Epididymal Epithelium Research Articles

Isolation of CD63-positive epididymosomes from human semen and its application in improving sperm function.

Extracellular vesicles (EVs) are highly heterogeneous, and different EV subpopulations from various origins mediate different biological effects. The separation of different subpopulations of EVs from mixtures is critical but challenging. Epididymosomes are secreted by the epididymal epithelium and play a key role in sperm maturation and function. However, limited access to human epididymal tissue and epididymal fluid has hampered further study of epididymosomes and their potential clinical applications. Here, we established a novel strategy based on flow cytometry sorting to isolate human CD63-positive epididymosomes from ejaculate. We identified CD52, a membrane-located protein expressed exclusively in the epididymis, as the sorting marker for human epididymosomes. Then, CD63-positive epididymosomes were isolated from human semen using a flow cytometry sorting instrument and concentrated. Additionally, we observed that isolated CD63-positive epididymosomes improved sperm function more than other CD63-positive seminal EV subpopulations did, demonstrating the successful isolation of a subpopulation of epididymosomes from human semen and their potential application in the clinic.

Seasonal Variation in TLR4 Expression in The Testis and Epididymis of Anatolian Ground Squirrels (Spermophilus xanthoprymnus): Insights from Non-Breeding Period of Pre-Hibernation and Hibernation

This study investigates the modulation of Toll-like receptor 4 (TLR4) expression within the testis and epididymis of Anatolian ground squirrels (Spermophilus xanthoprymnus) during the non-breeding period of pre-hibernation and hibernation. Immunohistochemical investigation showed that TLR4 was not detected in germ cells, Leydig cells, or Sertoli cells in the testicular tissue during the pre-hibernation period. Nevertheless, there was a presence of TLR4 in the vessels walls and certain interstitial cells within the intertubular regions. Epithelial cells in the caput, corpus, and cauda regions of the epididymis showed no TLR4 expression. However, it was observed in the vessel walls, smooth muscle layers, and some interstitial cells. TLR4 expression was seen in spermatogonia and primary spermatocytes during hibernation, with strong labeling observed in the vessel walls of the intertubular area. In contrast, TLR4 was detected in the epididymal epithelium, as well as in the smooth muscle layers and vessel walls throughout all segments. A notable upregulation in the expression of TLR4 in the testis was identified through quantitative image analysis during hibernation as compared to pre-hibernation. During pre-hibernation, the cauda segment of the epididymis exhibited the highest expression of TLR4, whereas during hibernation, the corpus segment demonstrated the highest expression. These findings suggest a dynamic modulation of TLR4 in response to hibernation, highlighting its potential role in reproductive function and immune adaptation.

Advancement and Potential Applications of Epididymal Organoids.

The epididymis, a key reproductive organ, is crucial for sperm concentration, maturation, and storage. Despite a comprehensive understanding of many of its functions, several aspects of the complex processes within the epididymis remain obscure. Dysfunction in this organ is intricately connected to the formation of the microenvironment, disruptions in sperm maturation, and the progression of male infertility. Thus, elucidating the functional mechanisms of the epididymal epithelium is imperative. Given the variety of cell types present within the epididymal epithelium, utilizing a three-dimensional (3D) in vitro model provides a holistic and practical framework for exploring the multifaceted roles of the epididymis. Organoid cell culture, involving the co-cultivation of pluripotent or adult stem cells with growth factors on artificial matrix scaffolds, effectively recreates the in vivo cell growth microenvironment, thereby offering a promising avenue for studying the epididymis. The field of epididymal organoids is relatively new, with few studies focusing on their formation and even fewer detailing the generation of organoids that exhibit epididymis-specific structures and functions. Ongoing challenges in both clinical applications and mechanistic studies underscore the importance of this research. This review summarizes the established methodologies for inducing the in vitro cultivation of epididymal cells, outlines the various approaches for the development of epididymal organoids, and explores their potential applications in the field of male reproductive biology.

Inhibition of Prolactin Affects Epididymal Morphology by Decreasing the Secretion of Estradiol in Cashmere Bucks.

Yanshan Cashmere bucks are seasonal breeding animals and an important national genetic resource. This study aimed to investigate the involvement of prolactin (PRL) in the epididymal function of bucks. Twenty eleven-month-old Cashmere bucks were randomly divided into a control (CON) group and a bromocriptine (BCR, a prolactin inhibitor, 0.06 mg/kg body weight (BW)) treatment group. The experiment was conducted from September to October 2020 in Qinhuangdao City, China, and lasted for 30 days. Blood was collected on the last day before the BCR treatment (day 0) and on the 15th and 30th days after the BCR treatment (days 15 and 30). On the 30th day, all bucks were transported to the local slaughterhouse, where epididymal samples were collected immediately after slaughter. The left epididymis was preserved in 4% paraformaldehyde for histological observation, and the right epididymis was immediately preserved in liquid nitrogen for RNA sequencing (RNA-seq). The results show that the PRL inhibitor reduced the serum PRL and estradiol (E2) concentrations (p < 0.05) and tended to decrease luteinizing hormone (LH) concentrations (p = 0.052) by the 30th day, but no differences (p > 0.05) occurred by either day 0 or 15. There were no differences (p > 0.05) observed in the follicle-stimulating hormone (FSH), testosterone (T), and dihydrotestosterone (DHT) concentrations between the two groups. The PRL receptor (PRLR) protein was mainly located in the cytoplasm and intercellular substance of the epididymal epithelial cells. The PRL inhibitor decreased (p < 0.05) the expression of the PRLR protein in the epididymis. In the BCR group, the height of the epididymal epithelium in the caput and cauda increased, as did the diameter of the epididymal duct in the caput (p < 0.05). However, the diameter of the cauda epididymal duct decreased (p < 0.05). Thereafter, a total of 358 differentially expressed genes (DEGs) were identified in the epididymal tissues, among which 191 were upregulated and 167 were downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that ESR2, MAPK10, JUN, ACTL7A, and CALML4 were mainly enriched in the estrogen signaling pathway, steroid binding, calcium ion binding, the GnRH signaling pathway, the cAMP signaling pathway, and the chemical carcinogenesis-reactive oxygen species pathway, which are related to epididymal function. In conclusion, the inhibition of PRL may affect the structure of the epididymis by reducing the expression of the PRLR protein and the secretion of E2. ESR2, MAPK10, JUN, ACTL7A, and CALML4 could be the key genes of PRL in its regulation of epididymal reproductive function.

Exploring the Molecular Characteristics and Role of PDGFB in Testis and Epididymis Development of Tibetan Sheep.

Platelet-derived growth factor B (PDGFB), as an important cellular growth factor, is widely involved in the regulation of cellular events such as cell growth, proliferation, and differentiation. Although important, the expression characteristics and biological functions in the mammalian reproductive system remain poorly understood. In this study, the PDGFB gene of Tibetan sheep was cloned by RT-PCR, and its molecular characteristics were analyzed. Subsequently, the expression of the PDGFB gene in the testes and epididymides (caput, corpus, and cauda) of Tibetan sheep at different developmental stages (3 months, 1 year, and 3 years) was examined by qRT-PCR and immunofluorescence staining. A bioinformatic analysis of the cloned sequences revealed that the CDS region of the Tibetan sheep PDGFB gene is 726 bp in length and encodes 241 amino acids with high homology to other mammals, particularly goats and antelopes. With the increase in age, PDGFB expression showed an overall trend of first decreasing and then increasing in the testis and epididymis tissues of Tibetan sheep, and the PDGFB mRNA expression at 3 months of age was extremely significantly higher than that at 1 and 3 years of age (p < 0.05). The PDGFB protein is mainly distributed in testicular red blood cells and Leydig cells in Tibetan sheep at all stages of development, as well as red blood cells in the blood vessel, principal cells, and the pseudostratified columnar ciliated epithelial cells of each epididymal duct epithelium. In addition, PDGFB protein expression was also detected in the spermatocytes of the 3-month-old group, spermatids of the 1-year-old group, spermatozoa and interstitial cells of the 3-year-old group, and loose connective tissue in the epididymal duct space in each developmental period. The above results suggest that the PDGFB gene, as an evolutionarily conserved gene, may play multiple roles in the development and functional maintenance of testicular cells (such as red blood cells, Leydig cells, and germ cells) and epididymal cells (such as red blood cells, principal cells, and ciliated epithelial cells) during testicular and epididymal development, which lays a foundation for the further exploration of the mechanisms by which the PDGFB gene influences spermatogenesis in Tibetan sheep.

Cellular mechanism underlying leptin-induced anion secretion of rat epididymal epithelial cells.

A large number of studies have shown that leptin plays an important role in the regulation of fertility via the hypothalamus-pituitary-gonad axis. However, its peripheral function in epididymis was still elusive. The purpose of this study was to determine the pro-secretion effect of leptin on the rat epididymal epithelium. In the present study, real-time quantitative polymerase chain reaction, western blot, and immunohistochemical analysis were employed to detect the expression pattern of leptin receptors in rat epididymis. The pro-secretion effect of leptin on epididymal epithelial cells was measured by short-circuit current, and the prostaglandin E2 and cyclic adenosine monophosphate level was evaluated by enzyme-linked immunosorbent assay. We verified that the leptin receptor was located on the epididymal epithelium, with a relatively high expression level in corpus and cauda epididymis. Ussing chamber experiments showed that leptin stimulated a significant rise of the short-circuit current in rat epididymal epithelial cells, which could be abolished by the specific leptin receptor antagonist peptide Allo-aca, or by removing the ambient Cl- and HCO3 -. Furthermore, the leptin-stimulated short-circuit current response could be abrogated by blocking the apical cystic fibrosis transmembrane regulator or the basolateral Na+-K+-2Cl- cotransporter. Our pharmacological experiments manifested that interfering with the prostaglandin H synthase-2-prostaglandin E2-EP2/EP4-adenylate cyclase pathways could significantly blunt the cystic fibrosis transmembrane regulator-mediated anion secretion induced by leptin. The enzyme-linked immunosorbent assay demonstrated that leptin could induce a substantial increase in prostaglandin E2 release and cyclic adenosine monophosphate synthesis of primary cultured rat cauda epididymal epithelial cells. Our data also suggested that JAK2, ERK, and PI3K-dependent phosphorylation may be involved in the activation of prostaglandin H synthase-2 and the subsequent prostaglandin E2 production. The present study demonstrated the pro-secretion function of leptin in rat epididymal epithelium via the activation of cystic fibrosis transmembrane regulator and Na+-K+-2Cl- cotransporter, which was dependent on the paracrine/autocrine prostaglandin E2 stimulated EP2/EP4-adenylate cyclase pathways, and thus contributed to the formation of an appropriate microenvironment essential for sperm maturation.

Male genital system of Ameiva ameiva (Squamata: Teiidae).

Understanding squamate reproductive morphology is crucial for investigating ecological, behavioral, and evolutionary questions. Here, we describe the anatomy and histology of the male genital system of Ameiva ameiva from southeastern Brazil. Ten adult males were dissected to characterize genital macroscopy and collect fragments of the testes, gonadoducts, and kidneys for histological examination. We examined 10 transverse histological sections per individual and measured the epithelial height of the epididymis and ductus deferens. The male reproductive system consists of a pair of yellowish oval testes, the rete testis, ductuli efferentes, epididymis, ductus deferens, ampulla ductus deferentis, sexual segment of the kidney (SSK), cloaca, and hemipenis. The hemipenis is elongated, cylindrical, and unilobed, with a sulcate face and an asulcate face, which has continuous fringes throughout its length. Seminiferous tubules exhibited germ cells at various stages. The epididymis is wider and more coiled than the ductus deferens. The rete testis has a simple squamous epithelium with long stereocilia, while the narrower ductuli efferentes are lined by a simple ciliated cuboidal epithelium. The epididymal epithelium is pseudostratified columnar, with basal and ciliated principal cells, whereas the ductus deferens epithelium is pseudostratified to simple cuboidal. The epididymal epithelium is 1.5 times taller than the ductus deferens epithelium. Here, we observed the SSK present in the cortex of the ventral region of the kidneys due to the hypertrophy of the distal convoluted tubules, as well as its secretory activity. Our findings will contribute to future research into the evolution of squamate reproductive morphology.

RUNX transcription factors are essential in maintaining epididymal epithelial differentiation

Apart from the androgen receptor, transcription factors (TFs) that are required for the development and formation of the different segments of the epididymis have remained unknown. We identified TF families expressed in the developing epididymides, of which many showed segment specificity. From these TFs, down-regulation of runt related transcription factors (RUNXs) 1 and 2 expression coincides with epithelial regression in Dicer1 cKO mice. Concomitant deletion of both Runx1 and Runx2 in a mouse epididymal epithelial cell line affected cell morphology, adhesion and mobility in vitro. Furthermore, lack of functional RUNXs severely disturbed the formation of 3D epididymal organoid-like structures. Transcriptomic analysis of the epididymal cell organoid-like structures indicated that RUNX1 and RUNX2 are involved in the regulation of MAPK signaling, NOTCH pathway activity, and EMT-related gene expression. This suggests that RUNXs are master regulators of several essential signaling pathways, and necessary for the maintenance of proper differentiation of the epididymal epithelium.

Maternal protein restriction affects the differentiation of cells in the epididymal epithelium lining of 44-day-old rats.

Maternal protein restriction delays the differentiation of epididymal mesenchymal cells in newborn rats. However, it's unclear if this delay persists until the full differentiation of the epididymal epithelium at 44 days postnatal. Thus, this study aimed to assess the impact of maternal protein reduction on 44-day-old rats' epididymal epithelium differentiation, following up on the observed delay in newborn animals. Pregnant rats were randomly divided into groups receiving normal-protein (NP - 17% protein) or low-protein (LP - 6% protein) diets during gestation and lactation. On postnatal day (PDN) 44, male offspring were euthanized, and the epididymis (NP n=10, LP n=10) was processed according to immunohistochemical techniques for the detection of aquaporin 9 (AQP9), KI-67, TP63, and ATPase. LP rats showed: a decrease in the intensity of the AQP9 reaction, an increase in cellular proliferation in the initial segment and corpus of the epididymis, an increase in basal cells in the caput and corpus epididymis, and an increase in ATPase-positive clear cells in the cauda region. These findings demonstrate that maternal protein restriction impacts cell differentiation in the epididymal epithelium of 44-day-old rats, persisting even with a normal-protein diet after weaning.

Three-Dimensional Cell Culture of Epididymal Basal Cells and Organoids: A Novel Tool for Toxicology.

Spermatozoa are formed in the testis but must transit through the epididymis to acquire motility and the ability to fertilize. The epididymis is a single convoluted tubule comprising several anatomically and physiologically distinct regions. The pseudostratified epithelium consists of multiple cell types, including principal cells, clear cells, narrow cells, and apical cells, that line the lumen of the epididymis. Basal cells are present at the base of the epithelium, and halo cells, which includes macrophages/monocytes, mononuclear phagocytes, and T lymphocytes, are also present in the epithelium. Several aspects of this complex spermatozoan maturation process are well established, but a great deal remains poorly understood. Given that dysfunction of the epididymis has been associated with male infertility, in vitro tools to study epididymal function and epididymal sperm maturation are required. Our lab and others have previously developed human, rat, and mouse epithelial principal cell lines, which have been used to address certain questions, such as about the regulation of junctional proteins in the epididymis, as well as the toxicity of nonylphenols. Given that the epididymal epithelium comprises multiple cell types, however, a 3D in vitro model provides a more comprehensive and realistic tool that can be used to study and elucidate the multiple aspects of epididymal function. The purpose of this article is to provide detailed information regarding the preparation, maintenance, passaging, and immunofluorescent staining of rat epididymal organoids derived from adult basal cells, which we have demonstrated to be a type of adult stem cell in the rat epididymis. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolation of epididymal cells Basic Protocol 2: Magnetic activated cell sorting and isolation of basal cells Basic Protocol 3: Preparation and culture of epididymal basal cell organoids Basic Protocol 4: Passage of epididymal basal cell organoids Basic Protocol 5: Freezing and thawing of epididymal basal cell organoids Basic Protocol 6: Immunofluorescent staining of epididymal basal cell organoids.

Morphometric changes in the gonads of the West African dwarf bucks experimentally infected with Trypanosoma vivax and Trypanosoma brucei and response due to diminazine aceturate treatment

Trypanosomosis, one of the major diseases affecting livestock production in Africa has been associated with reproductive disorders. The disease has not been extensively studied in goats especially the West African Dwarf (WAD) goat. The morphology of the gonads of WAD bucks experimentally infected with Trypanosoma (T.) vivax and Trypanosoma brucei and treatment with Diminazine aceturate were investigated in this study. The study lasted a period of 20 weeks, the first 3 weeks were used to acclimatize the animal to their new environment. The remaining 16 weeks were used to carry out the experiment. Two species of trypanosome were used for the experiment; T. brucei (NITR CT/28- Federe strain) and T. vivax (A field strain obtained from an abattoir). The experiment was divided into three stages tagged Pre-infection (A), infection (B) and treatment (C) while the animals were also divided into control group and infected groups- Trypanosoma brucei and Trypanosoma vivax groups. Biometric parameters that include testicular weight, height and others testicular and epididymal indices were measured according to standard techniques. Data were summarized with mean± SEM and analysed using ANOVA. There were no significant changes in body weight between the control group and treated groups with either T. brucei or T. vivax. The little reduction in scrotal circumference during infection and subsequent increase during post treatment were not significant (P &gt; 0.05) compared to the control. The Testicular weight, testicular length and epididymal weight were not significantly affected during infection with both T. brucei and T. vivax. Tubular diameter and epithelia height of the seminiferous tubules were significantly (P≤0.05) lower in the T. brucei and T. vivax infected groups compared to the control. On the other hand, the epididymal tubular diameter and epididymal epithelia height were not significantly (P&gt;00.05) affected by the treatment. The study showed that the fertility impairment sequel to trypanosomosis is possibly due to alteration in the biometric parameter of the testis.

Histological study of Bactrian camel cryptorchidism and expression of immunoglobulin λ light chain in the testicular and epididymis of cryptorchid Bactrian camel.

Lambda light chains (λ-LCs) are frequently responsible for triggering the activation of inflammatory factors in autoimmune disorders, and an increase in their levels will cause various pathological changes in serum. The aim of this study was to determine the histological differences between the epididymis and testis of normal and cryptorchid Bactrian camels and the differences in λ-LC expression in the epididymis and testis of normal and cryptorchid Bactrian camels. Haematoxylin and eosin (H&E) staining was used to examine the pathological changes in cryptorchidism. The gene and protein levels of λ-LC were determined using RT-qPCR and western blot. The distribution of λ-LCs was assessed by immunohistochemistry and immunofluorescence. Compared with that in normal Bactrian camels, the diameter of the epididymal lumen and the thickness of the epithelium were decreased in the epididymis of cryptorchidic animals. Additionally, no sperm was detected in the cavity of the cryptorchidic epididymis. Meanwhile, the expression of λ-LC was significantly increased in the cryptorchidic epididymis at both the mRNA and protein levels (p < .05). The highest protein expression of λ-LC was found in epididymal epithelial halo cells and testicular Sertoli cells. These findings suggested that the structural changes observed in the epididymal epithelium of cryptorchidic camels affect its secretory and absorptive functions. Additionally, the high level of λ-LC expression recorded in halo cells suggested that these cells play an important role in epithelial immunity in cryptorchidic Bactrian camels. Furthermore, the high λ-LC expression levels detected in normal testicular Sertoli cells indicated that λ-LCs may be involved in spermatogenesis. The results of this study provide clues for an in-depth study of immunological sterility in cryptorchidic Bactrian camels.

Rosa26 Locus Is Inactive in Spermatogenesis of C57BL/6 Mice

ABSTRACT Background: The mouse Rosa26 locus demonstrates ubiquitous transcriptional activity, encoding long noncoding RNAs (LncRNAs) with no functional significance in various types of tissues and cells. It is widely employed in targeting transgene knock-in studies. However, the expression pattern of Rosa26 in the testes remains elusive. In this study, we conducted a detailed transcriptional analysis of the Rosa26 locus during mouse spermatogenesis. Methods: The transcriptional activity of the Rosa26 locus in the testes was assessed through immunohistochemistry or direct fluorescence observation in two Rosa26 locus-targeting knock-in mouse lines, Rosa26-Flag-Cas9 and Rosa26-mT/mG, both with a C57BL/6 background. The two mouse lines feature constitutively expressed Flag-tagged Cas9 and red fluorescent membrane-tethered tdTomato (mTomato) proteins. Results: In Rosa26-Flag-Cas9 mice, immunostaining of testis sections revealed robust expression of the Flag-Cas9 transgene in spermatogonia adjacent to the basement membrane of seminiferous tubules. However, its expression was absent in germ cells undergoing spermatogenesis in seminiferous tubules and in maturing spermatids in the epididymis. The transgene was also observed in intertubular Leydig cells and epididymal epithelia but was absent in Sertoli cells. Similarly, in Rosa26-mT/mG mice, fluorescence microscopy showed a complete absence of mTomato expression in the spermatogonial lineage and Sertoli cells. However, robust mTomato expression was observed in intertubular Leydig cells out of the seminiferous tubules and epididymal epithelia. Conclusion: Our findings suggest that the Rosa26 locus is inactive during spermatogenesis, indicating that functional gene studies by targeting knock-in transgenes at the Rosa26 locus in the spermatogonial lineage may not be applicable in C57BL/6 mice.

Comparative Effects of Vitamin A and E Dietary Supplements on Semen Traits, Reproductive System Morphology and Vitamin Storage in Rooster

The purpose of this study was to determine the effect of long-term vitamin A and E dietary supplements on the semen traits and morphology of the reproductive system in older roosters to prolong their reproductive contributions to the flock. The research revealed that long-term dietary supplementation of both vitamins A and E maintained the traits of the semen at levels significantly higher than in the control roosters. Vitamin A especially maintained ejaculate volume and semen density, while vitamin E protected semen motility and viability. Vitamin A treatment maintained the uniformity of the epididymal epithelium. No eroded areas were identified in this epithelium. Most epithelial cells were principal cells with uniform stereocilia. The connective tissue which lines the epididymal tubule of the vitamin A treated rooster was thinner than those of the roosters from the control. Vitamin E predominantly protected the structure of the spermatogenesis line. Liver tissue concentrations of vitamins A and E were two times and four times higher respectively than the control. The two vitamins were eliminated in increased quantities through the seminal plasma in supplemented roosters. No mutual inhibition or potentiating effects were found between the two vitamins, suggesting that their combined administration should be further analyzed.

Seasonal variations of the epididymis in donkeys (Equus asinus) with special reference to blood epididymal barrier.

The epididymis, a key component of the male reproductive system, controls spermatozoa's maturation, fertility, and storage. The objective of this study is to evaluate the histological, ultrastructural, and immunohistochemical variations in the epididymis of donkeys that occur throughout the year. During the breeding season (spring) and nonbreeding seasons (summer, autumn, and winter), 20 epididymis were collected from adult, clinically healthy donkeys. Compared to non-breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season. The epithelial height is the lowest and the lumen is the widest during the breeding season. Furthermore, the epididymal epithelium in the tail region exhibits undulations with polyps-like projections. The epididymal epithelium is composed mainly of the principal, basal, and dark cells. Tight junction between adjacent principal cells is more obvious in the breeding season as compared to the non-breeding seasons. However, intraepithelial lymphocytes, phagocytic, and other immune cells are more frequent in non-breeding seasons. β-catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer. Additionally, during non-breeding seasons, autophagy was detected within the epididymal epithelium which may be linked to stress adaptation. In conclusion, our findings suggest that the histological and ultrastructural characteristics of the epididymal epithelium are more active during spring compared to other seasons of the year. RESEARCH HIGHLIGHTS: The study aimed to evaluate the histological, ultrastructural, and immunohistochemical variations in the blood epididymal barrier (BEB) and epididymal epithelium of donkeys that occur throughout the year. In comparison to non-breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season. The epithelial height is the lowest and the lumen is the widest during the breeding season. The epididymal epithelium in the tail region exhibits undulations with polyps-like projections that increase the surface area. β-catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer.

Histomorphometric study of seminiferous tubules and epididymis in adult ram

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

Immortalized mouse caput epididymal epithelial (mECap18) cell line recapitulates the in-vivo environment.

Residing between the testes and the vas deferens, the epididymis is a highly convoluted tubule whose unique luminal microenvironment is crucial for the functional maturation of spermatozoa. This microenvironment is created by the combined secretory and resorptive activity of the lining epididymal epithelium, including the release of extracellular vesicles (epididymosomes), which encapsulate fertility modulating proteins and a myriad of small non-coding RNAs (sncRNAs) that are destined for delivery to recipient sperm cells. To enable investigation of this intercellular communication nexus, we have previously developed an immortalized mouse caput epididymal epithelial cell line (mECap18). Here, we describe the application of label-free mass spectrometry to characterize the mECap18 cell proteome and compare this to the proteome of native mouse caput epididymal epithelial cells. We report the identification of 5,313 mECap18 proteins, as many as 75.8% of which were also identified in caput epithelial cells wherein they mapped to broadly similar protein classification groupings. Furthermore, key pathways associated with protein synthesis (e.g., EIF2 signaling) and cellular protection in the male reproductive tract (e.g., sirtuin signaling) were enriched in both proteomes. This comparison supports the utility of the mECap18 cell line as a tractable in-vitro model for studying caput epididymal epithelial cell function.

A mixture of glyphosate and 2,4-D herbicides enhances the deleterious reproductive outcomes induced by Western diet in obese male mice.

The consumption of Western diet (WD) - enriched in fats and sugars - is associated with overweight, obesity and male reproductive disorders. In addition to WD intake, crops and dairy products display residues of herbicides, including glyphosate and 2,4-D that are widely applied worldwide. The concomitant exposure to WD and herbicides - mimicking contemporary scenarios - is not fully investigated. Thus, we evaluated the effects of glyphosate and 2,4-D, alone or in mixture, on WD-induced alterations in the male genital system. Male C57BL6J mice were submitted to WD (chow containing 20% lard, 0.2% cholesterol, 20% sucrose, and high sugar solution with 23.1 and 18.9 g/L of D-fructose and D-glucose) for 6 months. Concomitantly to WD, the animals received glyphosate (0.05, 5, or 50 mg/kg/day), 2,4-D (0.02, 2 or 20 mg/kg/day) or their mixture (0, 05 + 0.02, 5 + 2, or 50 + 20 mg/kg/day) by intragastrical administration (5×/week). Doses were based on Acceptable Daily Intake (ADIs) or No Observed Adverse Effect Level (NOAEL) values. Herbicide exposure did not alter the WD-induced obesity, hypercholesterolemia and hyperglycemia. WD induced sperm cell abnormalities, reduced the number, volume and area of Leydig cells, enhanced the frequency of epididymal abnormalities, decreased the proliferation in both germinal and epididymal epithelia, and reduced the number of androgen receptor (AR) positive epididymal cells. Remarkably, the herbicide mixtures promoted such WD-induced effects: increased the frequency of sperm cell and epididymal abnormalities (absence of sperm, cytoplasmic vacuoles, and clear cell hypertrophy) (5 + 2 and 50 + 20 doses); decreased Leydig cell nuclei volume and area (5 + 2 and 50 + 20 doses), reduced epididymal cell proliferation (all mixtures), and AR expression (50 + 20 dose). In addition, herbicide mixtures reduced serum testosterone levels (5 + 2 and 50 + 20 doses). Our findings indicate that the mixture of glyphosate and 2,4-D herbicides, mimicking environmentally relevant scenarios, promotes WD-induced changes in the male genital system.

Evaluation of the probable alleviating effect of luteolin on acrylamide mediated toxicity of the caput epididymal lining epithelium in adult rats: Histological and biochemical study

Evaluation of the probable alleviating effect of luteolin on acrylamide mediated toxicity of the caput epididymal lining epithelium in adult rats: Histological and biochemical study

Protective roles of vitamin C and 5-aminosalicylic acid on reproduction in acrylamide intoxicated male mice

ContextSerious health risks have been connected to ongoing, escalating exposure to environmental toxins and one of them is acrylamide (ACR), an organic compound. Although there are many published reports on ACR toxicity, limited information is available regarding the use of two potential antioxidants against ACR-instigated reproductive toxicity. AimsThe study focused on investigating the protective effects of vitamin C and 5-ASA against ACR-incited reproductive toxicity. MethodsA total of 50 male mice aged 4 weeks old were treated for 90 days with different concentrations either of ACR or ACR and vitamin C or ACR and 5- ASA or ACR, vitamin C, and 5- ASA. Key resultsACR significantly reduced serum testosterone level (p = 0.0037), sperm concentration (p = 0.0004), and percentage of sperm motility (p = 0.003), as well as increased sperm abnormality; head (p = 0.0058), tail (p = 0.001), and midpiece (p = 0.0339). Besides, the weight (p = 0.0006) and length (p = 0.0105) of testes, as well as weight (p = 0.0001) and length (p = 0.0021) of epididymis were decreased along with atrophy of seminiferous tubules of the testis, and disintegration of the tubular epithelium of epididymis on ACR exposed mice which were improved by vitamin C and 5-ASA administration. ConclusionsVitamin C and 5-ASA can potentially mitigate the negative effects of ACR on male reproduction; however, combined application is recommended for better performance. ImplicationsIn Bangladesh, this work is anticipated to address the health benefits of vitamin C and 5-ASA, particularly in improving the reproductive health of males against ACR toxicity.