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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.
Inhibin secretion in the mare: localization of inhibin alpha, betaA, and betaB subunits in the ovary. To determine the source of circulating inhibin and estradiol-17beta during the estrous cycle in mares, the cellular localization of the inhibin alpha, betaA, and betaB subunits and aromatase in the ovary was determined by immunohistochemistry. Concentrations of immunoreactive (ir-) inhibin, estradiol-17beta, progesterone, LH, and FSH in peripheral blood were also measured during the estrous cycle in mares. Immunohistochemically, inhibin alpha subunits were localized in the granulosa cells of small and large follicles and in the theca interna cells of large follicles, whereas inhibin betaA and ...
Immunohistochemical studies in equine recurrent uveitis (ERU). Despite extensive clinical research, the etiology of equine recurrent uveitis (ERU) is still unknown. After an immunologic pathogenesis was established in recurrent uveitis in humans, a similar pathogenic mechanism was assumed to exist in ERU. To investigate whether immunopathologic mechanisms are involved in ERU, 20 eyes of 15 horses with ERU were examined immunohistochemically with a T cell marker, B cell marker, and anti-major histocompatibility complex (MHC) class II antibodies. Twenty-six eyes of 20 horses were used for investigation of MHC class II antigen expression in normal equine eye...
Immunohistochemical localization of the spermadhesin AWN-1 in the equine male genital tract. Spermadhesins are proteins with various functions in sperm capacitation and zona pellucida binding. In this study the cellular localization of the spermadhesin AWN-1 has been examined in the equine male genital tract. Results obtained by immunohistochemical methods reveal that in the horse AWN-1 is synthesized in spermatogonia, in the rete testis, the ductus epididymidis and the seminal vesicles. These findings indicate that the cellular origin of spermadhesins is species-specific.
Immunohistochemical demonstration of African horse sickness viral antigen in tissues of experimentally infected equines. African horse sickness virus (AHSV) antigen was demonstrated immunohistochemically in formalin-fixed, paraffin-embedded sections of tissues collected from three ponies suffering from the peracute form of the disease and from one pony affected by the fever form. The pattern of the antigen distribution indicated a particular organ tropism characterised by an accumulation of AHSV antigen in cardio-pulmonary tissues of the animals with the peracute disease and in the spleen of the pony with the fever form. AHSV antigen was identified in endothelial cells of small blood vessels, particularly capill...
Expression of inhibin alpha-subunit in horse testis. Inhibin is believed to play roles in the pituitary secretion of FSH and in the paracrine regulation of testicular function. Although it has been generally accepted that inhibin is produced in Sertoli cells, there was a recent evidence for the localization of inhibin in Leydig cells of primates, rat and sheep. However, there is no report on the expression of inhibin in the adult horse testis. Therefore, using immunohistochemistry, western blotting and in situ hybridization techniques, the present study examined inhibin alpha-subunit (Ih-alpha) expression in the adult horse testis. For the detec...
Progesterone-induced acrosome reaction in stallion spermatozoa is mediated by a plasma membrane progesterone receptor. The aim of the present study was to investigate whether the induction of stallion sperm acrosome reaction (AR) by progesterone is mediated by binding of progesterone to a receptor on the sperm plasma membrane or to an intracellular progesterone receptor. Progesterone-BSA conjugate labeled with fluorescein isothiocyanate (P-BSA-FITC) in combination with a vital stain, ethidium homodimer, was applied to visualize the presence of the progesterone receptor on living spermatozoa. Alternatively, an indirect immunofluorescence technique employing a monoclonal antibody (C-262) against human intracellu...
Transfer of a uterine lipocalin from the endometrium of the mare to the developing equine conceptus. One of the major, progesterone-dependent proteins secreted into the uterine lumen of the mare is a 19-kDa lipocalin (P19). It associates strongly with the embryonic capsule that envelops the young horse conceptus in early gestation, suggesting that it may be involved in sustaining early development. However, it was not known whether the protein was transported through the capsule and/or trophoblast layer and into the yolk sac cavity. To address this question, polyclonal antisera were raised against a C-terminal peptide (based on the deduced amino acid sequence of P19) and a recombinant-derived...
Functional anatomy of the cartilage of the distal phalanx and digital cushion in the equine foot and a hemodynamic flow hypothesis of energy dissipation. To examine macro- and microscopic characteristics of cartilage of the distal phalanx (ungual cartilage [UC]) and digital cushion in the equine foot and to relate them to the foot's function of energy dissipation. Methods: 85 horses and 5 foals of various breeds and ages. Methods: Feet, obtained at necropsy, were perfused with India ink (n = 30), latex (5), or polymer plastic (10). Select feet were examined histologically for tissue architecture and to identify elastic fibers. Immunochemistry to identify substance P peptides in nerves (feet from foals) and gold chloride impregnation of axons (n...
An immunohistochemical investigation of the adult stage of the equine parasite Strongylus vulgaris. Adult Strongylus vulgaris, collected from the caecum of infected horses and embedded in paraplast using standard methods, were sectioned for immunohistochemistry (IHC) studies. Antibodies were raised in rabbit against the excretory-secretory product (ESP) and against two constituent protein bands (28-30 kDa). The use of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting indicated the immunogenicity of ESP and of the subunits (28-30 kDa). In ELISA, both rabbit hyperimmune sera recognized the ESP and (28-30 kDa) ban...
Dysgerminoma in an Arabian filly. A yearling Arabian filly was referred to the Veterinary Medical Teaching Hospital with a history of weight loss, profound anemia, and peritoneal effusion. At necropsy, a large, soft, mottled tan and red neoplastic mass was at the pelvic inlet replacing the left ovary. Additional tumor nodules of various sizes were disseminated throughout the mesentery, diaphragm, and serosal surfaces of the abdominal viscera. Histologically, the neoplasm had sheets of large round to polygonal cells separated into lobules by fibrous connective tissue with multifocal areas of necrosis. Tumor cells stained strong...
Equine malignant lymphomas: morphologic and immunohistochemical classification. Gross lesions, microscopic appearance, and immunophenotyping are reported in a retrospective study of 31 cases of equine malignant lymphoma. Immunohistochemical studies were performed on archived formalin-fixed, paraffin-embedded tissues. Monoclonal antibodies to surface glycoprotein BLA.36 and intracytoplasmic domains of mb-1 and B29 were used to document the presence of B lymphocytes in the equine tumors. Polyclonal antibody to CD3 and monoclonal antibodies to T-lymphocyte markers CD3 and CD5 revealed the presence of variable numbers of T cells within the equine lymphomas. The neoplastic com...
Testicular inhibin in the stallion: cellular source and seasonal changes in its secretion. The cellular localization of inhibin alpha, betaA, and betaB subunits, 3beta-hydroxysteroid dehydrogenase (3beta-HSD), and cytochrome P450 aromatase (aromatase) in stallion testes was investigated. In addition, detailed seasonal changes in circulating immunoreactive (ir)-inhibin were investigated in correlation with testosterone, estradiol, LH, and FSH. Inhibin alpha subunit-positive staining was observed in Sertoli cells, and more clearly positive staining was noted in Leydig cells. Inhibin betaA and betaB subunits were also stained in both types of cells. Immunoreactivity of 3beta-HSD and ar...
Monoclonal antibody against Babesia equi: characterization and potential application of antigen for serodiagnosis. Monoclonal antibody (MAb) BEG3 was produced against Babesia equi parasites to define a species-specific antigen for diagnostic use. The MAb reacted with single, paired, and Maltese cross forms of B. equi, and no reaction was observed with this MAb on acetone-fixed Babesia caballi, Babesia ovata, or Babesia microti parasites in the indirect immunofluorescent antibody test. Confocal laser and immunoelectron microscopic studies showed that the antigen which was recognized by this MAb was located on the surface of B. equi parasites. This MAb recognized a 19-kDa protein of B. equi antigen and did n...
Objectivity of two methods of differentiating fibre types and repeatability of measurements by application of the TEMA image analysis system. The objectivity of two of the most widely used methods for differentiation of fibre types, i.e. 1) the myosin ATP-ase method (Brooke and Kaiser, 1970a,b) and 2) the combined method, by which the myosin ATP-ase reaction is used to differentiate between fast and slow twitch fibres and NADH-tetrazolium reductase activity is used to identify the subgroups of fast twitch fibres (Ashmore and Doerr, 1970, Peter et al., 1972), was assessed in muscle samples from horses, calves and pigs. We also assessed the objectivity of the alpha-amylase-PAS preparation for the visualisation of capillaries (Andersen...
Spontaneous Borna disease in sheep and horses: immunophenotyping of inflammatory cells and detection of MHC-I and MHC-II antigen expression in Borna encephalitis lesions. Borna disease (BD) has been recognized as a virally induced T-cell dependent immunopathological disorder of the central nervous system (CNS), as shown by experimental infection of rats with Borna disease virus (BDV). In contrast to the rat model, little is known about the pathogenesis of spontaneous BD in sheep and horses. The present study describes the brain lesions of 12 ovine and 11 equine cases of naturally occurring BD. A set of monoclonal and polyclonal antibodies was used in order to determine the cells operative in encephalitic lesions and to detect expression of MHC-I and MHC-II prod...
C-cell adenoma containing variously sized thyroid follicles in a horse. A thyroid gland tumor, showing unusual histology, was identified in a 13-year-old male Andalusian horse. Microscopically, the tumor consisted of neoplastic proliferation of C-cell (parafollicular cell) with cytoplasmic fine granules, containing diffusely distributed, variously sized colloid-containing follicles. Immunohistochemically, the neoplastic C-cell were positive for calcitonin and follicleforming epithelial cells showed a positive reaction for thyroglobulin. Ultrastructurally, membrane-bound secretory granules up to 250 nm in diameter were found in the cytoplasm of the parafollicular c...
Cutaneous, multilocular T-cell lymphosarcoma in a horse–clinical, ultrasonographic and pathological findings. Clinical, ultrasonographic and pathological findings of a cutaneous, multilocular T-cell lymphosarcoma (malignant lymphoma) in a 13-year-old, brown Austrian warmblood mare are reported. The horse was under clinical observation, and the tumours were sonographically monitored over an 8 month period, revealing worsening of body condition, weight loss, lameness of the left hindlimb and a slight increase in the tumours size. Most of the tumours were covered by normal skin, one showed ulceration. Ultrasonography of the tumours allowed accurate anatomical localization in relation to the adjoining tis...
Multisystemic, eosinophilic, epitheliotropic disease with intestinal lymphosarcoma in a horse. Multisystemic, eosinophilic, epitheliotropic disease and intestinal lymphosarcoma were diagnosed in a Paso Fino mare that presented with anorexia and weight loss. The stomach, ileum, cecum, colon, pancreas, and lungs were infiltrated by large numbers of eosinophils forming prominent eosinophilic granulomas, as well as lymphocytes and plasma cells. Two jejunal masses composed of solid sheets of neoplastic lymphocytes were present. In contrast to the regions of inflammation, the infiltrates in these masses did not contain plasma cells, eosinophils, and eosinophilic granulomas. Immunohistochemica...
The distribution of cartilage oligomeric matrix protein (COMP) in tendon and its variation with tendon site, age and load. A protein prominent in guanidine hydrochloride extracts of adult bovine and equine digital flexor tendons was confirmed to be Cartilage Oligomeric Matrix Protein (COMP) by non-reducing and reducing SDS-PAGE, reaction with rabbit anti-COMP polyclonal antiserum on Western blots, trypsin digestion followed by HPLC on a C2/C18 column, and identification of COMP mRNA from tendon on Northern blots. Immunohistochemistry and Western blots of extracts showed COMP to be present in all regions of digital flexor tendons. Equine tendon COMP was purified by ion exchange chromatography and gel filtration and...
Comparative immunohistochemical study of stellate cells in normal canine and equine adenohypophyses and in pituitary tumours. The presence and distribution of S100 protein (alpha and beta subunits), cytokeratin polypeptides, glial fibrillary acidic protein, neurofilaments, vimentin, neuron specific enolase, synaptophysin, HLA class II DR antigen, and pituitary hormones (prolactin, adrenocorticotropic hormone and human chorionic gonadotrophin) in stellate cells were studied immunohistochemically in four normal canine pituitary glands, five canine pituitary adenomas, two canine pituitary carcinomas and two equine pituitary adenomas (with surrounding normal glandular tissue). Stellate cells of the pars distalis and pars...
Neurotrophin receptor-like proteins in Peyer’s patches. The neurotrophins are a family of growth factors that act on responsive cells through specific high-affinity signal-transducing receptors called Trk (A, B, and C) proteins. The neurotrophin receptor proteins are widely distributed in both nervous and nonnervous tissues, including the lymphoid organs. The expression of these receptor proteins by a cell population is an indication of responsiveness to the respective binding neurotrophin. The present study investigated the presence and cellular localization of high-affinity neurotrophin receptor proteins in equine and bovine Peyer's patches. Peye...
Pathogenesis and clinical signs of equine herpesvirus-1 in experimentally infected ponies in vivo. Equine herpesvirus-1 (EHV-1) causes respiratory disease, neonatal death, abortion and neurologic disease. The main purpose of this study was to identify viral antigen in respiratory tract samples by immunoperoxidase staining. Six pony foals were selected on the basis of demonstrating seronegativity to EHV-1 by virus neutralization and housed in isolation. They were infected experimentally by administering EHV-1 nebulized ultrasonically through a face mask. Successful infection was clinically apparent as each of the foals had febrile responses, nasal discharge, and enlarged submandibular lymph ...
Androgen receptors and skeletal muscle composition in trotters treated with nandrolone laureate. To study the effects of nandrolone laureate (19-nortestosterone) on muscle hypertrophy and concentration of androgen receptors (AR), biopsy specimens were taken from the middle gluteal muscle of 6 Finnhorse trotters (geldings and mares) undergoing training before, immediately after, and 13 weeks after a 14-week treatment with nandrolone. Another 6 similarly trained horses served as controls. An additional 10 mares and 10 geldings were used to study annual variation in muscle concentration of AR. AR was immunohistochemically localized in the nuclei. AR concentration remained constant during the...
A sensitive electrophoretic method for the quantification of myosin heavy chain isoforms in horse skeletal muscle: histochemical and immunocytochemical verifications. In adult horses, three myosin heavy chain (MyHC) isoforms can be identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunohistochemistry using specific anti-MyHC monoclonal antibodies. This report studies the suitability of a consistent SDS-PAGE technique for quantifying MyHC profiles in homogenized cryostate sections of equine gluteus medius muscle biopsies (n = 18). The method used (previously described by R. J. Talmadge and R. R. Roy; J. Appl. Physiol. 1993, 75, 2337-2340) resolved MyHCs in three bands: I, IIB or IIX, and IIA from the fastest to the slowe...
Expression of transforming growth factor-beta 1 in normal and dyschondroplastic articular growth cartilage of the young horse. This study describes the distribution pattern of transforming growth factor-beta 1 (TGF-beta 1) mRNA and protein in normal pre- and post natal growth cartilage and alterations present in lesions of dyschondroplasia (osteochondrosis). TGF-beta 1 expression and immunoreactivity have been investigated by in situ hybridisation and immunolocalisation in the articular/epiphyseal growth cartilage of the lateral trochlear ridge of the distal femur. Cartilage was obtained from 19 normal Thoroughbred horses (5 prenatal and 14 post natal horses) and 15 post natal horses with dyschondroplasia (DCP). TGF-b...
Immunohistochemical demonstration of African horse sickness viral antigen in formalin-fixed equine tissues. The distribution of viral antigen was studied in various tissues of three ponies, aged 3-4 years, infected experimentally with a virulent strain of African horse sickness virus (AHSV) serotype 4. Tissues were collected from the animals in the terminal stage of the peracute form of the disease and from one noninfected horse, included as a control. A polyclonal antibody with specificity for AHSV, plus the nonstructural protein NS2, was used in a sensitive avidin-biotin-peroxidase-complex (ABC) method performed on formalin-fixed, paraffin-embedded tissue sections. AHSV antigen was located primari...
Immunocytochemical localization of prolactin and growth hormone in the equine pituitary. The ultrastructural and immunoreactive staining characteristics of cells containing prolactin (lactotropes) and growth hormone (GH; somatotropes) in the anterior pituitaries of gonadally intact pony mares were studied at the electron microscopic level. Lactotropes included two morphological subsets: Type I cells were larger and contained large, dense, polymorphic granules that were scattered throughout the cytoplasm; Type II cells were smaller and contained small, dense, polymorphic granules that were predominantly found in peripheral areas of the cytoplasm. Lactotropes constituted 5 to 16% of...
Immunolocalization of 3 beta-hydroxysteroid dehydrogenase, cytochrome P450 17 alpha-hydroxylase/17,20-lyase and cytochrome P450 aromatase in the equine corpus luteum of dioestrus and early pregnancy. The onset of equine chorionic gonadotrophin (eCG) secretion in pregnant mares is associated with an increase in luteal androgen and oestrogen production. The luteal cell type(s) responsible for the increased production of androgens and oestrogens has not been identified in the equine corpus luteum. In this study, we examined the pattern of expression of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), cytochrome P450 17 alpha-hydroxylase/17,20-lyase (P450(17 alpha)) and cytochrome P450 aromatase (P450arom) by immunohistochemistry in equine luteal tissue collected during dioestrus (days 7-10; ...
The soft-tissue components of the vomeronasal organ in pigs, cows and horses. The soft-tissue components of the vomeronasal organ of the pig, the cow and the horse were studied with the aid of dissection, microdissection, and light microscopy and immunohistochemistry of series of transverse sections. In horses, the rostral end of the incisive duct was blind: thus, unlike in pigs and cows, there was no communication between the vomeronasal organ and the oral cavity. In all three species, the central part of the vomeronasal duct bore the 'typical' respiratory/ receptor epithelium lining on its lateral and medical walls. The rostral part of the duct was characterized by st...
Use of transabdominal ultrasound-guided amniocentesis for detection of equid herpesvirus 1-induced fetal infection in utero. To evaluate transabdominal ultrasound-guided amniocentesis for detection of equid herpes-virus 1 (EHV-1)-induced fetal infection in utero. Methods: 4 Welsh Mountain mares. Methods: Pregnant mares were inoculated intranasally with EHV-1 during the ninth month of gestation. Amniocentesis was initiated on postinoculation day (PID) 12, and was performed at 2- to 3-day intervals in standing mares under deep sedation. Amniotic fluid samples were tested by virus isolation (VI), polymerase chain reaction (PCR), and immunoperoxidase cytologic examination (IC) for detection of EHV-1. Results: Exposure t...
