides, which are independent of testosterone levels. [ABSTRACT FROM AUTHOR]

Purpose: Varicocele is a pathological condition that affects approximately 20% of adult men, causing infertility and sperm deterioration. The aim of our study was to investigate the efficacy of vitamin D (Vit. D) in the pathophysiology of varicocele. We performed biochemical, gene expression analyses and histopathological to evaluate the efficacy of vitamin D in the experimental varicocele model. Materials and Methods: In the study, 30 adult male Wistar Albino rats were used. The rats were divided into 3 groups equally as control group, experimental group (varicocele), treatment group (varicocele + D vit.). The treatment group received 500 IU/kg D vit. intramuscularly. Results: Histopathological, TRPM2-8 gene expression and biochemical analyses were performed on testicular and blood samples collected at the end of the experiment. The experimental group showed a deterioration in tubular structure, a decrease in total antioxidant levels and an increase in total oxidant levels. The treatment group, on the other hand, showed an increase in TAS, a decrease in TOS and a beneficial improvement in tubular structure disorders. Analysis of gene expression levels showed that TRPM2-8 expression levels were significantly increased in the varicocele group and decreased in the treatment group. Conclusion: In the varicocele model, the use of vitamin D had a significant effect on TRPM2-8 gene level, pathological seminiferous tubules and biochemical values. Further studies are needed to determine the clinical application of vitamin D in varicocele disease. [ABSTRACT FROM AUTHOR]

Background: Sodium nitrite (NaNO2) is a chemical substance used to enhance taste, add color, and keep food products fit for consumption for a longer time. NaNO2 gives rise to a negative adverse effect on male reproductive function. Odontonema cuspidatum (OC) is a natural plant that possesses antioxidant capacity. Aim: Our research evaluates the potential beneficial effect of OC extract on the harmful effects caused by NaNO2 on the testicular tissue and sperm characteristics of male rats. Methods: Four groups with a total of forty rats: the control, the NaNO2 -received group, the OC-administered group, and the fourth group received both NaNO2 and OC. All groups were administered daily for two months. Sperm characteristics, testicular antioxidant status, qRT-PCR, and histopathological changes were evaluated. Results: Coadministration of NaNO2 and OC, in comparison with NaNO2 alone, contributed to a notable enhancement in acrosomal integrity, decreasing sperm abnormalities and restoring serum testosterone levels. Moreover, such coadministration reduced the oxidative stress marker, malondialdehyde (MDA), and increased superoxide dismutase (SOD) in testicular tissue, lowering TNF-α gene expression, and increasing the expression of P450scc and StAR genes. In addition, the NaNO2 and OC combination decreased the testicular histopathological changes and the Caspase-3 and Proliferating cell nuclear antigen (PCNA) immunoexpression in seminiferous tubules compared with the NaNO2 group. Conclusion: The extract of OC exhibited the ability to decrease oxidative stress and ameliorate the detrimental effects caused by NaNO2. [ABSTRACT FROM AUTHOR]

Non‐obstructive azoospermia (NOA) is the most severe form of human male infertility, and the genetic causes of NOA with meiotic arrest remain largely unclear. In this study, we identified novel compound heterozygous MEIOB variants (c.814C > T: p.R272X and c.976G > A: p.A326T) and a previously undescribed homozygous non‐canonical splicing variant of MEIOB (c.528 + 3A > C) in two NOA‐affected individuals from two irrelevant Chinese families. MEIOB missense variant (p.A326T) significantly reduced protein abundance and nonsense variant (p.R272X) produced a truncated protein. Both of two variants impaired the MEIOB‐SPATA22 interaction. The MEIOB non‐canonical splicing variant resulted in whole Exon 6 skipping by minigene assay, which was predicted to produce a frameshift truncated protein (p.S111Rfs*32). Histological and immunostaining analysis indicated that both patients exhibited a similar phenotype as we previously reported in Meiob mutant mice, that is, absence of spermatids in seminiferous tubules and meiotic arrest. Our study identified three novel pathogenic variants of MEIOB in NOA patients, extending the mutation spectrum of the MEIOB and highlighting the contribution of meiotic recombination related genes in human fertility. [ABSTRACT FROM AUTHOR]

The indirect assessment of adverse effects on fertility in cynomolgus monkeys requires that tissue sections of the testis be microscopically evaluated with awareness of the stage of spermatogenesis that a particular cross-section of a seminiferous tubule is in. This difficult and subjective task could very much benefit from automation. Using digital whole slide images (WSIs) from tissue sections of testis, we have developed a deep learning model that can annotate the stage of each tubule with high sensitivity, precision, and accuracy. The model was validated on six WSI using a six-stage spermatogenic classification system. Whole slide images contained an average number of 4938 seminiferous tubule cross-sections. On average, 78% of these tubules were staged with 29% in stage I-IV, 12% in stage V-VI, 4% in stage VII, 19% in stage VIII-IX, 18% in stage X-XI, and 17% in stage XII. The deep learning model supports pathologists in conducting a stage-aware evaluation of the testis. It also allows derivation of a stage-frequency map. The diagnostic value of this stage-frequency map is still unclear, as further data on its variability and relevance need to be generated for testes with spermatogenic disturbances. [ABSTRACT FROM AUTHOR]

Royal jelly (RJ) was applied as a supplement in feed to examine its impact on rabbit buck fertility. The aims were to evaluate the effect of RJ on sperm progressive motility and on histology of testis in rabbits. Twenty male rabbits were divided into four groups, each consisting of five bucks: Control (0mg), T1 {100mg RJ/kg body weight (BW)}, T2 (200mg RJ/kg BW), and T3 (300mg RJ/kg BW), with dietary treatment administered for 8 weeks. Sperm progressive motility (%), testis weight (g) and diameter of seminiferous tubule (µm) were measured. T1 group showed significantly higher (P<0.05) sperm progressive motility and seminiferous tubule diameter compared to the other groups, while an excessive concentration of RJ had a negative impact on the sperm quality, potentially because its components may result in an imbalance in redox reactions due to antioxidative activity. It is concluded that an appropriate dose of RJ improved the reproductive characteristics, but the results do depend on the dose administered to the rabbit. [ABSTRACT FROM AUTHOR]

Background: An increase in the temperature of the testis is associated with damage to the epithelium of seminiferous tubules and disruption of sperm production. Objective: The current study aimed to investigate the effect of the Sertoli cell-conditioned medium (SCCM) on the blood-testis-barrier associated genes and spermatogenesis process following scrotal hyperthermia. Materials and Methods: In this experimental study, 40 adult NMRI mice (8 wk, 25-30 gr) were allocated into 4 groups: I) control, II) DMEM (10 μl Dulbecco’s Modified Eagle Medium), III) scrotal hyperthermia, and IV) scrotal hyperthermia+SCCM (10 μl SCCM). Hyperthermia was induced by placing the mice scrotum in water at 43°C for 20 min every other day for 10 days. Mice were treated every other day for 5 wk. Then the animals were euthanized, and the tails of epididymis were removed to analyze sperm parameters, testis were taken for stereological assessment, reactive oxygen spices and glutathione levels, and the expression of Ocln, Gja1, Cdh2, and Itgb1. Results: The results of sperm analysis indicated that SCCM-treated mice significantly increased sperm count and motility and reduced DNA fragmentation. In addition, histological and molecular findings showed that the volume of testicular tissue, the number of germ cells, the glutathione level, and the expression of Ocln, Gja1, Cdh2, and Itgb1 genes were significantly increased in the SCCM-treated mice. Conclusion: Findings suggest that growth factors of SCCM stimulate the proliferation and differentiation of germ cells through paracrine effects and upregulate the blood-testis-barrier-associated genes in mice subjected to scrotal hyperthermia. [ABSTRACT FROM AUTHOR]

Erhualian pigs exhibit one of the highest reproductive rates globally, and cryptorchidism is a crucial factor affecting reproductive abilities of boars. This investigation focused on cryptorchid tissues from Erhualian pigs, where the histological structure of cryptorchidism was observed using specialized staining. In addition, protein expression of P53/NF-κB in cryptorchid tissues was assessed using Western blot and immunohistochemistry. In comparison to normal Erhualian testes, Masson's trichrome staining indicated a reduction in collagen fibers in the connective tissue and around the basal membrane of the seminiferous tubules in cryptorchid testes. Moreover, collagen fiber distribution was observed to be disordered. Verhoeff Van Gieson (EVG) and argyrophilic staining demonstrated brownish-black granular nucleoli organized regions in mesenchymal cells and germ cells. When compared to normal testicles, the convoluted seminiferous tubules of cryptorchids exhibited a significantly reduced number and diameter (p < 0.01). Notably, VEGF/EGFR and P53/NF-κB expression in cryptorchidism significantly differed from that in normal testes. In particular, the expression of VEGF and P53 in cryptorchid tissues was significantly higher than that in normal testes tissues, whereas the expression of EGFR in cryptorchid tissues was significantly lower than that in normal testes tissues (all p < 0.01). NF-κB expressed no difference in both conditions. The expressions of VEGF and NF-κB were observed in the cytoplasm of testicular Leydig cells and spermatogenic cells, but they were weak in the nucleus. EGFR and P53 were more positively expressed in the cytoplasm of these cells, with no positive expression in the nucleus. Conclusion: There were changes in the tissue morphology and structure of the cryptorchid testis, coupled with abnormally high expression of VEGF and P53 proteins in Erhualian pigs. We speculate that this may be an important limiting factor to fecundity during cryptorchidism. [ABSTRACT FROM AUTHOR]

Simple Summary: Deer farming for venison production has incorporated castration as a means to ease the management of male spikers (handling and injuries) and prevent breeding of slaughter animals. The present study assesses immunocastration as an animal-welfare-friendly alternative to physical castration methods, together with the use of supplementary amino acids to improve the nutrition of yearling fallow deer bucks. Indicators of testes development were assessed, as well as their correlations with secondary sexual traits and dimorphic body development (including antlers), which may be affected by castration and increased nutrition. Immunocastration disrupted testes development on a microscopic level (atrophy of seminiferous tubules, decreased sperm viability, and decreased percentages of normal sperm morphology), but testes size remains a questionable indicator of vaccination success on the macroscopic level. When the effect of nutrition is considered, amino acid supplementation shows potential for enhancing the development of yearling fallow deer bucks selected for breeding, as intact deer fed amino acids had the greatest extent of testes development and sperm viability. Thus, both aspects warrant further investigation as promising tools for application in deer farming. Forty-four fallow deer bucks (10 months old; 22.9 ± 2.4 kg) were utilized to investigate the effects of immunocastration and amino acid supplementation on testes development. Immunocastrated bucks were administered Improvac® at weeks 1, 8, and 20 of this study (control group: intact males). Starting at week 8, half of each sex received rumen-protected lysine and methionine (3:1) supplementation. At slaughter (week 37/39), body size, internal fat deposits, antler size parameters, testes weight, testes surface color, cauda epididymal sperm viability and morphology, and seminiferous tubule circumference and epithelium thickness were determined. Animals with larger body sizes, greater forequarter development, and antler growth also had greater testes development. Whilst the result of immunocastration on testes size is unexpected, testes tissue showed impaired development (atrophied seminiferous tubules), decreased sperm viability, and normal morphology. Testes tissue from immunocastrated deer was less red, possibly indicating reduced blood supply. Conversely, amino acid supplementation increased testes' redness and sperm viability, and intact males fed amino acids showed the greatest seminiferous tubule development. Thus, immunocastration may be a welfare-friendly alternative for venison production. Whilst the results support findings from the literature that testes size is not a reliable indicator of immunocastration success, this warrants further investigation in deer over different physiological development stages. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEA) is present worldwide as a serious contaminant of food and feed and causes male reproductive toxicity. The implication of paraptosis, which is a nonclassical paradigm of cell death, is unclear in ZEA-induced male reproductive disorders. In this study, the toxic effects of ZEA on the blood-testis barrier (BTB) and the related mechanisms of paraptosis were detected in goats. ZEA exposure, in vivo, caused a significant decrease in spermatozoon quality, the destruction of seminiferous tubules, and damage to the BTB integrity. Furthermore, ZEA exposure to Sertoli cells (SCs) in vitro showed similar dysfunction in structure and barrier function. Importantly, the formation of massive cytoplasmic vacuoles in ZEA-treated SCs corresponded to the highly swollen and dilative endoplasmic reticulum (ER), and paraptosis inhibition significantly alleviated ZEA-induced SC death and vacuolization, which indicated the important contribution of paraptosis in ZEA-induced BTB damage. Meanwhile, the expression of ER stress marker proteins was increased after ZEA treatment but decreased under the inhibition of paraptosis. The vacuole formation and SC death, induced by ZEA, were remarkably blocked by ER stress inhibition. In conclusion, these results facilitate the exploration of the mechanisms of the SC paraptosis involved in ZEA-induced BTB damage in goats. [ABSTRACT FROM AUTHOR]