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.     

An Immunohistochemical Study of the Oviduct in the Domestic Fowl (Gallus domesticus)

This study describes the distribution of vimentin, desmin, smooth muscle actin (SMA) and laminin in the oviduct of the laying domestic fowl. Vimentin immunostaining was localised in the luminal epithelium of the infundibulum, magnum, magnum–isthmus junction and isthmus. The luminal epithelium of the shell gland regions displayed weak vimentin immunostaining. Vimentin immunostaining was demonstrated in the glandular grooves of the tubular infundibular region. In contrast, gland cells in the magnum, isthmus and shell gland regions were vimentin immunonegative. Fibroblasts and vascular endothelial cells in the lamina propria of the oviductal regions studied exhibited vimentin immunostaining. Strong desmin and SMA immunostaining were present in the smooth muscle cells of the tunica muscularis and vascular tunica media. In this study, basement membranes underlying the luminal and glandular epithelia were immunopositive for laminin. In addition, basement membranes associated with smooth muscle cells exhibited laminin immunostaining. The results of the study indicate that the immunolocalisation of desmin, SMA and laminin in the oviduct of the domestic fowl is similar to that in the mammalian uterus. The immunolocalisation of vimentin in the domestic fowl varies depending on the oviductal region.     

Immunohistochemistry with keratin,vimentin, desmin,and α‐smooth muscle actin monoclonal antibodies in canine mammary gland: Benign mammary tumours and duct ectasias

Summary

Duct ectasias (n=2) and different types of benign canine mammary tumours (n=19) were studied immunohistochemically with monoclonal antibodies (MoAbs) directed against various human keratin types (K), α‐smooth muscle actin, vimentin, and desmin. In the duct ectasias and in most tumours the epithelial structures revealed an inner and outer cell layer. The inner cell layer was characterized by labelling with K 7, 8, 18, 19 and mostly also with K 4 and/or K 10 MoAbs. The outer cell layer was almost invariably labelled by K 14, K 14 and 17, and a‐smooth muscle actin MoAbs. The labelling patterns of both duct ectasias and tumours corresponded largely to the patterns observed in normal mammary gland tissue, although a more distinct heterogeneity was seen. Tumours histomorphologically assumed to be of a myoepithelial origin did not show immunohistochemical features of myoepithelial cells. The myoepithelial nature of the vast majority of spindle‐shaped cells present in the adenomas of the complex type and in the fibroadenomas of the benign mixed type could not be confirmed immunohistochemically. These cells, however, unequivocally expressed vimentin, suggesting proliferation of stromal cells in these tumours, which in the fibroadenomas of the benign mixed type may show metaplasia to bone or cartilage.

In the duct ectasias and in some tumours, a fraction of elongated stromal cells, probably representing myofibroblasts, was labelled with the α‐smooth muscle actin MoAb.     

Myofibroblastoma of the neck in a heifer

A 1.5-year-old Holstein heifer had a subcutaneous tumor mass (20 cm diameter) on the ventral portion of the neck, and the tumor was diagnosed as a locally invasive myofibroblastoma. It consisted of moderately cellular fibrous tissue, and the interlobular septum of the thymus was invaded by tumor cells. The neoplastic cells were positive for alpha smooth muscle actin and vimentin, but not for desmin. Electron microscopy disclosed the presence of moderately developed rough endoplasmic reticulum and microfilaments with focal densities.     

Immunohistochemical demonstration of myoid cells in the testis and its excurrent ducts in the domestic fowl

(1) Immunohistochemical methods and three antibodies (against actin, desmin and smooth muscle actin) were used to demonstrate the myoid cells in the domestic fowl testis and its excurrent ducts. (2) A positive reaction to actin, smooth muscle actin and desmin was found in the myoid cells of peritubular tissue of the testis and in rete testis, ductuli efferentes and ductus epididymidis. (3) In the testis myoid-reactive cells form a single layer. In the rete testis, ductuli efferentes and the ductus epididymidis reactive myoid cells form a main component of the stroma. (4) Positive reaction to actin, smooth muscle actin and desmin was also observed in the myoid cells of the tunica albuginea and in the wall of blood vessels in the testis and epididymis, indicating a contractile function for the testicular capsule.     

The expression of intermediate filaments in canine mammary glands and their tumors

Monoclonal antibodies specific for different types of intermediate filaments (cytokeratin, vimentin, desmin and neurofilaments) were used to study the histogenesis of canine mammary glands and 57 canine mammary tumors by immunocytochemistry. The intra- and interlobular duct epithelium, acinar, and intralobular myoepithelial cells stained positively for cytokeratin. Peripheral ductal and acinar cells, as well as interstitial cells, stained positively for vimentin. A similar staining pattern was seen in adenomas, complex adenomas, benign mixed tumors, ductular carcinomas, and one myoepithelioma-like tumor. Additionally, cytokeratin positive cells were scattered interstitially in one single adenoma, most complex adenomas, some benign mixed tumors, complex carcinomas, and in the malignant mixed tumors. All stromal cells stained positively for vimentin. The fibrosarcomas were positive only for vimentin, while the following expressed both desmin and cytokeratin: epithelial-like cells in one adenoma, three complex adenomas, the myoepithelioma-like tumor, the single comedo carcinoma, two complex carcinomas, the single lobular carcinoma, one malignant mixed tumor, and three osteosarcomas. Epithelial-like cells in one adenoma, six complex adenomas, two benign mixed tumors, two complex carcinomas, the lobular carcinoma, and the malignant schwannoma stained for neurofilaments. Three tumors, one adenoma, one complex adenoma, and the lobular carcinoma expressed both desmin and neurofilaments in addition to cytokeratin and vimentin. The results show the expression of different types of intermediate filaments and indicate that there might be a stem cell origin in most of the canine mammary tumors.     

The presence of androgen receptors in the epididymis and prostate of the stallion and cryptorchid horse--a preliminary study

Distribution of androgen receptors (ARs) in the epididymal duct and prostate of three entire stallions and one bilaterally cryptorchid horse was studied immunohistochemically using a polyclonal rabbit antiserum against the ARs. In both the healthy stallions and the cryptorchid, the epithelial cells of the epididymides showed nuclear staining for ARs. The intensity of AR-staining in the principal cells of the epididymis was stronger than that of the basal cells. In the prostate, the glandular secretory cells were moderately stained whereas the basal cells expressed weak AR-staining. Immunostaining for ARs in the reproductive tissues of the cryptorchid horse was always stronger than in those of the stallions. Our results demonstrate for the first time the AR localisation to equine epididymal and prostatic cells, which are directly regulated by androgens.     

Histological and Immunohistochemical Evaluation of Canine Ovary

In the present study, 15 canine ovaries without morphological lesions were examined histologically and immunohistochemically by using a large number of proteins including AE1/AE3, cytokeratin7 (CK7), CK13, CK20, vimentin, desmin, alpha smooth muscle actin (alphaSMA), calponin, S100, Neurofilaments, Inhibinalpha, placental alkaline phosphatase (PLAP) and neuron-specific enolase. Ovarian structures observed in this study included surface epithelium (SE), cortical tubules (CT), tunica albuginea (TA), stromal cells (SC), internal endocrine cells (IE), rete ovarii (RO) and fallopian tubes (FT). SE, CT, RO and FT were broadly immunoreactive for desmin. Besides AE1/AE3 and vimentin, desmin was also closely linked to these structures. Rete ovarii forming a reticular structure showed a positive reaction to S100. Surface epithelium was immunoreactive for PLAP at a significantly high level. In conclusion, these results indicate a specific segment of immunoreactivity as well as the broad range of immunoreactivity in canine ovary. The distinct patterns of immunoreactive for various kinds of proteins will play an important role in facilitating their identification and discrimination even in a normal canine ovary with a complex structure.     

Orbital (retrobulbar) meningioma in a Simmental cow

A 12-year-old Simmental cow was presented with a moderately firm irregular whitish mass of approximately 5 cm in diameter, occupying the right orbit. Microscopically, a poorly differentiated neoplasm was observed. The immunohistochemical panel included cytokeratins, vimentin, epithelial membrane antigen, Factor VIII, CD34, Mart-1, Melan A, smooth muscle actin, desmin, chromogranin, neuron-specific enolase, S-100 protein, and MIB-1. The neoplasm was negative for all of them, with the exception of vimentin and S-100 protein. Transmission electron microscopy revealed abundant desmosomes. These findings support the diagnosis of orbital (retrobulbar) meningioma.     

Histological studies on the regional postnatal differentiation of the epididymis in the ram.

The regional postnatal development of the epididymal duct was studied with histological technique in 22 lambs of the “Swedish Landrace” breed at intervals from 1 week to 18 weeks after birth. At one week, the whole epididymal duct had a simple low columnar epithelium. In the terminal segment the epithelium ecquired a structure similar to that in the adult epididymis at the age of about 6 weeks. Normal spermatozoa were not seen in the cauda epididymidis until the lambs were about 18 weeks old. In the middle segment, distal, intermediate and proximal parts, the epithelium increased gradually in height and reached the adult type at about 12, 15 and 18 weeks, respectively. At about 12 weeks of postnatal life, the epithelium of the initial segment increased rapidly in height, at the same time as a distinct lumen formed in the seminiferous tubules. As far as histological features were concerned, the initial segment showed the adult type when the lambs were about 18 weeks old. The presence of spermatozoa in the cauda epididymidis indicates that the lambs readied sexual maturity at this age. It was concluded that the postnatal differentiation of the ram epididymis starts in the terminal segment and then ascends through the middle and initial segments.     

Congenital hepatic fibrosis in a newborn calf

Congenital hepatic fibrosis was observed in a newborn calf. Light microscopy revealed that periportal areas were linked via connective tissue to the central vein regions and to other periportal areas. Hyperplastic fibers were positive for type I collagen. A remarkable increase in the number of myofibroblasts that were positive for alpha-smooth muscle actin and vimentin was observed in the inner wall of the sinusoids, indicating the occurrence of various fibrogenesis. Ultrastractually, foci of cells resembling cholangiole epithelium cells were observed within the sinusoids, thereby suggesting either ductal plate dysplasia or a bile duct anomaly.     

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.     

Post‐hatch Changes in the Immunoexpression of Desmin,Smooth Muscle Actin and Vimentin in the Testicular Capsule and Interstitial Tissue of the Japanese quail (Coturnix coturnix japonica)

The post‐hatch development of immunoreactivity to desmin, smooth muscle actin (SMA) and vimentin in the testicular capsule and interstitial tissue of day‐old to adult quails was described in this study. The tunica albuginea of the testicular capsule was composed mainly of myoid cells. A zonal arrangement of desmin and SMA immunostaining was observed in myoid cells of the tunica albuginea in 1‐ to 24‐day‐old quails. Immunostaining for SMA and desmin was uniform in the tunica albuginea of adult birds. Vimentin immunostaining in the testicular capsule was demonstrated in mesothelial cells, endothelial cells and fibroblasts. The interstitial tissue contained mesenchymal cells, peritubular myoid cells, Leydig cells and fibroblasts. Desmin‐immunopositive mesenchymal cells were present in the interstitial tissue of 1‐ to 17‐day‐old quails. Peritubular myoid cells expressed strong desmin immunostaining in all developmental stages, while the intensity of SMA immunostaining increased with testicular maturation. Vimentin was demonstrated in Leydig cells and fibroblasts, while the peritubular myoid cells displayed strong vimentin immunostaining only in adult birds. Strong vimentin immunostaining was demonstrated in the endothelial cells of capsular and interstitial blood vessels. The tunica media of these blood vessels displayed desmin and SMA immunostaining. The results of the study have established that variability exists in the distribution and intensity of desmin, SMA and vimentin immunostaining in the testicular capsule and interstitial tissue of the post‐hatch Japanese quail.     

Smooth Muscle Cells of the Bovine Cervical Stroma may have a Secretory, rather than a Contractile Function during Parturition

The bovine cervix contains a large amount of smooth muscle cells distributed over an outer muscular layer and within a stromal layer. The stromal layer exhibits no electromyographic (EMG) activity at parturition. This leads to the question whether the stromal smooth muscle cells of the bovine cervix are prepared to contract with parturition, or whether they have another function. To this end, cervical biopsies were repeatedly taken from 10 pregnant cows at day-185 and -275 of gestation, at spontaneous, uncomplicated calving and at 30 days after calving. The smooth muscle bundles of the stroma were immunohistochemically analysed (n = 5) with regard to their integrity and cellular density, and the degree of staining for connexin-43, smooth muscle actin α (SMA), desmin and vimentin. Additionally, the mRNA expression for connexin-43, SMA, desmin and vimentin was determined with RT-PCR (n = 5). The smooth muscle tissue was arranged in bundles, also at parturition. However, the cellular density of these bundles and the SMA mRNA expression were decreased at parturition. Additionally, the SMA staining and connexin-43 expression and staining remained constant during pregnancy and at parturition. This might indicate that stromal smooth muscle cells are not prepared to contract with parturition, in contrast to the myometrial smooth muscle cells. The smooth muscle cells, stained for SMA, also expressed vimentin, and the proportion of co-expression was increased at day-275 of pregnancy. This suggests that the stromal smooth muscle cells predominantly have a secretory function in cows.     

Effects of ligation of the ductus deferens on the fowl epididymal region

The effects of ligation of the ductus deferens on the epididymis in the fowl were studied histochemically and immunohistochemically to reveal the mechanisms of sperm disposal. At one week post-ligation, the lumina of the rete testes (RT) and the efferent ductules (ED) were distended and filled with densely accumulated spermatozoa. Macrophages and foreign-body giant cells were aggregated in and around the accumulations. The epithelium regressed in the initial portion of the RT with the invasion of fibroblasts and heterophiles into the lumen. The other part of the epithelium was penetrated by many spermatozoa. Numerous lymphocytes and plasma cells infiltrated into the interstitium. At 4 weeks, larger number of spermatozoa agglutinated in the lumen, and large masses of foamy cells and proliferated connective tissue protruded into the lumen. At 8 weeks, large masses of foamy cells were noted. The connecting ductules or the epididymal duct showed no marked changes after ligation. The epithelium of the ED showed weaker or no acid phosphatase activity after ligation. Immunoglobulin G-containing cells increased in number in the interstitium. These results showed that ligation of the ductus deferens in the fowl causes granuloma in the RT and ED, and that epithelial cells, macrophages and granuloma are engaged in the removal of spermatozoa. The participation of antibody is suggested in the sperm disposal processes.     

Feline intraocular tumors may arise from transformation of lens epithelium

Feline ocular sarcomas are malignant intraocular neoplasms that are frequently associated with a history of ocular trauma. They usually present as fibrosarcomas, but some have both epithelial and mesenchymal features. The purpose of this study was to determine the cell of origin of a subset of feline intraocular sarcomas that display a mixed epithelial-mesenchymal phenotype, with elaboration of basement membrane-type matrix. We examined the morphology and histochemical and immunohistochemical phenotypes of nine feline intraocular sarcomas. Immunohistochemistry and in situ hybridization were performed to detect expression of crystallin alpha A. In addition, tumors were examined for expression of vimentin, cytokeratin, smooth muscle actin, desmin, melan A, neural cell adhesion molecule, S-100, glial fibrillary acidic protein, nerve growth factor receptor, and collagen type IV. Animals ranged from 7 to 17 years of age--no breed or sex predilection for tumor occurrence was present. Tumors were characterized by mixed epithelial and mesenchymal phenotypes, both of which elaborated basement membrane-type material and expressed vimentin highly. On the basis of collagen type IV and crystallin alpha A immunopositivity, we established that three of nine tumors were of lens epithelial origin. Expression of desmin and smooth muscle actin identified one tumor as a leiomyosarcoma. The remainder were undifferentiated sarcomas of myofibroblastic origin. This is the first report of lens epithelial neoplasia in clinical material from any species. The history and morphologic features of feline ocular sarcomas are reminiscent of feline vaccine-induced sarcomas. These tumors may share pathophysiologic similarities unique to this species.     

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.     

Immunolocalization of Intermediate Filaments and Laminin in the Oviduct of the Immature and Mature Japanese Quail (Coturnix coturnix japonica)

This study describes the distribution of vimentin, desmin, smooth muscle actin (SMA) and laminin in the oviduct of the immature and mature Japanese quail. The cytoskeletal proteins vimentin, desmin and SMA have been shown to be involved in cellular support, differentiation, migration and contractility. Laminin is a major component of basement membranes. Luminal epithelia in the infundibular and magnal regions of immature and mature birds exhibited strong vimentin immunoreactivity. Luminal epithelial cells exhibiting strong vimentin immunoreactivity were present in the isthmus and shell gland regions of only mature quails. Infundibular glandular grooves displayed strong vimentin immunostaining. In contrast, the glandular epithelia of the magnum, isthmus and shell gland were vimentin immunonegative. Fibroblasts and vascular endothelial cells in the lamina propria of the oviductal regions studied exhibited strong vimentin immunostaining. Smooth muscle cells forming the tunica muscularis and vascular tunica media displayed strong desmin and SMA immunostaining. Strong laminin immunostaining was demonstrated in the basement membranes associated with smooth muscle cells, as well as in the basement membranes underlying the luminal and glandular epithelia. In conclusion, this study has shown that the immunolocalization of desmin, SMA and laminin in the oviduct of the Japanese quail is similar to that in the domestic fowl. However, differences in the immunoexpression of vimentin in the LE of the two avian species were shown to exist. In addition, the study has shown that the immunolocalization of vimentin in the Japanese quail varies depending on the oviductal region, as well as the developmental stage of the oviduct.