The surface features of the epithelial lining of the ducts of the epididymis of the ostrich (Struthio camelus)

The luminal appearance of the various ducts of the epididymis of the ostrich was studied by scanning electron microscopy in tissues fixed by immersion in glutaraldehyde. The ductal types were similar to those previously described for some other species of birds. Numerous short microvilli, as well as a single cilium, projected from the apical surface of the rete testis cell. The ciliated cells of the efferent ductules projected tufts of cilia into the ductal lumen, while the non-ciliated cells bore short microvilli. The connecting and epididymal ducts were lined by a columnar cell type whose apical surface bore uniformly distributed microvilli and a single, centrally situated cilium. The spermatozoa found in all ducts of the epididymis bore a distal cytoplasmic droplet. This observation has implications for the maturational process in the ostrich spermatozoon in the epididymis. The surface features of the ducts, except for a few noteworthy differences, were generally similar to those previously described for the male domestic fowl, turkey and duck.     

The Excurrent Ducts of the Testis of the Emu (Dromaius novaehollandiae) and Ostrich (Struthio camelus): Microstereology of the Epididymis and Immunohistochemistry of its Cytoskeletal Systems

The volumetric proportion of the various ducts of the epididymis of the emu and ostrich and the immunohistochemistry of actin microfilaments, as well as cytokeratin, desmin and vimentin intermediate filaments, were studied in the various ducts of the epididymis of the emu and ostrich. The volumetric proportions of various ducts, which are remarkably different from those of members of the Galloanserae monophyly, are as follows: the rete testis, 5.2 ± 1.4% for the emu and 2.4 ± 1.8% for the ostrich; efferent ducts, 14.2 ± 2.3% (emu) and 11.8 ± 1.8% (ostrich); epididymal duct unit, 25.8 ± 5.8% (emu) and 26.1 ± 4.1% (ostrich) and connective tissue and its content, 54.7 ± 5.8% (emu) and 60.0 ± 4.9% (ostrich). Unlike in mammals and members of the Galloanserae monophyly, only vimentin was immunohistochemically demonstrated in the rete testis epithelium of the emu, and none of the cytoskeletal protein elements in the ostrich rete testis. The epithelium of the efferent ducts of the emu co-expressed actin, cytokeratin and desmin in the non-ciliated type I cells, and vimentin in the ciliated cell component. The ostrich demonstrated only cytokeratin in this epithelium. The ratite epididymal duct unit is different from that of mammals in lacking actin (only weaky expression in the ostrich), desmin and cytokeratin, and a moderate/strong immunoexpression of vimentin in the basal cells and basal parts of the NC type III cell in the epididymal duct unit. Immunoexpression of the microfilaments and intermediate filaments varied between the two ratite birds, as has been demonstrated previously in birds of the Galloanserae monophyly, and in mammals.     

Epithelial response to experimentally introduced intraluminal bacteria in the avian epididymal ducts

The response of the epithelial cells of the various ducts of the avian epididymis, whose function is poorly understood, to intraluminal bacteria was evaluated by the injection of an avirulent strain of Salmonella gallinarium into the RT for 24 h. Ultrastructurally, bacteria and invading mononuclear cells were present in the lumina of the RT, proximal efferent ducts (PED) and distal efferent ducts. However, only the non-ciliated (Type I) cells of the PED epithelium ingested bacteria from the lumen. Fragments of bacteria also occurred in several intercellular spaces in the epithelium of the PED. Some mononuclear cells also contained fragments of bacteria. Neither cell death in the various epithelia nor mononuclear infiltration of the periductal tissue occurred. Therefore, in addition to the established function of absorbing most of the testicular fluid entering the epididymis, the Type I cells also appear capable of recognising and removing foreign particulate matter from the epididymal through-flow in the proximal part of the epididymis.     

An unusual mechanism of disposal of superfluous spermatozoa in vasectomised quails

As part of a series of studies designed to evaluate usual and potential functions of epithelial cells in the avian epididymis under varying circumstances, adult, sexually active male Japanese quails were vasectomised for 3, 6 and 8 weeks. The testes, epididymides and deferent ducts were studied histologically and ultrastructurally. Testicular lesions were minimal, but epididymal response was profound. Non-ciliated (type I) cells of the proximal efferent duct (PED) actively phagocytized spermatozoa and their fragments. An especially noteworthy and apparently novel observation was the proliferation of the lining epithelium in parts of the PED to form additional adluminal sheets of spermiophagic epithelium which did not lie directly on basal laminae. These sheets were composed exclusively of non-ciliated cells. Spermatozoal dissolution and macrophage activity were also important mechanisms for the disposal of superfluous and incarcerated spermatozoa in the epididymis of this bird.     

Active spermiophagy in the initial part of the proximal efferent duct of the epididymis of normal domestic fowl (Gallus domesticus)

In a study of the absorptive activities of the excurrent ducts of the testis of birds, six adult cocks of the Ovambo breed of domestic fowl were deeply anaesthetised and intravascularly perfused with buffered 3 per cent glutaraldehyde. Epididymal tissue was prepared conventionally for electron microscopy. In favourable sections in three of these birds avid ingestion of spermatozoa was revealed in the non-ciliated (Type I) cells of the epithelial lining of the initial part of the proximal efferent duct, at and a little beyond its junction with the rete testis. No obvious defects were detected in luminal spermatozoa. The testicular excurrent ducts were neither distended nor obstructed, and mononuclear cell mobilisation was absent in both the ductal and periductal tissue. The significance of this observation with regard to the specific segment of the efferent duct system involved and how certain spermatozoa are identified for elimination from the duct lumen, must await precise studies.     

Morphological and glycohistochemical studies on the epididymal region of the Sudani duck (Cairina moschata)

In this study, the epididymal region of the Sudani duck was investigated using histological and lectin histochemical methods. Morphologically, the epididymal region of the Sudani duck is composed of extratesticular rete testis, proximal and distal efferent ductules, a short connecting duct, and epididymal ducts. Morphometric analysis of the epididymal region of Sudani duck revealed that the efferent ductules predominate in relation to the epididymal ducts. The distribution of sugar moieties within the epididymal region of the Sudani duck was investigated using ten different fluorescein isothiocyanate (FITC) conjugated lectins. In the rete testis epithelium, only PHA-L showed a positive reaction. Efferent ductules in contrary exhibited a wide range of lectin affinity whereas six positive lectins (Con A, LCA, PNA, WGA, PHA-L, PHA-E) were observed. In the connecting and epididymal ducts, four lectins (Con A, WGA, PHA-L, PHA-E) were also detected. GSA-I, UEA-I, and LTA were at all not evident in the epididymal region of the Sudani duck. In conclusion, the correlation between the large areas of the epididymal region occupied by the efferent ductules and the wide range of sugar affinity of this portion may confirm the speculation that efferent ductules might be the primary site of fluid reabsorption in the epididymal region of Sudani duck.     

Cyclical reproductive changes in the non-ciliated epithelia of the epididymis of birds

The epididymis of two species of domestic birds, the domestic fowl (Gallus domesticus), duck (Anas platyrhynchos), and of domestic and feral guinea-fowl (Numida meleagris) was studied during the three main phases of the reproductive cycle (prepuberal, sexually mature and active, and sexually mature but inactive or resting) with a view to identifying major histological and ultrastructural changes associated with and distinctive for each phase. Rete testis cells accumulated numerous variably sized lipid droplets in all birds, as well as large heterogeneous and lipofuscin-containing dense bodies in the guinea-fowl, during the resting but not in the other phases. The principal or Type III cells of the connecting and epididymal ducts exhibited profound structural changes, including, but not limited to, rarefied cytoplasm, inconspicuous and general loss of sparsely granular endoplasmic reticulum, loss of secretory vesicles in the drake, and an enhanced and conspicuous presence of lipid droplets in the guinea-fowl. The rete cells appeared to be less sensitive than the Type III cells to a reduced level or absence of lumenal androgens. These phase-dependent changes may help to prevent or minimize discrepancies in the interpretation of the normal structure of the epididymis in birds during the sexually active phase, as distinct from the other two phases and their intermediate phases.     

Ultrastructural study on junctional complexes of the excurrent duct epithelia in the epididymal region in the fowl.

Junctional complexes of the epithelia lining the rete testis, efferent ductules, connecting ductules and epididymal duct in the fowl were examined by transmission electron microscopy and by a tracer method using lanthanum nitrate. The junctional complexes were composed of tight junctions, adhering junctions and desmosomes. In the rete testis, one or two points of membrane fusion were observed at the tight junctions. In the efferent and connecting ductules and epididymal duct, the tight junctions consisted of a series of punctate membrane fusions. The adhering junctions and desmosomes showed no remarkable structural differences among these excurrent ducts. Vascularly infused lanthanum nitrate penetrated into the tight junctions of individual epithelia for variable distances, but was prevented from entering the lumen at the site of membrane fusion. These results suggest that the tight junctions can restrict the diffusion of materials via the paracellular route, and that they play an important role in maintaining a suitable fluid environment within the excurrent ducts.     

Localization of Oestrogen Receptors in the Epididymis During Sexual Maturation of the Domestic Cat

Oestrogens are involved in regulation of spermatogenesis and sperm maturation and are essential for male fertility. To study the role of oestrogens on epididymal function in the domestic cat, we analyzed the localization patterns of oestrogen receptors (ERs) within the epididymis of juvenile, pubertal and adults using immunohistochemistry. Cat epididymal tissues obtained during routine castrations were fixed in chilled Bouin's solution and processed for immunohistochemistry with ER-specific antibodies. For a certain receptor type, ER localization was influenced by donor age. In the juvenile epididymis, ERα was localized in the nuclei of epithelial cells of efferent ducts and undifferentiated epithelium of the ductus epididymis. During puberty, ERα localization in the undifferentiated epithelium of the epididymis shifted from the nuclei to the cytoplasm and plasma membrane. Oestrogen receptor-α level was highest in the pubertal and adult epididymis, especially within the cytoplasm and in plasma membranes of caput epithelial cells. This finding was suggestive of a role in fluid reabsorption within the efferent ducts and the epididymis. In corpus and cauda regions, ERα was less abundant, suggesting a minor role for oestrogens in sperm storage areas. Interestingly, localization of ERβ was neither influenced by age nor location within the epididymis and was ubiquitous throughout. Results demonstrate that oestrogen actions within the epididymis may be predominantly mediated through ERα during sexual maturation in the domestic cat.     

Immunolocalization of Aquaporin Water Channels in the Domestic Cat Male Genital Tract

Four different aquaporins (AQP1, 2, 5 and 9), integral membrane water channels that facilitate rapid passive movement of water, were immuno‐localized in the excurrent ducts collected from sexually mature cats during orchiectomy. Aquaporins 1, 2 and 9, were immuno‐localized at distinct levels, whereas AQP5 was undetectable all along the feline genital tract. No immunoreactivity was present at the level of the rete testis with any of the antibodies tested. In the efferent ducts, AQP1‐immunoreactivity was strongly evidenced at the apical surface of the non‐ciliated cells, and AQP9‐immunoreactivity was shown at the periphery of both ciliated and non‐ciliated cells. Aquaporins 2 was absent in the caput epididymidis, either in the efferent ducts or in the epididymal duct. Otherwise, AQP2 was increasingly localized at the adluminal surface of principal cells from the corpus to the cauda epididymidis and more weakly in the vas deferens epithelium. The supranuclear zone of the epididymal principal cells was AQP9‐immunoreactive throughout the duct, with the exclusion of the vacuolated sub‐region of the caput and with higher reaction intensity in the cauda region. AQP1 was present in the blood vessels all along the genital tract. AQP1 was expressed also in the smooth muscle layer of the vas deferens. The tested AQP molecules showed a different expression pattern in comparison with laboratory mammals, primates and the dog, unique other carnivore species studied to date. The present information is possibly useful in regard to the regional morphology of the feline epididymis and correlated functions, which are still a matter of debate.     

Immunolocalization of sex steroid receptors in the epididymis and ductus deferens of immature and mature Japanese Quail, Coturnix Japonica

The goal of this study was to determine whether in the Japanese quail the male genital tract contains receptors for progesterone, androgen and estrogen (PR, AR and ER, respectively), which have significant roles in reproductive functions, and whether their localization changes during sexual maturation. The epididymis and ductus deferens (middle and ampulla regions) of immature (approximately 30-day-old) and mature male Japanese quail were collected and frozen sections of them were immunostained for PR, AR and ER. The immunoreaction products for AR and PR were found in the nuclei of epithelial cells in the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens of mature and immature birds. In the mature birds, the epithelial cells of the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens were positive for ER, although some of the cells in the ductus deferens were negative. The epithelial cells of the ductules in the epididymis stained positive for ER, but the immunoreactions were negligible in the ductus deferens of immature birds. These results suggest that the epididymis and ductus deferens in quail possesses PR, AR and ER receptors. Each receptor is expressed before sexual maturation, although enhancement of ER expression may occur during maturation.     

Morphological features of the luminal surface of the magnum in the sexually immature ostrich (Struthio camelus)

Summary Observations were made, using scanning electron microscopy, of the surface features of the magnum in the immature ostrich during periods of ovarian inactivity, activity and regression. In birds with inactive ovaries the luminal surface of the magnum was lined with non-ciliated cells, which were densely covered by microvilli. In contrast, the magnum in birds with active ovaries was composed of ciliated and non-ciliated cells. The distribution of ciliated cells was not uniform, with clumps of cilia occurring next to non-ciliated areas. Samples collected from birds with regressing ovaries, during periods of decreasing daylength, revealed that the magnum was undergoing involution. The deciliation of ciliated cells and the presence of short microvilli on non-ciliated cells characterized magnal regression. These results suggest that ovarian activity and changes in daylength have a profound effect on the surface features of the magnum in the immature ostrich.     

The vascularization of the epididymis of the boar with special reference to perfusion fixation

Vasculature of the epididymis was investigated by means of corrosion casts. In the boar, epididymal arteries form a complex network around their stem vessel, the testicular artery. Proper perfusion fixation or complete casting therefore require direct injection into one of these branches. To reach the distal cauda, cannulation of the deferential artery is further needed. Connections between all of these feeding vessels occur at the level of the vascular cone. A prominent anastomosis between an epididymal branch and the testicular artery is regularly observed under the caput epididymis. Epididymal veins drain to a large extent into the pampiniform plexus. Unlike the situation in other species, vascularization of testis and epididymis are closely associated in the boar.     

Scanning electron and light microscopic studies of the surface epithelium of the rete testis and epididymis of the boar. I. Rete testis and efferent ducts

The use of the scanning electron microscope gave a three dimensional representation of the epithelial surface. Additionally, light microscopy revealed the representative structure of the epithelium. The rete testis showed a single layer of cubic epithelial cells. Short and dense microvilli were found on the surface. Sporadically a single, cilia-like structure was recognized. An extratesticular rete testis was identified. The flowing transition of the epithelium between the rete testis and the efferent ductuli occurred at different levels, so that both kinds of epithelial structures were recognized in the same area. The efferent ductuli were composed of a single columnar epithelium consisting of two cell types, principal cells and ciliated cells. The ciliated cells were recognized by their cilia protruding into the lumen. The principal cells showed microvilli on their surface and bleblike apical protrusions which erupt into the lumen.     

Morphology and innervation pattern of the feline urogenital junction

The feline urogenital junction is situated between the extratesticular rete and the spacious initial segments of the efferent ductules. The rete epithelium is cuboidal to low columnar. The rete cells forming the junction rest on a wavy basal lamina, display deep mutual invaginations, possess central nuclei with several infoldings and form a distinct border with the columnar epithelial cells of the initial segments of the ductuli efferentes. The epithelium of the initial segments is composed of ciliated cells and non-ciliated principal cells. The latter are the dominating type and characterized by an apical brush-border and a supranuclear endocytotic apparatus. The stroma of the extratesticular rete contains an abundance of collagen whereas contractile cells are here generally absent. In contrast, the initial segments of the efferent ductules are surrounded by elastic fibres and a layer of contractile cells. All nerves for the feline urogenital junction come from the nervus spermaticus superior. In the epididymal head, small nerve bundles deviate into the septa between the ductules. Single fibres establish a dense network within the muscular coat of the ductuli. At the transition to the extratesticular rete, this network ends abruptly. Nerve fibres in the confines of the rete are associated with blood vessels or proceed to the testicular interior, but establish no relationships with the rete epithelium or the myofibroblasts of the mediastinum. The nervous network in the walls of the efferent ductules and their initial segments is not only composed of sympathetic but also parasympathetic, non-myelinated fibres. Particularly noteworthy is the abundance of calcitonin gene-related peptide (CGRP)- and substance P (SP)-containing axons around the initial segments. Both neuroproteins are consistent markers for sensory neurones. Taken together, it can be assumed that the entry of seminal fluid and spermatozoa into the efferent ductules is controlled by a regulatory nervous chain provided with afferent and efferent components.     

A light and scanning electron microscopic study of the epididymis active state of the endemic Mexican rodent Peromyscus winkelmanni (Carleton) (Rodentia: Muridae)

The macroscopic and microscopic anatomy of the epididymis of the sexually mature Peromyscus winkelmanni (carleton) was examined using light and scanning electron microscopy. The epididymis was divided into three regions: caput, corpus and cauda. The epididymal duct was lined with columnar and cubic epithelium with stereocilia and covered by a muscular connective tissue sheath. Capillaries appear to penetrate directly into the epithelium from the underlying connective tissue in the initial segment. The epididymal epithelium presents four cell types: principal, basal, apical and clear cells. Based on morphological differences (height of epithelial cells, length of the stereocilia, luminal area, larger diameter and spermatic index), the epididymis of P. winkelmanni, presents seven zones. The stereocilia of the epididymal ducts of zones I, II, IV and V are thick and tall, while in zone III they are thin and short. The stereocilia in zone VI are thin, while in zone VII they are short but thick. The secretory products observed in the lumen of the epididymal ducts have vesicular, granular and fibrous form in the seven zones. This study contributes to an understanding of the morphofunctional features of the epididymis in sperm maturation in a species that shows seasonal reproductive activity.     

Effects of age and gonadal steroids on the localization of antigen presenting cells in the epididymis of the male chicken, Gallus domesticus

The goal of this study was to localize antigen presenting cells (APC), which may play roles in defense against pathogens and fertility, and examine the effects of age and gonadal steroids on their population in the rooster epididymis. Healthy White Leghorn male birds (immature 60-day-old birds; matured 150-, 330-, and 550-day-old), and immature birds treated with testosterone propionate (TP) or estradiol benzoate (EB) for 3 or 6 days were used. Cryostat sections of the epididymis and ductus deference were immunostained for Ia to identify APC. RT-PCR was performed to confirm the expression of major histocompatibility complex class II (MHC class II) mRNA in the epididymis. Ia+ cells were localized in the surface epithelium and subepithelial layer of the ductules and occasionally in the luminal content of the epididymis and ductus deference. RT-PCR analysis confirmed expression of MHC class II mRNA in the epididymis, ductus deferens, testis, and spleen. The frequency of Ia+ cells in the subepithelial layer was significantly greater in the proximal efferent ductules than in the other two types of ductules in the epididymis of 550-day-old birds. Although there were no significant differences in the frequencies in the subepithelial layer of the proximal efferent ductules between 60- and 150-day-old birds, the frequencies were significantly greater in 330- and 550-day-old birds than in 60-day-old birds. The frequencies of Ia+ cells in the ductus deferences was increased in the 150-day-old birds compared with the 60-day-old birds, with a larger increase in 330- and 550-day-old birds. The Ia+ cell frequency was significantly increased by EB-injection, but not by TP-injection, on Days 3 and 6 of injection compared with Day 0. These results suggest that the population of APC in the epididymis increases with age after sexual maturation, and estrogen may be one of the factors involved in induction of Ia+ cells.     

Ultrastructural features of the uterus in the sexually immature ostrich (Struthio camelus) during periods of ovarian inactivity and activity

The ultrastructure of the surface epithelium and tubular glands of the uterus in the immature ostrich is described. In ostriches with inactive ovaries the uterus is lined by a non-ciliated simple columnar epithelium, with basally located heterochromatic nuclei. Scanning electron microscopy revealed that these non-ciliated cells have a dense microvillous cover. A simple columnar to pseudostratified columnar epithelium, comprised of non-ciliated and ciliated cells, lines the uterus in birds with active ovaries. The ciliated cells possess a wide luminal region, which contains a nucleus and various organelles. An accumulation of secretory granules was observed in the apical regions of the non-ciliated cells, as well as in a few ciliated cells. In addition to non-ciliated and ciliated cells, a cell type with rarefied cytoplasm was also identified. These cells appear to correspond to calcium secreting cells identified in other avian species. The results of this study indicate that, although uterine differentiation is present in immature ostriches with active ovaries, the production of secretory product appears to occur mainly in non-ciliated epithelial cells.