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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.
[A case of cutaneous epitheliotropic malignant lymphoma (mycosis fungoides) in a horse]. This article describes an eleven year old mare with apathy, fever, enlarged mandibular lymph nodes, skin lesions on the upper lip and edematous, grey-red mucous membranes in the nose, mouth and vulva. Histopathology revealed infiltrates with atypical lymphocytes forming Pautrier's microabscesses. The neoplastic cells had large, often indented nuclei. Immunohistology showed that some cells were CD3-positive (Pan T-cell-marker). The diagnosis of cutaneous epitheliotropic malignant lymphoma (Mycosis fungoides) was made. The etiology in the horse is unknown.
Sensory epithelium of the vomeronasal organ express TrkA-like and epidermal growth factor receptor in adulthood. An immunohistochemical study in the horse. The medial wall of the vomeronasal organ (VNO) is lined with a sensory epithelium that is closely related to the olfactory epithelium, which is developed from the olfactory placode. It undergoes continuous replacement during its life span. In other sensory epithelia, cell proliferation is under the control of some trophic factors. Whether these proteins are involved in the continuous turnover of the VNO epithelium is unknown. This study approaches this topic by analyzing the occurrence of signal-transducing receptor proteins for neurotrophins (Trk proteins) and epidermal growth factor (EGFr). ...
Orbital neuroendocrine tumors in three horses. Three horses were examined because of exophthalmos and others signs indicative of a space-occupying orbital mass. In 2 horses, exenteration was used to remove the orbital mass. In a third horse, an orbital tumor and lung metastases were found at necropsy. Routine histologic and Grimelius' histochemical staining were used on fixed tissues; immunohistochemical staining for chromogranin and electron microscopy also were performed. Masses were orbital neuroendocrine tumors. Horses that underwent exenteration were alive 19 and 24 months after surgery. Hypotension was encountered at surgery in both ...
Fatal encephalitis due to novel paramyxovirus transmitted from horses. In September, 1994, an outbreak of severe respiratory disease affected 18 horses, their trainer, and a stablehand in Queensland, Australia. Fourteen horses and one human being died. A novel virus was isolated from those affected and named equine morbillivirus (EMV). We report a case of encephalitis caused by this virus. Results: A 35-year-old man from Queensland had a brief aseptic meningitic illness in August, 1994, shortly after caring for two horses that died from EMV infection and then assisting at their necropsies. He then suffered severe encephalitis 13 months later, characterised by unc...
Biochemical and site-specific effects of insulin-like growth factor I on intrinsic tenocyte activity in equine flexor tendons. To examine the site-specific and dose-dependent effects of insulin-like growth factor I (IGF-I) on normal equine tendon in vitro. Methods: Superficial digital flexor tendon explants derived from a euthanatized 3-year-old horse. Methods: Explants in culture were treated with 0, 100, 250, or 500 ng of IGF-I/ml for 14 days with an end-stage radiolabel of 20 microCi of [3H]proline/ml or 5 microCi of [3H]thymidine/ml. The tendon tissues were then analyzed biochemically for hydroxyproline content by reverse-phase high-performance liquid chromatography, DNA content by fluorometry, and glycosaminoglyc...
The equine endometrial mast cell during the puerperal period: evaluation of mast cell numbers and types in comparison to other inflammatory changes. Endometrial biopsies of 44 broodmares were histologically examined on days 3, 6, and 9 postpartum. The mares were subdivided into three groups according to the course of the puerperal period. In 29 mares, parturition and expulsion of the placenta was normal, six mares showed dystocia, and in nine mares, the placenta was retained for > 2 hours. Tissue samples were evaluated histologically, and the average numbers of granulocytes, lymphocytes, macrophages, siderophages, and mast cells was determined. Protease content of mast cells was examined with a double-enzyme immunohistochemical staining...
Immunohistochemical demonstration of equine herpesvirus-1 antigen in neurons and astrocytes of horses with acute paralysis. Equine herpesvirus-1 (EHV-1) infection in a few widely scattered neurons and astrocytes plus endothelial cells in brain and spinal cord of two horses with naturally occurring paralytic disease was demonstrated by use of an immunoperoxidase technique. These horses were euthanatized less than 48 hours after the onset of clinical signs. No staining for EHV-1 was demonstrated in brain or spinal cord of three horses that had a longer duration of clinical disease or in two uninfected horses.
Biochemical, histochemical, and immunohistochemical characterization of distal tibial osteochondrosis in horses. To compare the biochemical, histochemical, and immunohistochemical profiles of articular cartilage from horses with naturally acquired distal tibial osteochondrosis (OC) with cartilage from a similar location in clinically normal horses. Methods: 9 affected horses (group 1, 16 OC lesions) and 4 control horses (group 2, 8 normal osteochondral specimens). Methods: OC specimens were collected during arthroscopic removal of the fragment, and control specimens were collected by aseptic osteotomy. Uronic acid, total protein, total glycosaminoglycan (GAG), chondroitin sulfate (CS), and keratan sulfat...
Proteoglycan metabolism of equine articular chondrocytes cultured in alginate beads. Equine chondrocytes were cultured in vitro for 30 days in ionically gelled alginate beads. The alginate polymerises into a stable gel in the presence of divalent cations (calcium), and rapid depolymerisation in the presence of a calcium chelator releases the viable chondrocytes. The chondrocytes maintained a spherical appearance for 30 days in culture, in marked contrast to monolayer cultures, which develop a dedifferentiated fibroblastic morphology. The major proteoglycan molecule produced by the encapsulated chondrocytes was aggrecan, of similar hydrodynamic size to aggrecan molecules presen...
Neosporosis as a cause of equine protozoal myeloencephalitis. Neosporosis was diagnosed in an 11-year-old Quarter Horse gelding with clinical signs and diagnostic test results compatible with equine protozoal myeloencephalitis (EPM). Presumptive postmortem diagnosis of EPM attributable to Sarcocystis neurona infection is generally made on the basis of detecting an antibody titer to S neurona in the CSF or characteristic histologic lesions, even when parasites have not been specifically identified. Neosporosis was confirmed in the horse described here by use of immunohistochemical examination, in vitro culturing, and ultrastructural and molecular characte...
[Immunopathology of Borna disease in the horse: clinical, virological and neuropathologic findings]. Tissues from nine horses and one donkey suffering from natural Borna disease were investigated. Clinically, all animals demonstrated progressive reduced mentation and aggravating gait disturbances. During the clinical course anorexia and progressive loss of proprioception were observed. Cranial nerve failure was accompanied by signs of pharyngeal paralysis, sialorrhea, bruxism, and by blindness. Virologically, infectious virus was detected in the brain of all animals investigated but was not found regularly in all areas of the brain. However, in all cases, infectivity was found in the thalamus...
Immunohistochemical examination of light-chain expression (lambda/kappa ratio) in canine, feline, equine, bovine and porcine plasma cells. Five species of domestic animals were examined immunohistochemically and the light-chain ratios of the immunoglobulins produced by plasma cells analysed. Forty dogs, 11 cats, 10 horses, 11 cattle and 14 pigs were tested using the sequential indirect immunoperoxidase and immunophosphatase methods. Tissues from the tonsils, spleen and cervical lymph nodes were analysed. It could be seen that the lambda/kappa ratio in dogs, cats, horses and cattle is largely dominated by the lambda chains (lambda/kappa ratio in dogs: 91/9%, in cats; 92/8%; in horses: 96/4%; in cattle: 91/9%). A more or less balan...
Chronic bronchopulmonary aspergillosis in a horse with Cushing’s syndrome. An atypical case of chronic equine bronchopulmonary aspergillosis with an unusual hyphal morphology was diagnosed in a horse with Cushing's syndrome. Because of the hyphal localization in chronic ectatic bronchi and bronchioles, and juxtabronchiolar processes, the observed type of aspergillosis is similar to 'saprophytic bronchopulmonary aspergillosis' or 'semi-invasive pulmonary aspergillosis' in humans. The aetiological diagnosis of aspergillosis was accomplished by the application of a panel of monospecific polyclonal and monoclonal antibodies in immunohistochemical techniques.
Cutaneous leishmaniasis in two horses. This report contains the clinical, histologic, immunohistochemical, and electron microscopic findings in two cases of equine cutaneous leishmaniasis. Nodular, sometimes crusty or ulcerated lesions were confined to the pinna and adjacent neck in both animals. The dermal inflammatory infiltrate was lymphohistiocytic in horse No. 1 and pyogranulomatous with formation of tuberculoid granulomas in horse No. 2. Numerous Leishmania organisms were found within macrophages in both animals. There was moderate to intense and specific reaction by immunoperoxidase using a polyclonal antiserum against Leish...
Immunolocalization of aromatase P-450 in ovarian tissue from pregnant and nonpregnant mares and in ovarian tumours. Aromatase P-450 (P-450arom) is a crucial regulatory enzyme that is necessary for conversion of androgens to oestrogens. Corpora lutea and follicles were obtained from the ovaries of cyclic mares and from mares at day 20 and days 40-70 of pregnancy. The presence of P-450arom within specific cell types was investigated by immunostaining to determine potential sites of oestrogen synthesis. Immunoreactivity for P-450arom was confined to the granulosa layer of non-atretic follicles > 5 mm in diameter and to corpora lutea at all stages of the oestrous cycle and during pregnancy. These findings co...
Multiple peripheral nerve sheath tumors in the small intestine of a horse. Multiple neurofibromas, schwannomas, and hyperplastic enteric plexuses were observed in the distal jejunum and ileum of a 6-year-old pinto gelding. The animal was presented because of an acute episode of colic. Three meters of distal small intestine, partially incarcerated in the epiploic foramen, were surgically removed. Numerous tumor nodules up to 10 mm in diameter were found adjacent to a Meckel's diverticulum, predominantly located in the subserosa of a hypertrophic segment. Histologically, tumors were well demarcated and composed of interlacing fascicles formed by spindloid cells. Adjace...
Correlation between myofibrillar ATPase activity and myosin heavy chain composition in equine skeletal muscle and the influence of training. The histochemical myofibrillar ATPase (mATPase) method is used routinely for identification of equine skeletal muscle fiber types, but important problems have been observed with the subdivision of fast fiber population when using this method. To verify the use of this qualitative method, a number of equine muscle biopsies were analyzed with a combination of histochemical, immunohistochemical, electrophoretic, and morphometric techniques. The influence of training on these interrelations was also evaluated. Methods: Five young (2-3 years old) thoroughbred horses were intensively trained for 8 m...
Myosin heavy chain isoforms in adult equine skeletal muscle: an immunohistochemical and electrophoretic study. The aim of this study was to characterize the myosin heavy chain (MyHC) isoforms present in equine skeletal muscle. Methods: Muscle biopsies were removed from the superficial region of the gluteus medius muscle of five mature horses and analyzed by immunohistochemistry (using a battery of monoclonal antibodies specific for rat MyHC isoforms) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results: Immunohistochemistry allowed subdivision of three different muscle fiber populations containing a single MyHC, one slow and two fast, and two hybrid populations, one containing slow an...
Immunohistochemical analysis of an equine model of synovitis-induced arthritis. To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone. Methods: 2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2). Methods: Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 micrograms/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and a morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articula...
Papillary ductal adenocarcinoma in the mammary glands of two horses. Mammary gland neoplasms in horses are uncommon and may initially be confused with mastitis. Masses from the mammary glands of two horses with chronic discharges were examined by cytology and histopathology. Both masses were diagnosed as papillary ductal adenocarcinomas with extensive intraductal and intralobular involvement and only focal infiltration of the adjacent stroma. Complementary immunohistochemical studies were made of both fresh and formalin-fixed tumour tissue, and attempts were made to assess the steroid receptor status of fresh tissue biochemically to assess the possibility of ho...
Ubiquitin expression in degenerating axons of equine cervical compressive myelopathy. Neuropathologic examination revealed axonal swelling and breakdown leading to Wallerian degeneration of affected myelinated nerve fibers in the spinal cord white matter of four young horses with equine cervical compressive myelopathy. Immunohistochemical reactions for the cell stress protein ubiquitin revealed an enhanced presence in the swollen axons, which may reflect a role for ubiquitin in the neuronal catabolic process of axonal compression and degeneration in this myelopathy.
Immunohistochemical detection of virulence-associated antigens of Rhodococcus equi in pulmonary lesions of foals. Rhodococcus equi was isolated from the lungs of six foals with bronchopneumonia. All isolates expressed 15-17-kd antigens by immunoblot analysis and contained a virulence-associated plasmid of 85 or 90 kb. Immunohistochemically, R. equi from all pulmonary lesions showed the expression of 15-17-kd antigens mainly in the phagocytic cells. The specific monoclonal antibody to 15-17-kd antigens of R. equi (MAb 10G5) may be an aid in the diagnosis of R. equi-induced pneumonia.
Expression of types II, VI and X collagen in equine growth cartilage during development. The synthesis and expression of collagen types II, VI and X were investigated in growth cartilage selected from a group of 31 horses and ponies in the age range 157 days of gestation to 12 years. Collagen isolation, immunolocalisation and in situ hybridisation techniques were used in order to provide information on the pattern of synthesis of these 3 collagens during endochondral ossification in normal horses. Type II collagen immunoreactivity and mRNA expression was found in each of the 3 zones of growth cartilage chondrocytes in all samples studied, whereas the localisation of both collagen ...
Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA-III) in the reproductive tract of male horses. To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA-I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. Methods: Descriptive and immunohistochemical study. Methods: 4 clinically normal male horses. Methods: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimen...
Myosin isoforms and muscle fiber characteristics in equine gluteus medius muscle. To date, four different myosin heavy chain (MyHC) isoforms have been identified in adult skeletal muscle of a number of species: types I, IIa, IIx or IId, and IIb. The aim of this study was to investigate the distribution of various MyHC isoforms in the equine gluteus medius and gluteus profundus muscles in relation with several morphometric variables of muscle fibers. Methods: Samples from different depths of the gluteus medius muscle (2, 4, 6, and 8 cm) and gluteus profundus muscle of five sedentary horses were examined by MyHC gel electrophoresis, monoclonal antibodies staining against fast...
Histopathological and immunofluorescent studies on transplacental infection in experimentally induced abortion by equine arteritis virus. Five pregnant mares, at between 6 and 8 months gestation, were experimentally infected with the Bucyrus strain of equine arteritis virus (EAV). Of the five mares, four aborted and one died. The pathogenesis of the abortions was studied, using histopathologic techniques, tissue immunofluorescence and virus isolation. Common microscopic lesions in the maternal reproductive organs indicated myometritis with a degeneration of the myocytes and an infiltration of the mononuclear cells. Epithelial cells of the endometrial gland showed sporadic degeneration. Lesions in the fetal tissue included an atr...
Immunohistochemical diagnosis of eastern equine encephalomyelitis. An immunohistochemical (IHC) assay was developed for the detection of eastern equine encephalomyelitis (EEE) virus antigen in formalin-fixed, paraffin-embedded tissues. All cases of EEE diagnosed at the Michigan State University Animal Health Diagnostic Laboratory from 1991 through 1994 were evaluated. The diagnosis was based on histopathologic examination of the brain and confirmatory virus, isolation. Sections of cerebrum from 26 equids and 5 birds were assessed by IHC. Histologically normal brain tissues from 2 horses and 1 pheasant and brain tissues from 2 cases of equine neurologic diseas...
Histologic and immunohistochemical characterization of hemangiomas in the skin of seven young horses. The histologic and immunochemical characteristics of benign vascular tumors excised from the skin of seven young horses were evaluated. The patients were male horses of various breeds and were 16 months of age or younger at the time of presentation. Six tumors occurred on the extremities, and one was removed from the lip. Histologically, most tumors consisted of cellular nodules of varying compactness with few to many blood-filled lumina. Nodular growth separated preexisting adnexa and subcutaneous collagen. Some tumors contained or consisted predominantly of loosely packed arrays of ramifying...
