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Topic:Immunohistochemistry
Immunohistochemistry (IHC) is a laboratory technique used to visualize specific antigens in tissue sections of horses through the application of antibodies. This method allows for the detailed examination of the distribution and localization of proteins within equine tissues, providing insights into various physiological and pathological processes. IHC is employed in equine research to study a range of conditions, including infectious diseases, inflammatory responses, and neoplastic disorders. By utilizing specific antibodies that bind to target antigens, researchers can identify cellular and tissue changes, contributing to a better understanding of equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of immunohistochemistry in the field of equine research.
Equine congenital papilloma: pathological findings and results of papillomavirus immunohistochemistry in five cases. Abstract Congenital cutaneous papillomas were identified in five foals. Age at diagnosis ranged from 305 days' gestation to 2 days of age. Breeds were thoroughbred (2), standard-bred (1), warmblood (1) and paint (1). Two were female, one was male and the gender of two cases was not specified. Masses were pedunculated with a roughened, wart-like (verrucous) or a smoother, cauliflower-like surface. Masses occurred as single lesions on the forelimb, lip and face, or as multiple lesions on the trunk. Histological features included relatively uniform epidermal papillary hyperplasia with variable me...
Aquaporin water channels AQP1 and AQP3, are expressed in equine articular chondrocytes. Chondrocytes exist in an unusual and highly variable ionic and osmotic environment in the extracellular matrix of articular cartilage. Alterations to the ionic and osmotic environment of chondrocytes influence the volume and ionic content of the cells, which, in turn, modifies the rate at which extracellular matrix macromolecules are synthesized and degraded. Thus, regulation of the water and solute content of chondrocytes will profoundly affect their anabolic and catabolic functions. The water content of cells is effectively influenced by the abundance of aquaporin (AQP) water channels. Recen...
Investigation of the expression and localization of glucose transporter 4 and fatty acid translocase/CD36 in equine skeletal muscle. To investigate the expression and localization of glucose transporter 4 (GLUT4) and fatty acid translocase (FAT/CD36) in equine skeletal muscle. Methods: Muscle biopsy specimens obtained from 5 healthy Dutch Warmblood horses. Methods: Percutaneous biopsy specimens were obtained from the vastus lateralis, pectoralis descendens, and triceps brachii muscles. Cryosections were stained with combinations of GLUT4 and myosin heavy chain (MHC) specific antibodies or FAT/CD36 and MHC antibodies to assess the fiber specific expression of GLUT4 and FAT/CD36 in equine skeletal muscle via indirect immunofl...
Non-functional C-cell adenoma in aged horses. Thyroid tumours occur in older horses, and most such tumours have been considered to be of follicular epithelial origin. However, their immunohistochemical characterization has not been investigated. The purpose of this study was to confirm a suspicion that most of these tumours are in fact parafollicular cell (C cell)-derived adenomas, and to evaluate their pathogenesis and functional state. Thyroid glands from 38 horses aged 10-29 years were evaluated, all tissue samples being examined histologically, immunohistochemically and ultrastructurally. Nodular tumour masses were found in the thyroi...
The development and distribution of the interstitial cells of Cajal in the intestine of the equine fetus and neonate. This study set out to determine the pattern of development and distribution of the interstitial cells of Cajal (ICC) in the intestinal tract of the equine fetus and neonate. Intestinal tissue samples from 12 naturally aborted equine fetuses and three euthanized neonates were collected and fixed in formalin prior to applying standard immunohistochemical labelling techniques targeting the c-Kit protein of the ICC. At 6 months of gestation, a network of ICC was present in the myenteric plexus region of both the small and the large intestine. ICC were also present within the circular muscle layer....
Endocrine-paracrine cells of the male urogenital apparatus: a comparative histochemical and immunohistochemical study in some domestic ungulates. Specimens of testis, excurrent duct including the male accessory glands and urethra, were studied in boars, bulls, horses and donkeys, in order to localize endocrine/paracrine cells. Silver impregnation methods were used to test the argentaffinity and/or argyrophilia of cells. Immunoreactivities to chromogranin A, 5-hydroxytryptamine, somatostatin, [met]- and [leu]- enkephalins, gastrin-releasing peptide, calcitonin gene-related peptide, neuropeptide Y, substance P, vasoactive intestinal peptide, beta-endorphin antisera were tested by a streptavidin-biotin method. In the testis, epididymis, du...
Use of immunohistochemical marker calretinin in the diagnosis of a diffuse malignant metastatic mesothelioma in an equine. Mesotheliomas are rarely reported in animal species. In this report, the occurrence of a diffuse, metastatic mesothelioma in a 6-year-old gray Arabian mare is described. The mare was presented on clinical examination with ascites, bilateral pleural effusion, and pleural roughening. Necropsy revealed abundant fluid in the abdominal and thoracic cavities. The surface of all organs was thick and fibrosed with multiple raised nodules and hemorrhages. Histology was characteristic of a generalized, biphasic mesothelioma with vascular and lymph nodes metastases. It is believed that the primary tumor ...
Hoof wall wound repair. Surgical stripping of the hoof wall results in a wound that heals remarkabley well. In contrast, lamellae recovering from laminitis are often deformed. Investigating lamellar wound healing may aid understanding of laminitis. Objective: To document temporal changes in the lamellar basement membrane (BM), dermis and epidermis after surgery. Methods: Wall strips were made in the dorsal hoof wall midline of 6 mature horses. Immunohistochemistry was used to document changes in the basement membrane (BM) and detect proliferation of epidermal cells in lamellar tissues harvested at intervals. A confor...
Chronic equine laminitis is characterised by loss of GLUT1, GLUT4 and ENaC positive laminar keratinocytes. Equine laminitis is a multifactorial connective tissue disorder with major implications for the welfare of horses. There are few published studies on phenotypic markers for identification of equine laminar keratinocytes using immunohistochemical techniques. Objective: To establish whether the epithelial sodium channel (ENaC) and the GLUT1 and GLUT4 facilitative glucose transporters may be used as phenotypic markers for identification of equine laminar keratinocytes using immunohistochemical techniques to monitor changes in the keratinocyte population in laminitis. Methods: Histology and immuno...
Desmoplakin and Plakoglobin–specific markers of lymphatic vessels in the skin? Monoclonal antibodies against Desmoplakin and Plakoglobin were tested for their suitability as specific markers of lymphatic vessels. The tissue samples were taken from horse skin in an attempt to establish the horse as a model for human lymphatic diseases. To obtain a clear, positive identification of blood and lymphatic vessels, immunohistochemical staining with antibodies against vascular endothelial growth factor receptor 3 (VEGFR-3) and platelet endothelial adhesion molecule (PECAM-1, CD31), was compared with Desmoplakin and Plakoglobin. Because anti-VEGFR-3 is specific for lymphatic vess...
Assessment of apoptosis in epidermal lamellar cells in clinically normal horses and those with laminitis. To determine and compare the number, type, location, and distribution of apoptotic epidermal cells in the laminae of clinically normal horses and horses with laminitis. Methods: Formalin-fixed samples of digital lamellar tissue from 47 horses (including clinically normal horses [controls; n = 7], horses with acute [4] and chronic [7] naturally acquired laminitis, and horses with black walnut extract-induced [11] or carbohydrate overload-induced [18] laminitis). Methods: Blocks of paraffin-embedded lamellar tissues were stained for DNA fragmentation with the terminal deoxynucleotidyl transferas...
Expression of constitutive endothelial, neuronal and inducible nitric oxide synthase in the testis and epididymis of horse. The expression of three isoforms of nitric oxide synthase (NOS) were examined in the testis and epididymis of a thoroughbred horse. Immunohistochemical studies demonstrated the presence of eNOS immunostaining in some germ cells in the seminiferous tubules and in vascular endothelial cells in the interstitial tissues. Interstitial cells, most likely Leydig cells, were also intensely immunopositive for eNOS. The pattern of immunostaining for nNOS was similar to that for eNOS in the testis. Weak expression of iNOS was detected in the seminiferous tubules of the testis, but intense expression was ...
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 mu...
Gonadotroph heterogeneity, density and distribution, and gonadotroph-lactotroph associations in the pars distalis of the male equine pituitary gland. The intrapituitary mechanisms regulating gonadotrophin secretion in the horse remain unclear. Here, we examined seasonal and gonadal effects on the gonadotroph and lactotroph populations of male horses with the aim of defining a possible morphological basis for the differential release of gonadotrophins. Pituitaries were collected from: (i) gonadal-intact horses in the breeding season (GBS); (ii) orchidectomized horses in the breeding season (OBS); and (iii) orchidectomized horses in the nonbreeding season (ONBS). Immunohistochemistry was performed using antibodies to the luteinizing hormone (...
Immunophenotypic characterization and depletion of pulmonary intravascular macrophages of horses. Pulmonary intravascular macrophages (PIMs) are present in horses and are believed to increase their sensitivity to endotoxin-induced cardio-pulmonary shock. However, owing to a lack of a marker for PIMs and the inability to isolate them, their precise contributions in the horse remain unknown. We designed this study to identify an immuno-phenotypic marker for PIMs and to develop a protocol for their transient depletion with gadolinium chloride (GC). GC is a lanthanide that has been used to deplete liver and lung macrophages. The horses (N = 15) were divided into control (n = 5) and GC-treated ...
Immunohistochemical and functional evidence for a noradrenergic regulation in the horse penile deep dorsal vein. Our aim was to study the presence of noradrenergic nerves and to characterize the alpha-adrenergic receptors involved in the contractions to electrical field stimulation and to alpha-adrenergic agonists of the horse penile deep dorsal vein. Noradrenergic fibres were visualized by immunohistochemistry using an antibody against dopamine-beta-hydroxylase (DBH). For functional studies, the responses of the venous rings to electrical field stimulation and to alpha-adrenergic agonists (noradrenaline, phenylephrine and BHT 920) were studied in the absence and the presence of noradrenergic transmissio...
Physiological and pathological expression of intermediate filaments in the equine endometrium. The aim of this study was to investigate the expression of the intermediate filaments cytokeratin, vimentin and desmin in the equine endometrium by immunohistological techniques. For this purpose, endometrial biopsies of 151 mares were examined to determine physiological cycle patterns and changes resulting from endometriosis. During the physiological cycle epithelial cells and mesenchymal cells express cytokeratin and vimentin, respectively, whilst desmin and vimentin were coexpressed by the smooth muscle cells. Epithelial coexpression of cytokeratin and vimentin was seen in numerous fibrotic...
Cholinergic, nitrergic and peptidergic (Substance P- and CGRP-utilizing) innervation of the horse intestine. A histochemical and immunohistochemical study. The small and large intestine of adult horses were histochemically and immunohistochemically investigated in order to evidence components of the intramural nervous system. The general structural organization of the intramural nervous system was examined by using Nissl-thionin staining as well as the anti-neurofilament 200 (NF200) immunoreaction, which demonstrated the presence of neurons in the submucous as well as myenteric plexuses. The additional presence of subserosal ganglia was shown in the large intestine. Acetylcholinesterase (AChEase) activity was observed in both the submucous and my...
Neuropeptide distribution in the stellate ganglia of the domestic animal. We used immunohistochemical techniques to analyze the localization and distribution of the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) and the neuropeptides methionine-endephalin (M-Enk), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), calretinin (Cal), calcitonin gene-related peptide (CGRP), substance P (SP), and galanin (Gal) in the stellate ganglia of two species of domestic animal (cattle and horses). NPY, VIP and Gal immunoreactive neurons (both cell body and nerve fiber) were observed in the stellate ganglia of both animals. M-Enk and CGRP immunoreactive ne...
Immunohistochemical identification and fiber type specific localization of protein kinase C isoforms in equine skeletal muscle. To investigate whether protein kinase C (PKC) isoforms are expressed in equine skeletal muscle and determine their distribution in various types of fibers by use of immunofluorescence microscopy. Methods: 5 healthy adult Dutch Warmblood horses. Methods: In each horse, 2 biopsy specimens were obtained from the vastus lateralis muscle. Cryosections of equine muscle were stained with PKC isoform (alpha, beta1, beta2, delta, epsilon, or zeta)-specific polyclonal antibodies and examined by use of a fluorescence microscope. Homogenized muscle samples were evaluated via western blot analysis. Results...
Mineralised deposits in the uterine glands of mares with chronic endometrial degeneration. Chronic degenerative disease of the mare's endometrium is characterised by changes in the uterine glands, including cystic dilation, hyperplasia and periglandular fibrosis. Endometrial biopsies were taken from 23 mares with different grades of endometrial degeneration. Solid structures were identified within the lumina of the uterine glands and shown to be calcified by histochemical staining. Most of them were not homogenous but composed of a mixture of mineral and organic substances. Further examinations of these mineralised structures by immunohistochemical methods revealed the presence of t...
Spatial and temporal expression of types I and II receptors for transforming growth factor beta in normal equine skin and dermal wounds. To describe immunolocalization of TGF-beta receptors (RI and RII) in normal equine skin and in thoracic or limb wounds, healing normally or with exuberant granulation tissue (EGT). Methods: Group A: six wounds on one metacarpus and one midthoracic area. Group B: six wounds on both metacarpi, one of which was bandaged to stimulate EGT. Immunohistochemistry was used to detect RI and RII expression in wound margins. Methods: Eight horses, randomly assigned to one of two study groups. Methods: Neutralizing polyclonal anti-rabbit RI and RII antibodies were used to detect spatial expression of RI an...
Immunocytochemical localization of cytochrome P450 aromatase in the testis of prepubertal, pubertal, and postpubertal horses. The large amount of testicular estrogens produced by the stallion is unique compared to the amounts found in other domestic species. Although the cellular locale of the cytochrome P450 aromatase (P450arom) enzyme that converts C19 androgens to C18 estrogens has been identified in the Leydig cell of adult equine testis, the location in the immature equine testis is not known. The goal of this work was to localize the enzyme in colts and stallions during sexual development. Testes were obtained from prepubertal (n=7), pubertal (n=6), and postpubertal (n=8) colts and stallions during both the bre...
The equine thymus microenvironment: a morphological and immunohistochemical analysis. We characterized herein the microarchitecture of the equine thymus along with post-natal development (6 months-->18 years). Thymuses showed an involutional process, beginning before the puberty and defined by five histological grades, which consider the progressive cortical thymocyte depletion, shrinkage and rearrangement of the epithelial network and increase in extracellular matrix (ECM). A second feature of the equine thymus was the presence of eosinopoiesis, erythropoiesis, mastocytopoiesis and plasmacytogenesis. Additionally, lymphatic vessels, full of lymphocytes, were particularly pr...
Connective tissue growth factor in tear film of the horse: detection, identification and origin. Healing of corneal ulcers in horses is often associated with profound corneal stromal fibrosis and scar formation resulting in visual impairment. Connective tissue growth factor (CTGF) is a fibrogenic cytokine involved in wound healing and scarring. The purpose of this study was to determine whether CTGF was present in the tear fluid of normal horse eyes and the eyes of horses with corneal ulcers in order to evaluate the role of CTGF in corneal wound healing and corneal scar formation. Methods: Tear fluid samples were collected from 65 eyes of 44 horses; 32 samples from normal eyes, 21 samples...
Equine viral arteritis in a newborn foal: parallel detection of the virus by immunohistochemistry, polymerase chain reaction and virus isolation. A 4-days-old foal died after a short course of respiratory syndrome and fever. Large areas of the alveoli, bronchioles and bronchi were partly or completely filled by hyaline membranes. Pronounced oedema and mild interstitial pneumonia were present and, in the small muscular arteries, fibrinoid necrosis and vasculitis or perivasculitis could be seen. Vasculitis was found in several other organs, and it was most severe in the thymus. The virus was detected in the lung, kidney and spleen using virus isolation and in the lung and spleen using polymerase chain reaction. The virus was also detected...
Screening for West Nile virus infections of susceptible animal species in Austria. Avian mortality and encephalomyelitis in equines are considered good indicators for West Nile virus (WNV) activity. We retrospectively tested 385 horse sera for WNV antibodies and looked for WNV nucleic acid and/or WNV antigen in paraffin embedded tissues from 12 horses with aetiologically unresolved encephalomyelitis and 102 free-living birds of different species which had been found dead. With the exception of four horses originating from eastern European countries investigated on the occasion of transit through Austria, all horse sera were negative. Nested RT-PCR of the horse tissues yielde...
Experimental analysis of error sources in fibre type counts of biopsies in horses. The contribution to total variance of different error sources in fibre type counts of equine gluteus medius muscle biopsies was determined to quantify and possibly improve the resolution of the method. Fibre types were defined on the basis of myosin heavy chain immunostaining. Errors were determined at levels: (1) positioning the insertion channel, (2) positioning the needle tip (3) biopsy heterogeneity (4) observer interpretation. Errors at levels 1 and 2 were considerable. Confidence intervals for individual observations were +/- 10-15%. In longitudinal studies a group size of 4 animals is n...
Expression of key prostaglandin synthases in equine endometrium during late diestrus and early pregnancy. Luteolysis in domestic species is mediated by the release of luteolytic pulses of prostaglandin (PG) F(2alpha) by the uterus at the end of diestrus, which must be suppressed by the conceptus to permit maternal recognition of pregnancy. In many species, including the horse, both the conceptus and the endometrium also synthesize PGE(2), which may antagonize PGF(2alpha) by playing a luteotropic and/or antiluteolytic role. While the release of PGE(2) and PGF(2alpha) by the equine endometrium in late diestrus and early pregnancy has been previously studied, the underlying prostaglandin synthase gen...
Multicentric leiomyosarcoma in a young male horse. A 2-year-old male Thoroughbred was evaluated for multiple firm, raised nodules of the maxilla and mandible that had been present since 6 months of age. The horse was evaluated as a yearling with a final histologic diagnosis of fibrosarcoma. As a 2-year-old, the colt made a loud upper airway sound and was agitated after strenuous exercise. Results of upper airway endoscopy indicated severe narrowing of the nasal passages and pharynx with normal resting function. Radiography revealed multiple circular bony lesions in the mandible, maxilla, multiple long bones, and cervical vertebrae. Biopsy spec...
