Contractions transport exfoliated epithelial cells through the neonatal epididymis

Contractions of the adult epididymal duct are well known in the context of sperm transport. Some reports also describe contractions of the epididymal duct during development, but data about their character, regulation and function are sparse. In the foetal human epididymis we found luminal cells and could identify them as exfoliated epithelial cells originating from the epididymis and not from testis by using antibodies against neutral endopeptidase as an epithelial epididymal duct marker. Exfoliated cells were also found in the epididymal duct after birth. Time-lapse imaging revealed directional transport of luminal cells in the neonatal rat epididymis interrupted by pendular movement. Spontaneous contractions were discovered in the neonatal epididymis and an association between these contractions and the transport of the luminal cells could be observed. Both, transport and spontaneous contractions, were affected significantly by substances known to contract (noradrenaline) or relax (the phosphodiesterase 5 inhibitor sildenafil) smooth muscle cells. Immunohistochemistry showed staining for the proliferation marker proliferating-cell-nuclear-antigen (PCNA) in cells of the ductal lumen of the neonatal rat epididymis indicating the extrusion of cells also during proliferation. Our data showed spontaneous contractions of the immature epididymal duct associated with the transport of exfoliated luminal cells before the first occurrence of sperm cells. Results suggest an important role including both (i) a mechanical place holder function of exfoliated luminal cells (ii) together with a novel idea of organized waste disposal of these cells during development.

Morphophysiology of the Epididymis of the African Sideneck Turtle (Pelusios ‎castaneus): Histological, Microstereological and Ultrastructural Approach

This study was carried out to describe the morphophysiology of the epididymis of the adult African sideneck turtle using histological, microstereological and ultrastructural methods. The epididymal duct lies within a relatively thin sheath of connective tissue, and is lined by pseudostratified columnar epithelium. Unlike luminal diameter and stereocilial height, epithelial height, as well as the population of principal cells, decreased from the proximal to posterior segment. The clear cells of the turtle epididymis are limited to the posterior segment of the duct. Basal and apical cells as well as intra-epithelial lymphocytes are all distributed across the three segments of the epididymis while macrophage-like cells are absent throughout the length of the duct epithelium. The structure of the African sideneck turtle epididymis demonstrates, as in most mammals and few reptiles studied to date, obvious regional differentiation of the duct epithelium with evidences of secretory and endocytotic abilities as demonstrated by the contents of highly developed endoplasmic reticulum and secretory blebs in the principal and basal cells as well as clear cells, believed to be concerned with endocytosis. The outcome of the study is expected to be useful in the comparative structural and functional anatomy of turtle epididymis.

Immunohistochemical Studies of αSMA in the Epididymis of African Four-Toed Hedgehog (Atelerix albiventris)

The epididymis plays an important role in sperm maturation, storage, transport and in the secretion of enzymes and proteins into the tubular lumen. In this study, we examined the histology, microstereology and immunohistochemical localization of alpha smooth muscle (αSMA) in the three regions of the epididymis of the African four-toed hedgehog (Atelerix albiventris). Ten adult males were captured from the wild in Ibadan, Nigeria, between May and October, 2016. The animals were euthanized and the epididymis (caput, corpus and cauda regions) were retrieved and fixed in buffered neutral formalin ahead of the paraffin technique, following standard procedures. The duct of the epididymis was lined by pseudostratified columnar epithelium comprising basal, principal and apical cells as well as intraepithelial lymphocytes in proximity to basal cells. The principal cells, the major cells encountered within the epididymal epithelium of the animal, decreased in population from the caput to the cauda epididymidis while the apical cells were more abundant in the cauda epididymidis. Positive reactions to αSMA were observed in the peritubular muscular coat of the epididymal duct as well as blood vessels across the three regions of the epididymis with the caput and cauda epididymidis showing stronger positive reactions compared to the corpus epididymidis. This study demonstrated that the histology, microstereology as well as the cellular constituents of the epididymal duct of the Atelerix albiventris are similar to those of other mammals with a slight variation. It has also highlighted variation in the localization of αSMA across the regions of the epididymis of the animal.

Dexamethasone improves therapeutic outcomes in a preclinical bacterial epididymitis mouse model

STUDY QUESTION Can dexamethasone improve infertility-related cauda epididymidal tissue damage caused by bacterial epididymitis? SUMMARY ANSWER Dexamethasone in addition to anti-microbial treatment effectively reduces long-term deleterious epididymal tissue damage by dampening the host’s adaptive immune response. WHAT IS KNOWN ALREADY Despite effective anti-microbial treatment, ~40% of patients with epididymitis experience subsequent sub- or infertility. An epididymitis mouse model has shown that the host immune response is mainly responsible for the magnitude of epididymal tissue damage that is fundamentally causative of the subsequent fertility issues. STUDY DESIGN, SIZE, DURATION Bacterial epididymitis was induced in male mice by using uropathogenic Escherichia coli (UPEC). From Day 3 after infection onwards, mice were treated with daily doses of levofloxacin (20 mg/kg, total n = 12 mice), dexamethasone (0.5 mg/kg, total n = 9) or both in combination (total n = 11) for seven consecutive days. Control animals were left untreated, i.e. given no interventional treatment following UPEC infection (total n = 11). Half of the animals from each group were killed either at 10 or 31 days post-infection. PARTICIPANTS/MATERIALS, SETTING, METHODS A mouse model of induced bacterial epididymitis was applied to adult male C57BL/6J mice. At the respective endpoints (10 or 31 days post-infection), epididymides were collected. Effectiveness of antibiotic treatment was assessed by plating of epididymal homogenates onto lysogeny broth agar plates. Overall tissue morphology and the degree and nature of tissue damage were assessed histologically. Quantitative RT-PCR was used to assess local cytokine transcript levels. Blood was drawn and serum analysed for systemic IgG and IgM levels by ELISA. In addition, correlation analyses of clinical data and serum-analyses of IgG and IgM levels in patients with epididymitis were performed. MAIN RESULTS AND THE ROLE OF CHANCE The addition of dexamethasone to the standard anti-microbial treatment did not further worsen epididymal tissue integrity. In fact, an obviously dampened immune response and reduced tissue reaction/damage was observed at both 10 and 31 days post-infection following combined treatment. More specifically, epididymal duct continuity was preserved, enabling sperm transit. In contrast, in untreated or antibiotic-treated animals, damage of the epididymal duct and duct constrictions were observed, associated with a lack of cauda spermatozoa. In line with the bacteriostatic/bactericidal effect of levofloxacin (alone as well as in combination), local cytokine transcript levels were significantly and similarly reduced in animals treated with levofloxacin alone (P < 0.01) or in combination with dexamethasone (P < 0.05) compared to UPEC-infected untreated animals. Interestingly, the addition of dexamethasone to the anti-microbial treatment induced a unique dampening effect on adaptive immunity, since systemic IgG and IgM levels as well as the pan-T cell marker CD3 were reduced at both 10 and 31 days post-infection. LIMITATIONS, REASONS FOR CAUTION Breeding studies to address the fertility-protecting effect of the combined treatment were not possible in the experimental animals because the vas deferens was ligated (model specific). WIDER IMPLICATIONS OF THE FINDINGS Whereas innate immunity is necessary and involved in acute bacterial clearance, adaptive immunity seems to be responsible for long-term, subclinical immunological activities that may negatively affect the pathogenesis of bacterial epididymitis even after effective bacterial eradication. These effects can be reduced in mice by the additional treatment with dexamethasone. This immunological characteristic of bacterial epididymitis shows similarities to the Jarisch–Herxheimer reaction known from other types of bacterial infection. STUDY FUNDING/COMPETING INTEREST(S) The study was supported by grants from the Deutsche Forschungsgemeinschaft, Monash University and the Medical Faculty of Justus-Liebig University to the International Research Training Group on ‘Molecular pathogenesis of male reproductive disorders’ (GRK 1871). R.W., K.L.L. and M.P.H. were supported by grants from the National Health and Medical Research Council of Australia (ID1079646, ID1081987, ID1020269 and ID1063843) and by the Victorian Government’s Operational Infrastructure Support Program. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER No clinical trial involved.

58 Histomorphological Study of Epididymis, ductus deferens and phallus in Adult Male Guinea Fowl (Numida meleagris)

This study was conducted to investigate the anatomical, histological characteristic features of epididymis, ductus deferens and phallus of male guinea fowl (Numidia meleagris ). Fifteen healthy male guinea fowl aged between 18-24 months, weighing 1280 - 1405 gram were used in current study which brought from local market in Baghdad governorate. The epididymis of guinea fowl consisted from rete testis, efferent ductless, connecting duct and epididymal duct. The study revealed that the mean diameter of epididymal ducts was 285.17± 2.43µm lined with pseudostratified columnar epithelium. The epididymal duct continuous with ductus deferens which become wider caudally towards the cloaca, the length of right and left ductus deferens were 9.16±1.20 cm and 9.80±1.24cm respectively. The copulatory organ (phallus) of guinea fowl was not well developed and non-intermittent type consist from right and left lateral lymphoid bodies with median eminences, the transverse width 5.01±0.24mm, the cranio-caudal width 3.51±0.07mm and dorso- ventral width 4.32±0.25mm. The lining epithelium of coprodeum was pseudo stratified columnar epithelium. This study concluded that the epididymis of male guinea fowl located on dorsomedial surface of the testes and without definite parts as in domestic animals, the ductus deferens of guinea fowl was tortuous, the phallus of guinea fowl is non - intermittent type.

Contractility of the epididymal duct - function, regulation and potential drug effects

During their transit through the epididymis, spermatozoa mature and acquire motility and fertilizing capacity. The smooth muscle cells (SMCs) of the epididymal duct are thought to be responsible for the adequate transport of spermatozoa. Thus, precise regulation of SMC function also represents a prerequisite for sperm maturation thereby contributing to male fertility. In this review we would like to highlight various aspects of epididymal SMC function and discuss several angles with respect to regulation of contraction and relaxation. Different to the vas deferens, where disturbed SMC pathways resulting in male infertility could be defined, comparable information is missing in the epididymis. We therefore include some vas deferens data which could also be useful for a better understanding of epididymal SMC function. Furthermore, we would like to draw attention to drugs used in clinical practice and their potential (side) effects on contractions in the epididymis.