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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.
Extra-adrenal paraganglioma of the equine orbit: six cases. At The Ohio State University from 1994-2006 six of seven horses evaluated for primary orbital disease were diagnosed with extra-adrenal paraganglioma (EAPG). The horses ranged in age from 14 to 24 years, with a mean of 16.8 years. Duration of clinical signs was 1.5 years to 5 years, with a mean of 2.8 years. Clinical signs varied, but all six had non-painful exophthalmus of the right eye. Five horses had complete ocular exams reported; three of five had decreased to absent vision, two of five had pale optic nerves, and in three of five, difficulty of retropulsion of the globe was noted. Diagno...
Rational design of synthetic peptides to generate antibodies that recognize in situ CD11c(+) putative dendritic cells in horse lymph nodes. A three-dimensional model of the alphaX I-domain of the horse integrin CD11c from dendritic cells provided information for selecting two segments of the primary structure for peptide synthesis. Peptide 1 contains 20 amino acids and peptide 2 has 17 amino acid residues. The first spans from position Thr229 to Arg248 of an alpha-helix segment of the structure, whereas peptide 2 goes from Asp158 to Phe174 and corresponds to an exposed segment of the loop considered to be the metal ion-dependent adhesion site. Murine polyclonal antisera against both peptides were generated and assayed in periphera...
Expression of toll-like receptor 9 in horse lungs. Toll-like receptor 9 (TLR9) has been found to be the main receptor to respond to bacterial DNA in a wide variety of species. Recent work has shown that TLR9 is expressed in a diverse set of cells within the lung. However, much of this data has been centered on human and mouse cell culture lines or primary cultures and very little is known of TLR9 expression in intact lung, especially that of the horse. Here we show that TLR9 is expressed in the lungs of horses in a wide variety of cells. In particular, we note expression in pulmonary intravascular macrophages (PIMs), alveolar macrophages, bron...
Use of contrast-enhanced computed tomography to assess angiogenesis in deep digital flexor tendonopathy in a horse. We compared contrast-enhanced computed tomography (CT) and high field magnetic resonance (MR) imaging in a horse with deep digital flexor tendonopathy. Lesions in the distal extremity were documented grossly and histopathologically. In contrast-enhanced CT, the deep digital flexor tendon lesions were markedly contrast enhancing with evidence of angiogenesis in the core and dorsal border lesions. The lesion morphology was clearly delineated on MR imaging, but without contrast media angiogenesis cannot be identified. Gross examination, histopathologic examination, and CD31 immunohistochemistry c...
A biphasic malignant mesothelioma of the peritoneum and pleura in a horse. This report describes the macroscopic, histologic, immunohistologic and ultrastructural characteristics ofa biphasic malignant mesothelioma in the peritoneal and pleural cavity of a 13-year-old Icelandic pony mare, which exhibited recurrent ascites clinically. Immunohistology was performed employing multiple monoclonal antibodies against cytokeratins (CK) and vimentin. The ultrastructural examination included the quantitative evaluation of the length to diameter ratio of the microvilli. Post mortem examination revealed a severe ascites and hydrothorax. The serosal surfaces of the peritoneum an...
Histomorphological and immunohistochemical study of angiogenesis and angiogenic factors in the ovary of the mare. Cyclical ovaries of 18 mares were examined histologically and immunohistochemically for vascular endothelial growth factor A and B (VEGF A; VEGF B), angiopoietin1 and 2 (Ang1; Ang2), vascular endothelial growth factor receptor 1 and 2 (VEGF-R1; VEGF-R2), angiopoietin receptor (Tie2) and von Willebrand factor. The most intensive coexpression of the examined factors and receptors was detected in the periovulatory period, when a distinctive ovarian angiogenesis takes place, being essential for tertiary follicle maturation and for the endocrine function of the Corpus luteum. Based on the immunohis...
Localized, plexiform, diffuse, and other variants of neurofibroma in 12 dogs, 2 horses, and a chicken. In humans, neurofibroma and schwannoma are distinct entities within the group of benign peripheral nerve sheath tumors. In the veterinary literature, these tumors are often classified together simply as benign peripheral nerve sheath tumors, and diagnostic criteria for their subclassification are not well established. We describe peripheral nerve sheath tumors with microscopic, immunohistologic, and ultrastructural features similar to those in subtypes of human neurofibroma in 12 dogs, 2 horses, and 1 chicken. Dogs were of different breeds and were aged 2 months to 15 years. The canine tumors ...
Expression of prostate glycoconjugates in the stallion and castrated horse. This work was undertaken to determine the glycoconjugates secreted by the epithelium of the prostate in the intact stallion and castrated horse using lectin histochemical procedures in conjunction with enzymatic digestion and deglycosylation treatments. Additionally, anti-5 and 13-16-cytokeratin antibodies were used to localize epithelial basal cells. In the stallion, lectin histochemistry showed the following sugar residues in the Golgi zone of the glandular cells: α-Glu/Man, α-Fuc and β-Gal included in both O- and N-linked oligosaccharides as well as β-GalNAc, GlcNAc and α-Gal, which be...
An atypical equine gastrointestinal stromal tumor. A 17-year-old, gelded Quarter Horse cross was found to have a large, intra-abdominal mass. Clinical signs included infrequent mild colic, weight loss, and chronic anemia. Surgery revealed a very large, discrete, hemorrhagic, multilobular mass with vascular attachments to the transverse colon, mesocolon, jejunal mesentery, and omentum; the site of origin was the transverse colon. Histologic examination demonstrated dense sheets, fascicles, palisades, and interconnecting streams of neoplastic spindle cells with lesser numbers of admixed multinucleated giant cells. Based on morphology alone, this...
Equine thrombospondin II and secreted protein acidic and cysteine-rich in a model of normal and pathological wound repair. Wound healing in horses is complicated, particularly when wounds are on the limb. The objectives of this study were to clone equine thrombospondin II (THBS2) and secreted protein acidic and cysteine-rich (SPARC) cDNAs and to compare the spatiotemporal expression of mRNAs and proteins during repair of body and limb wounds. These molecules were targeted in view of their potential biological contribution to angiogenesis, which is exacerbated during the repair of limb wounds in horses. Cloning was achieved by screening size-selected cDNA libraries previously derived from 7-day-old wounds. Expressi...
Immunohistochemical analysis of laryngeal muscles in normal horses and horses with subclinical recurrent laryngeal neuropathy. We used immunohistochemistry to examine myosin heavy-chain (MyHC)-based fiber-type profiles of the right and left cricoarytenoideus dorsalis (CAD) and arytenoideus transversus (TrA) muscles of six horses without laryngoscopic evidence of recurrent laryngeal neuropathy (RLN). Results showed that CAD and TrA muscles have the same slow, 2a, and 2x fibers as equine limb muscles, but not the faster contracting fibers expressing extraocular and 2B MyHCs found in laryngeal muscles of small mammals. Muscles from three horses showed fiber-type grouping bilaterally in the TrA muscles, but only in the le...
Equine lumican (LUM) cDNA sequence and spatio-temporal expression in an experimental model of normal and pathological wound healing. The development of exuberant granulation tissue, a situation that in some ways resembles the human keloid, compromises both the aesthetic and functional outcomes of wound repair in horses. To help elucidate the underlying molecular mechanisms the spatio-temporal expression of lumican (LUM) mRNA and protein for their potential contributions to tissue remodelling of body and limb wounds, was examined in an established experimental model. Expression was studied in intact skin and in samples of 1-, 2-, 3-, 4- and 6-week-old wounds of the body and forelimb. Temporal gene expression was determined b...
Neurokinin receptors in recurrent airway obstruction: a comparative study of affected and unaffected horses. The purpose of the study was to compare in vitro airway responses to neurokinin A & B (NKA and NKB) and expression of NK-2 receptors in airways of horses affected and unaffected with recurrent airway obstruction (RAO). Neurokinin-A, an inflammatory mediator belonging to the tachykinin family of neuropeptides, causes bronchoconstriction by binding to NK-2 receptors. Neurokinin-B is a lesser-known neuropeptide that acts on NK-3 receptors. Horses were placed into RAO-affected and RAO-unaffected groups based on their history, clinical scoring, and pulmonary function testing. Lung tissue from e...
P-glycoprotein in intestines, liver, kidney and lymphocytes in horse. P-glycoprotein (P-gp) is an important drug transporter, which is expressed in a variety of cells, such as the intestinal enterocytes, the hepatocytes, the renal tubular cells and the intestinal and peripheral blood lymphocytes. We have studied the localization and the gene and protein expression of P-gp in these cells in horse. In addition we have compared the protein sequence of P-gp in horse with the protein sequences of P-gp in several other species. Real time RT-PCR and Western blot showed gene and protein expression of horse P-gp in all parts of the intestines, but there was no strict cor...
Clara cell secretory protein is reduced in equine recurrent airway obstruction. Horses are prone to recurrent airway obstruction (RAO), an inflammatory lung disease induced by repeated exposure to environmental mold, dust, and bacterial components. Active disease manifests with mucus hyperproduction, neutrophilic inflammation, bronchoconstriction, and coughing. Chronically affected animals have lung remodeling characterized by smooth muscle hyperplasia, collagen deposition, lymphoid hyperplasia, and impaired aerobic performance. Clara cell secretory protein (CCSP) counters inflammation in the lung, hence we hypothesized that CCSP depletion is a key feature of RAO in horse...
Activated platelet-derived growth factor beta receptor expression, PI3K-AKT pathway molecular analysis, and transforming signals in equine sarcoids. The equine sarcoid is the most common dermatologic neoplasm reported in horses. Bovine papillomavirus (BPV) types 1 and 2 are associated with sarcoids, in which the expression of the major transforming oncoprotein (E5) is often recorded. The transformation activity of the virus is due to the binding of the E5 to the platelet-derived growth factor beta receptor (PDGFbeta-r). In the present study, we show by Western blot in 4 sarcoid samples and 3 normal equine skin samples that the PDGFbeta-r is more phosphorylated in sarcoid tissue than in normal skin (P < .001). Furthermore, the physical i...
Identification of Bartonella henselae in an aborted equine fetus. This report describes the characterization of a Bartonella henselae abortion in an equine fetus by gross, histologic, immunohistochemical, ultrastructural, and molecular methods. Bartonella henselae can cause cat scratch disease, bacillary angiomatosis, bacillary peliosis, and endocarditis in humans and other animals. The bacterium has been isolated from several mammalian species but only recently from equids; however, it has not been linked to abortion in equids. An aborted equine fetus exhibited necrosis and vasculitis in multiple tissues, with intralesional Gram-negative short-to-spirillar ...
Bilateral intraocular glandular choristomas in a Thoroughbred foal. Intraocular choristomas are rare anomalies in domestic animals and are often associated with multiple ocular malformations. A Thoroughbred foal presented for ocular abnormalities and was diagnosed with microphthalmia, corneal dermoids, severe anterior segment dysgenesis (including glandular choristomas), aphakia, retinal dysplasia, and optic nerve hypoplasia. Morphological, histochemical, and immunohistochemical comparisons were made between ocular choristomatous tissues from this foal and lacrimal gland, third eyelid gland, nasopharynx, trachea, and lacrimal sac/nasolacrimal duct from normal ...
Neuropathology of naturally occurring Trypanosoma evansi infection of horses. The clinical signs and pathology of the central nervous system in 9 horses with naturally occurring neurologic disease due to Trypanosoma evansi are described. The clinical course was 2 to 20 days; clinical signs included marked ataxia, blindness, head tilt and circling, hyperexcitability, obtundity, proprioceptive deficits, head pressing, and paddling movements. Grossly, asymmetric leukoencephalomalacia with yellowish discoloration of white matter and flattening of the gyri were observed in the brain of 7 of 9 horses. Histologically, all 9 horses had necrotizing encephalitis that was most sev...
Histologic evaluation of the diversity of epidermal laminae in hooves of horses without clinical signs of laminitis. To evaluate the histologic diversity of epidermal laminae in hooves from horses without clinical signs of laminitis. Methods: Formalin-fixed samples of stratum internum obtained from the mid region of the dorsal aspect of the hoof wall from the forelimbs of 35Thoroughbred cadavers (including foals [n = 9], yearlings [5], 2 year olds [6], racing horses [5], and mares [10]). Methods: Paraffin-embedded laminar tissues were stained with H&E for the evaluation of architectural variety of primary epidermal laminae (PEL) and secondary epidermal laminae (SEL). For detection of cytokeratin (CK) exp...
Biological and anatomical evidence for kisspeptin regulation of the hypothalamic-pituitary-gonadal axis of estrous horse mares. The purpose of the present study was to evaluate the effects of kisspeptin (KiSS) on LH and FSH secretion in the seasonally estrous mare and to examine the distribution and connectivity of GnRH and KiSS neurons in the equine preoptic area (POA) and hypothalamus. The diestrous mare has a threshold serum gonadotropin response to iv rodent KiSS decapeptide (rKP-10) administration between 1.0 and 500 microg. Administration of 500 microg and 1.0 mg rKP-10 elicited peak, mean, and area under the curve LH and FSH responses indistinguishable to that of 25 microg GnRH iv, although a single iv injection...
Identification of protein tyrosine phosphatases and dual-specificity phosphatases in mammalian spermatozoa and their role in sperm motility and protein tyrosine phosphorylation. Protein tyrosine kinases have important roles in spermatozoa; however, little is known about the presence and regulation in these cells of their counterparts in signaling, namely, protein tyrosine phosphatases (PTPs) and dual-specificity phosphatases (DSPs). The objectives of the present study were to identify PTPs and DSPs in boar, stallion, and dog spermatozoa; to characterize their subcellular distribution; and to investigate the roles of tyrosine phosphatases in maintenance of protein tyrosine phosphorylation level and in sperm motility. Using Western blotting with specific antibodies in b...
Calprotectin in myeloid and epithelial cells of laminae from horses with black walnut extract-induced laminitis. Laminar inflammation is one of the earliest events in equine laminitis. Calprotectin (CP), a Damage-Associated Molecular Pattern protein, is overexpressed in inflammatory conditions of human skin. Objective: CP is overexpressed in the laminar epidermis of horses with black walnut extract (BWE)-induced laminitis. Methods: Twenty adult horses. Methods: Experimental study. Horses were allocated to one of 4 groups. BWE was administered to horses in 3 groups, which were sampled 1.5, 3, and 12 hours (LAM) later. CP was visualized by immunohistochemistry. Laminar leukocyte counts and intensity of lam...
Pleomorphic corneal sarcoma resembling malignant peripheral nerve sheath tumor in a horse. A 14-year-old Haflinger gelding presented with a protruding mass involving the cornea of the right eye. The mass was resected and submitted for histopathologic and immunohistochemical examination. The preliminary diagnosis was corneal sarcoma, most likely fibrosarcoma. The immunohistochemical results confirmed the mesenchymal origin of the neoplastic cells, which were most consistent with a malignant peripheral nerve sheath tumor. Corneal mesenchymal neoplasms are extremely uncommon tumors in human beings and domestic animals. The cause for this tumor was not determined; infection with bovine ...
OB-cadherin cloning and expression in a model of wound repair in horses. Horses suffer from a debilitating impediment in repairing wounds located on the lower limb that leads to the development of a fibroproliferative disorder (exuberant granulation tissue). This condition is a source of wastage since it often forces retirement from competition. Treatments that resolve or prevent this condition are still lacking, maybe due to deficient knowledge of the underlying molecular mechanisms. Fibroblast-to-myofibroblast conversion is an essential step allowing contraction during wound repair and is accompanied by an increase in OB-cadherin expression. Objective: To clone e...
Corneal stromal invasive squamous cell carcinoma: a retrospective morphological description in 10 horses. To describe the pathomorphological features of corneal stromal invasive squamous cell carcinoma (CSI-SCC) in horses. Methods: A total of 87 equine SCC in the Comparative Ocular Pathology Laboratory of Wisconsin database were retrieved. The signalment and anatomical distribution were summarized. Ten CSI-SCC out of 87 SCCs were further investigated focusing on pathomorphological description. All 10 cases were stained with H&E, periodic acid-Schiff stain and Verhoeff's elastic stain. Results: Four Appaloosas, two Quarter horses, two American Paint, one Pinto and one Thoroughbred horse were af...
Expression of cyclooxygenase-2 by equine ocular and adnexal squamous cell carcinomas. To investigate whether cyclooxygenase-2 (COX-2) is expressed by equine ocular and adnexal squamous cell carcinomas (SCC). Methods: Forty-three samples of histologically confirmed cases of ocular SCC or carcinoma in situ (CIS) from 34 horses presented to the Animal Health Trust between 1992 and 2004 were subjected to a standard, two-layered, indirect immunohistochemical method using a rabbit polyclonal antihuman COX-2 antibody. Ten formalin-fixed, paraffin-embedded tissue samples taken from recognized predilection sites for SCC, from the grossly normal eyes of 10 horses euthanized for reasons u...
Poorly differentiated leiomyosarcoma of the urogenital tract in a horse. A 2-year-old Thoroughbred filly was evaluated because of hemorrhage from the vulva and suspected hematuria of 5 days' duration. Results: A primary coagulopathy was ruled out on the basis of results of hematologic testing. Vaginoscopy and cystoscopy revealed a large bleeding mass in the bladder that extended into the vagina, causing marked obliteration of normal urogenital structures and difficulty in urination. Histologic examination of endoscopic and surgical biopsy specimens revealed a poorly differentiated neoplasia likely of mesenchymal origin. Chronic suppurative cystitis caused by Strept...
Laminin receptor 1 is differentially expressed in thoracic and limb wounds in the horse. Healing of wounds located on the distal limbs of horses is often complicated by retarded epithelialization and the development of exuberant granulation tissue (proud flesh). Treatments that definitively resolve this pathological process are still unavailable. Molecular studies of the repair mechanism might contribute to the development of new therapeutic strategies. The study presented herein aimed to clone the full length cDNA and to study the spatio-temporal expression profile of mRNA and protein for LAMR1, previously attributed a role in wound epithelialization, during the repair of body an...
