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.     

Innervation of the circumvallate papilla of the cat tongue

Immunohistochemically, the distribution of S-100 protein and acetylated tubulin-positive nerve fibres was studied in the circumvallate papilla and its taste buds (TB) in the adult cat. The immunostaining for acetylated tubulin demonstrated an extensive innervation of the circumvallate papilla. Vegetative ganglionic cells were found in the central area of papilla, whereas fine nerve fibres were concentrated under epithelium. Individual positive axons were seen in relation to TB. Nerve fibres enter the TB and branch out among the supporting and sensory cells. Some nerve fibres reach the apical surface of the TB, including the taste pore. Nerve fibres positive for S-100 protein were observed as dense nerve plexus located in the core of the papilla. Satellite cells localized inside the papilla were seen to surround the ganglionic cells. Bands of fine nerve fibres were present under lining epithelium, mainly at the base of the TB. A weak reaction was displayed by taste bud cells and surrounding epithelial cells as well as by the epithelial cells of the papillary side of the moat. A dense network of the nerve fibres was present among the glandular acini and surrounding the ducts of the serous Ebner's glands.     

Structural and immunohistochemical features of the epididymal duct unit of the ostrich (Struthio camelus)

The epididymal duct unit, comprising the ductus conjugens, ductus epididymidis and ductus deferens, was studied histologically, ultrastructurally and immunohistochemically in five sexually mature and active birds. The main morphological features of the pre-dominant non-ciliated (type III) cell of the epithelial lining of this duct unit include, but are not limited to, a moderately abundant smooth or sparsely granulated endoplasmic reticulum, electron-dense secretory granules and numerous mitochondria in the supranuclear zone of the cytoplasm. A single, large heterogeneous lipid droplet, of unknown function, was characteristically situated immediately proximal to the nucleus. The epithelium is obviously secretory and specifically, of the merocrine, and not apocrine, type of secretion. The epithelium of the epididymal duct unit was only focally and weakly to moderately immunopositive to both actin MF and desmin IF, while the duct unit was immunonegative to cytokeratin and vimentin intermediate filaments. The peritubular muscular layer was moderately to strongly positive to both actin and desmin, and negative to cytokeratins and vimentin.     

Histological and histochemical study of the proventriculus (Ventriculus glandularis) of common starling (Sturnus vulgaris)

The histological and histochemical structures of the proventriculus of starling (Sturnus vulgaris) were examined using haematoxylin and eosin and special staining, that is periodic acid schiff (PAS), Masson's trichrome, Alcian blue, Orcein and Reticulin. All three cranial, middle and caudal parts of the proventriculus were also studied. The study results showed that the wall of the proventriculus consisted of mucosal, submucosal, muscular and serosal tunics. The mucosal tunic presented folds and sulci on its luminal surface. In the first third of the proventriculus, the tunica mucosa characterized by presence of folds lined by stratified squamous epithelium and presence of simple tubular glands in the lamina propria. In the middle and caudal thirds of the proventriculus, the surface was covered by a columnar epithelium, and the branched tubular glands were extended through the lamina propria. From the base of the branched tubular glands, the deep proventricular glands were observed that were compound tubuloalveolar lobules. The surface epithelium of the tunica mucosa and the cells lining the proventricular glands showed a positive reaction to PAS and Alcian blue stainings. In addition, the epithelial cells of the tubular and branched tubular glands showed Masson's trichrome-positive reaction. The submucosal tunic was thin in the proventricular wall. The tunica muscularis was formed by a thin inner layer of longitudinal smooth muscle fibres and a thick outer layer of circular fibres. The serosa consisted of loose connective tissue, rich in blood vessels and covered by mesothelium.     

Cytologic,histologic, and immunohistochemical features of maxillofacial alveolar rhabdomyosarcoma in a juvenile dog

Abstract: A 15‐month‐old castrated male dog with a history of intermittent epistaxis and sneezing was admitted for the examination of a maxillofacial mass. An impression smear of a biopsy sample from the cauliflower‐shaped gingival mass contained numerous round cells, 5–25 μm in diameter, which contained a moderate amount of clear to pale blue cytoplasm and resembled lymphoid cells. Mitotic figures were frequently observed. The mass was diagnosed as malignant round cell neoplasia. On histologic examination the tumor was composed of diffusely arranged, small, atypical round cells with a small amount of fibrovascular stroma. Immunohistochemically, the cells were negative for CD3, CD18, CD20, CD79α, cytokeratin, melan‐A, chromogranin A, α‐smooth muscle actin, and myoglobin but positive for vimentin and desmin. The cells also had strong positive nuclear staining for myogenin and MyoD1. A diagnosis of solid‐pattern alveolar rhabdomyosarcoma was made on the basis of morphologic and immunohistochemical results. Alveolar rhabdomyosarcoma should be considered in the differential diagnosis of tumors in juvenile dogs, especially when cytologic findings reveal round, undifferentiated cells.     

Seasonal Changes in the Immunolocalization of Cytoskeletal Proteins and Laminin in the Testis of the Black‐Backed Jackal (Canis mesomelas)

Manipulation of the reproductive activity of jackals is dependent on a thorough understanding of the reproductive biology of this species. This study describes seasonal morphological changes in the adult testis of the black‐backed jackal in relation to the immunoexpression of the basement membrane marker, laminin and the cytoskeletal proteins, cytokeratin, smooth muscle actin and vimentin. Laminin was immunolocalized in basement membranes surrounding seminiferous tubules, as well as in basement membranes associated with Leydig, peritubular myoid and vascular smooth muscle cells. Scalloped basement membranes enclosed seminiferous tubules in regressing testes. The seminiferous epithelium and interstitial tissue in all animals studied were cytokeratin immunonegative. Smooth muscle actin was demonstrated in vascular smooth muscle cells, as well as in peritubular myoid cells encircling seminiferous tubules. Vimentin immunoreactivity was exhibited in the cytoplasm of Sertoli cells, Leydig cells, vascular endothelial cells, vascular smooth muscle cells and fibrocytes. Vimentin immunostaining in Sertoli, Leydig and peritubular myoid cells varied depending on the functional state of the testis. The results of the study have shown that dramatic seasonal histological changes occur in the testes of the jackal. In addition, the use of immunohistochemistry accentuates these morphological changes.     

Histomorphometrical and stereological study of the oesophagus in the adult male Persian squirrel (Sciurus anomalus)

The Persian squirrel (Sciurus anomalus) is habitat in the Middle East countries and feed on pine acorns and other seeds. The present study was carried out to investigate the histological and volumetric features of the oesophagus in Persian squirrels. Five adult male Persian squirrels were included in the study. The cervical, thoracic and abdominal oesophagus of all subjects were processed routinely and sectioned in a serial manner. Then, the total volume of the oesophagus and its different layers were estimated using Cavalieri's principle. Histological assessment revealed a non‐keratinized stratified squamous epithelium lining the mucosa. No glands were seen in the lamina propria and submucosa. Lamina muscularis was present as a distinct layer of smooth muscle cells separating the lamina propria from the tunica submucosa. The tunica muscularis consisted of two distinct layers of striated muscle fibres: inner circular and outer longitudinal that was intermingled with few scattered smooth muscle fibres especially in the abdominal region. The thoracic region contained more amount of the lamina muscularis and tunica muscularis in comparison to the cervical or abdominal regions. The obtained results revealed that the histological structure of the Persian squirrel oesophagus has differences and similarities with other rodents and even with other species of squirrel. These findings would be useful to improve the knowledge in the areas of histological structure of the rodent digestive system.     

Eimeria organisms develop in the epithelial cells of equine small intestine

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.     

Ocular myxoid leiomyosarcoma in a cat

A case of myxoid leiomyosarcoma likely of iris dilator muscle origin in the enucleated eye of a 6-year-old domestic short haired cat is reported. The poorly demarcated mass expanded the iris, partially filled the globe and extended into the optic nerve. The mass was composed of spindle cells separated by abundant matrix positive for mucopolysaccharides with alcian blue. The neoplastic cells were immunoreactive for smooth muscle actin (SMA), S100 and vimentin, and negative for cytokeratin, Melan-A, glial fibrillary protein (GFAP) and desmin. There was no evidence of recurrence or metastasis 6 months after enucleation.     

Ectopic endometrial tissue in mesonephric duct remnants in bitches

Common congenital embryonic remnants of the canine female genital tract are Gartner cysts originating from mesonephric duct remnants. They can increase in size and lead to adverse effects in fertility and health. In the present study, three cases of mesonephric remnants in bitches were analysed. The mesonephric remnants featured an inner lining endometrium comprising surface epithelium, glands and stroma. This ectopic endometrium was further characterized by immunohistochemistry (oestrogen and progesterone receptors, proliferation activity, cytokeratin, alpha smooth muscle actin, and vimentin) and lectin histochemistry compared with normal uterine endometrium. Furthermore, hypertrophic cells at the serosal site of the uteri were detected and analysed in the same way compared with normal serosa. The ectopic endometrium of case no. 2 mesonephric remnant was comparable with normal endometrium whereas in nos 1 and 3 uteri the ectopic endometrium was reduced in thickness. In all mesonephric remnants, surface and glandular epithelial cells of the ectopic endometrium gave positive immunoreactions for cytokeratin, oestrogen and progesterone receptors and showed lectin-binding patterns comparable with normal endometrium. Some of the stromal cells of the ectopic endometria were smooth muscle actin and vimentin positive. Mitotic activity of the ectopic endometria was comparable with normal endometria. Hypertrophic epithelial cells of the serosal side showed positive reactions to anti-oestrogen receptor and anti-cytokeratin immunohistochemistry as well as lectin binding patterns and mitotic activity comparable with the normal canine serosa. The present study is the first considerable immunohistochemical characterization of canine mesonephric remnants and discusses the appearance of ectopic endometrium in mesonephric remnants.     

Endometrial stromal sarcoma with smooth muscle and glandular differentiation of the feline uterus

The intra-abdominal tumor developing in the uterus and lung of a domestic Shorthair cat was examined histopathologically and immunohistochemically. The tumor showed a proliferation of both endometrial stromal and smooth muscle cells accompanied by prominent vasculature. There were well-differentiated endometrial glands, and tubuli made up a monolayer of eosinophilic cuboidal epithelium. Immunohistochemically, the spindle-shaped cells and half of the stromal-like cells reacted to caldesmon and desmin antibodies. The neoplastic epithelium expressed AE1/AE3 cytokeratin. Feline endometrial stromal tumor has, to the best of our knowledge, not been reported previously and has smooth muscle and glandular components that are a unique variant to the human counterpart.     

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.     

Observations on the Right Ovary of Birds of Prey: A Histological and Immunohistochemical Study

In most avian species, only the left ovary and oviduct are developed in the adult bird. Right ovaries and oviducts usually do not mature further after hatching and remain only rudimentary. However, occurrence of a functional right ovary is frequently found in several species of birds of prey. In this study, we investigated the occurrence of the right ovaries and their morphology in these bird species. Four examined wild bird species possessed a right ovary: long‐eared owl, common buzzard, sparrow hawk and goshawk. We used histological and immunohistochemical techniques to evaluate structural differences of the gonads and tried to correlate the findings with folliculogenesis and endocrine functions. The right ovaries showed different sizes and shapes. Cytoskeletal elements (tubulin and vimentin) and α‐smooth muscle actin have been detected in different structures of the right ovaries, but their staining intensity was weaker compared with the left ovary. This shows that also the right ovary is mechanically able to ovulate. We could also demonstrate the expression of oestrogen receptor α and progesterone receptor in the right ovaries, which indicates that also the right ovary can respond to steroid hormone stimuli. We assume that the expression of steroid hormone receptors in the presumptive gonad is still sufficient to mediate the development of a right ovary in the studied species. We conclude that the expression of steroid hormone receptors in the right ovary is involved in its post‐natal development. The histological and immunohistochemical data also imply that in the right ovary, folliculogenesis and ovulation can occur.     

Histomorphology of the stomach, proventriculus and ventriculus of the red jungle fowl

The cranial chamber (proventriculus) and caudal chamber (ventriculus) of the stomach of the Red jungle fowl (Gallus gallus spadiceus) were examined by means of light microscopy. Both chambers presented folds of the tunica mucosa lined by a simple prismatic epithelium that was positive for neutral mucin. Simple tubular glands occupied the lamina propria of both chambers; in the ventriculus of older birds, they showed a coiled base. These ventricular glands were lined by simple cuboidal cells represented by the chief cells and a few large basal cells. The luminal and tubular koilin rodlets and folds of the ventriculus were positive to periodic acid Schiff (PAS) stain. The proventricular glands were situated between the inner and outer layers of the lamina muscularis mucosae. Cells lining the tubulo-alveolar units of the proventricular glands showed a dentate appearance. Vacuoles were not observed, and the cells were negative for Alcian-PAS stain. The tunica submucosa was very thin in the proventricular wall. In the ventriculus, it was not separated from the lamina propria owing to the absence of any lamina muscularis mucosae. The tunica muscularis of the proventriculus was formed by a thick inner layer of circular smooth muscle fibres and a thin outer layer of longitudinal fibres. In addition to these layers, oblique muscle fibres formed the most internal layer of the tunica muscularis in the ventriculus.     

Atypical fibrosarcoma in the skin of a Roborovski hamster (Phodopus roborovskii)

A 2.5‐year‐old intact male Roborovski hamster (Phodopus roborovskii) was presented with a large subcutaneous mass overlying the abdomen, affecting the animal's ambulation and access to different compartments of the cage through narrow tubing. Ultrasound examination delineated a well‐circumscribed mass in the subcutis of the caudoventral abdominal region. The mass was surgically excised and on cytologic examination showed, in a background of blood, a small population of individually arranged oval to spindle‐shaped cells that exhibited a moderate degree of anisokaryosis, coarsely stippled chromatin, one or more prominent nucleoli, and lightly basophilic well‐defined cytoplasmic processes. Histologically, the mass was composed of interlacing streams and bundles of pleomorphic spindle cells (ganglion‐like cells) with variable amounts of collagenous stroma. The neoplastic cells exhibited moderate features of malignancy. These cells stained intensely with vimentin, but not with any other markers, including antibodies to cytokeratin AE1/AE3, S100 protein, desmin, smooth muscle actin, synaptophysin, neurofilament, and androgen receptor. Based on histologic features, the mass was diagnosed as an atypical fibrosarcoma. This is the first report of an atypical fibrosarcoma in a Roborovski hamster and one of few reports of atypical fibrosarcoma in domesticated hamsters overall.     

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.     

Benign Fibrous Histiocytoma of Prepuce in the Mule

Here we present a case of preputial benign fibrous histiocytoma in a 7‐year‐old mule. This case was treated surgically. Pathologic analysis and diagnosis of these lesions were based on a combination of light microscopy and immunohistochemistry. Histologically, the tumour was composed primarily of a storiform pattern of fibroblastic and poligonal cells. In immunohistochemistry, except for consistent immunoreactivity for vimentin, nearly all tumour cells were positive for CD68, S‐100 protein, pan‐cytokeratin, alpha‐smooth muscle actin, and desmin. Benign fibrous histiocytoma of the prepuce in mule is rarely reported in veterinary literature.     

Glomangioma in the Prepuce of a Dog

A case of preputial glomangioma in an 8-year-old Springer Spaniel dog is reported. The dog presented a solitary nodular mass, approximately 3.5 x 5.2 cm in diameter, located on the lateral wall of the prepuce at the base of the penis. Histologically, the excised tumour mass consisted of proliferating round epithelioid cells arranged in a cordon-like structure around small blood vessels. The neoplastic cells were positive for alpha smooth muscle actin and negative for cytokeratin, desmin, S-100 protein and neurone-specific enolase. Based on these pathologic features, the tumour was diagnosed as glomangioma, a neoplasm extremely rare in dogs. To our knowledge, this is the first report of preputial glomangioma in the dog.     

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.     

Immunohistochemical Expression of Nitric Oxide Synthase Enzymes (iNOS,eNOS, nNOS) in the Estrual and Luteal Phases of the Sexual Cycle in the Cow Oviduct

This study was aimed to determine staining intensity, cellular localization and distribution of the nitric oxide synthase (NOS) enzymes during the sexual cycle in the cow oviduct. Oviduct samples belonging to 20 cows, 10 of which were in the estrual phase and 10 in the luteal phase of the sexual cycle, were examined by an immunohistochemical procedure to determine the presence of the NOS enzymes. In the epithelial cells of the isthmus, endothelial NOS (eNOS) expression showed a strong positive reaction during the estrual phase and a weak positive reaction during the luteal phase in the endothelium and smooth muscle of the blood vessels found in the serosa and lamina propria. eNOS expression was not observed in the epithelium of either the ampulla or the fimbria in the two particular phases of the sexual cycle. The eNOS reactions observed in the blood vessel wall in these regions were stronger during the estrual phase. eNOS activity was not observed in the tunica muscularis in any of the regions of the oviduct. During the estrual phase, it was observed that inducible NOS expression showed a stronger positive reaction in the epithelium and muscle layer of the isthmus and ampulla and in the epithelium of the fimbria, compared to the luteal phase. Neuronal NOS immunoreactivity was observed in the epithelial cells of all oviduct regions and in the muscle layer of the isthmus and ampulla and did not display any significant difference between the estrual and luteal phases.