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.
Pollitt CC.In the equine hoof, the basement membrane connects the heavily keratinised hoof wall to the dense connective tissue of the distal phalanx, a region able to withstand considerable mechanical stress. This study investigated the properties of this important anatomical and physiological structure. In contrast to haematoxylin and eosin, the connective tissue stains, periodic acid Schiff, periodic acid silver methenamine and Azan showed good resolution of lamellar basement membrane. The lamellar basement membrane cross-reacted with mouse monoclonal antibodies raised against human laminin, thereby pr...
Amselgruber WM, Sinowatz F, Erhard M.The present study deals with the immunohistochemical localization of S-100 protein in the testes of seven mammalian species including rat, cat, dog, pig, sheep, cattle and horse. Significant differences are demonstrated in the cellular distribution and intensity of immunoreaction for the protein. In bull, ram, boar and cat testes S-100 protein was localized in the cytoplasm and nuclei of Sertoli cells. A particularly intense staining was seen in the modified Sertoli cells of the terminal tubular segment. With the exception of the cat and horse S-100 protein immunoreactivity was additionally fo...
Tanimoto T, Yamasaki S, Ohtsuki Y.A well-demarcated solitary splenic mass (20 x 20 x 15 cm in size) containing hemorrhagic and necrotic foci was observed in a 4-year-old Thoroughbred stallion. Histologically, the mass consisted of lymphoma cells of the diffuse large non-cleaved type, with a high mitotic index and scattered macrophages that formed a starry sky pattern. The lymphoma cells revealed diffuse positivity for acid phosphatase and alpha naphthyl butyrate esterase, and were also positive for intracytoplasmic IgM on occasion, and mostly for proliferating cell nuclear antigen. Ultrastructural examination revealed moderate...
Sonea IM, Bowker RM, Robinson NE, Broadstone RV.Distribution of pulmonary nerves immunoreactive for either substance P or calcitonin gene-related peptide was determined, using immunohistochemical methods on healthy lungs from adult equids. The overall patterns of distribution of substance P and calcitonin gene-related peptide-like immunoreactivity were similar. Distribution of immunoreactive nerves was not uniform throughout the lungs; nerve fibers immunoreactive for these peptides were more frequently observed near the hilus of the lung than in the caudal lobes or in the periphery of the lung. Nerve fibers immunoreactive for substance P or...
Blanchard-Channell M, Moore PF, Stott JL.Two monoclonal antibodies (mAb), UC F6G-3 and UC F13C-5, were characterized as being specific for the apparent equine homologues of CD3 and CD5, respectively. Both antibodies exhibited characteristics of pan-T-lymphocyte markers based upon immunohistology and two-colour flow cytometry. UC F6G-3 precipitated a complex of proteins (up to seven) with molecular weights ranging from 18,000 to 42,000, similar to the human and murine CD3 complex. Upon further dissociation of the precipitated complex, two proteins were identified with molecular weights of 22,000 and 27,000. Immobilized UC F6G-3 was ef...
Oikawa M, Kamada M, Yoshikawa Y, Yoshikawa T.Seven horses that died of pneumonia associated with transport yielded Streptococcus equi subsp. zooepidemicus (S.z.) from their pulmonary lesions. These lesions were divisible roughly into two types, serous haemorrhagic pneumonia and multiple foci of coagulative necrosis, which were considered to reflect a temporal difference in the process of lesion formation. Immunohistologically, S.z. antigen was detected in both types of lesion. Acute necrotic lacunar tonsillitis was considered to play an important role in the onset of the pneumonia.
Zhang CH, Donaldson WL, Antczak DF.A surface antigen of equine B lymphocytes was identified using the Equine Leucocyte Antigen Workshop antibody WS 65. This marker was expressed on almost all equine B cells, but not on T cells, granulocytes or thymocytes. WS 65 strongly stained cells in the follicular areas of lymph nodes and cells in the splenic nodules when tested on frozen tissue sections by immunohistochemistry. Equine leukemic T cells were not labeled by WS 65, and neither were the cells from a horse with B cell leukemia, although these latter cells carried surface immunoglobulin. Immunoprecipitation of lymphocyte membrane...
Rosol TJ, Nagode LA, Robertson JT, Leeth BD, Steinmeyer CL, Allen CM.Humoral hypercalcemia of malignancy was evident in a horse that had a locally invasive ameloblastoma of the left hemimandible. Surgical removal of the neoplasm resulted in prompt return of serum calcium and parathyroid hormone concentrations to within reference limits. The tumor contained parathyroid hormone-related protein, as demonstrated by immunohistochemistry and western blot analysis. It is likely that production of this protein by the neoplasm was important in the pathogenesis of the hypercalcemia. The case represented a sporadic form of humoral hypercalcemia of malignancy attributable ...
Pearson GT.The architecture and neurochemistry of the enteric nervous system was studied by use of whole-mount preparations obtained by microdissection of the horse jejunum. A myenteric plexus and two plexuses within the submucosa were identified. The external submucosal plexus lying in the outermost region of the submucosa had both neural and vascular connections with the inner submucosal plexus situated closer to the mucosa. Counts of neurones stained for NADH-diaphorase demonstrated the wide variation in size, shape and neurone content of individual ganglia in both the external and internal submucosal...
Kim DY, Hodgin EC, Lopez MK, Nasarre C.A large primary retroperitoneal sublumbar neoplasm in a horse, with disseminated neoplastic foci in the brain, lung, kidney and spleen is described. The diagnosis was based on light microscopical studies and positive immunostaining for neuron-specific enolase. Because of the location of the primary tumour mass, the aortico-sympathetic ganglion (organ of Zukerkandl) is proposed as the origin.
Prieto D, Simonsen U, Martín J, Hernández M, Rivera L, Lema L, García P, García-Sacristán A.The distribution of acetylcholinesterase (AChE)-positive nerve fibers and cells, as well as the effects of acetylcholine (ACh) on ureteral smooth muscle and small resistance arteries were investigated in the equine ureter by means of histochemical, classic organ baths and myograph techniques. AChE-positive nerve fibers were widely distributed throughout the ureteral wall forming muscular, subepithelial and perivascular nerve plexuses, whose density was highest at the intravesical ureter. AChE-positive nerve cells were also identified grouped as adventitial or intramural ganglia. ACh increased ...
Wohlsein P, Harps O, Trautwein G.The clinical history, clinical, pathological and immunohistological findings of a four-month-old foal with generalised pemphigus foliaceus are presented. The typical lesions of this autoimmune skin disease are described and discussed.
Singh B, Minhas KJ, Atwal OS.The pulmonary intravascular macrophages (PIMs) of the ponies possess all the characteristics of a fully differentiated resident macrophage, which forms adhesive plaques with the capillary-endothelial cells. In addition, it has a unique surface coat which does not conform to the concept of a carbohydrate rich membrane bound glycocalyx generally associated with macrophages. We studied the responses of these cells especially in the context of its globular surface coat to multiple doses of MB intravenously: 0.2 ml/kg body copper tracer substance. The ponies were treated with three doses of MB intr...
Cobb MA, Schutt WA, Hermanson JW.The horse provides an interesting model for study of the structure and function of the mammalian diaphragm. Multiple regions of diaphragm from seven adult horses were prepared for histochemistry, immunocytochemistry, myosin heavy chain electrophoresis, and native myosin electrophoresis. Two additional adults were dissected to demonstrate myofiber and central tendon morphology and stained for acetylcholinesterase to demonstrate motor endplates. All regions of the adult diaphragm were histochemically characterized by a preponderance of type I fibers with some type IIa fibers. Type IIb fibers wer...
Kuwamura M, Iwaki M, Yamate J, Kotani T, Sakuma S, Yamashita A.A 9-year-old male horse showed emaciation, weakness and trembling and was euthanatized. Histopathological examinations revealed loss, swelling and chromatolysis of motor neurons throughout the spinal ventral horns, axonal degeneration of the ventral spinal roots. Eosinophilic cytoplasmic inclusions were distributed in degenerated spinal ventral neurons. Ultrastructurally, the inclusions consisted of aggregations of granular dense material and a few vesicles. They reacted positively with polyclonal antibody against ubiquitin. The present case was diagnosed as equine motor neuron disease, which ...
Nixon AJ, Cummings JF.Serial sections of bone and soft tissue from the metacarpophalangeal joints of 2 mature and 2 immature horses were evaluated for substance P immunoreactive sensory nerve fibers. Formalin-fixed specimens were sectioned, either nondemineralized or demineralized with formic acid or EDTA. Rabbit antiserum to substance P (SP) was used in the streptavidin-biotin-peroxidase complex method for immunolocalization of SP antigen, and staining with 3,3'-diaminobenzidine was used for permanent identification of SP fibers. Abundant sensory nerve fibers were identified in the joint capsule, synovial membrane...
Oriol JG, Sharom FJ, Betteridge KJ.The embryonic capsule, which covers the equine blastocyst after it loses its zona pellucida, is composed of mucin-like glycoproteins. In the present study, we investigated both macroscopic and molecular changes in the capsule during development. The weight of the capsule increased from day 11-12 of pregnancy and reached a maximum at about day 18, coinciding with the time during which the conceptus migrates extensively throughout the uterus. The sialic acid content of the capsule declined markedly from about day 16, the time of conceptus 'fixation' in the uterus, which suggests a unique develop...
Hamir AN, Moser G, Galligan DT, Davis SW, Granstrom DE, Dubey JP.A 5-year (1985-1989) retrospective immunohistochemical study was conducted using an avidin-biotin complex (ABC) immunoperoxidase method to demonstrate Sarcocystis neurona in histologically suspect cases of equine protozoal myeloencephalitis (EPM). Primary antibodies against S. neurona and S. cruzi were utilized for the ABC technique. The findings were compared with those from cases in which the organisms were detected by examination of hematoxylin and eosin (HE)-stained neuronal sections. HE-stained sections detected the presence of the organisms in 20% of the suspect cases; whereas the ABC te...
Fukunaga K, Ninomiya M, Oohara Y, Kusunose K, Okamura Y, Nagasaki H, Ishino S, Kadota K.Lymphoplasmacytic lymphoma found in a 6-year-old Anglo-Arabian stallion was investigated histologically, immunohistochemically and ultrastructurally. The animal showed a large mediastinal mass and generalized lymph node involvement. The neoplastic cells were in various differentiation stages of small lymphocyte, centrocyte, centroblast, immunoblast and plasma cell. Some neoplastic cells showed positive cytoplasmic reactivity for mu and lambda chains. There were well developed rough endoplasmic reticulum (RER) and Golgi complexes in plasmacytoid cells, and slightly developed RER or a few long s...
Prieto D, Hernandez M, Rivera L, Ordaz E, Garcia-Sacristan A.The aim of the present study was to determine the distribution and density of catecholaminergic nerve fibres and cells in the equine ureter by using immunohistochemical techniques to localise the enzyme tyrosine-hydroxylase (TH). TH-immunoreactive (TH-IR) nerve fibres entered the wall of the ureter as adventitial nerve trunks accompanying the blood vessels. These trunks repeatedly branched as they coursed through the muscular layer towards the epithelium, forming muscular, perivascular and subepithelial nerve plexuses. TH-IR nerve fibres were especially numerous in the pelvic and intravesical ...
Sonea IM, Bowker RM, Broadstone RV, Robinson NE.The tone of respiratory smooth muscle is largely determined by the input from autonomic nerves. The distribution of adrenergic and selected nonadrenergic, non-cholinergic (NANC) nerves in the normal equine trachealis muscle was investigated using immunohistochemistry. The smooth muscle of the trachealis was found to contain numerous nerves immunoreactive for an enzymatic marker of adrenergic nerves, as well as many nerves immunoreactive for a putative NANC neurotransmitter, peptide histidine isoleucine, a potent bronchodilator. The tissue surrounding the respiratory smooth muscle contained num...
Sellon DC, Cullen JM, Whetter LE, Gebhard DH, Coggins L, Fuller FJ.An IgG1 mouse monoclonal antibody, designated 1.646, is described which recognizes a cytoplasmic antigen of equine mononuclear phagocytes. Indirect fluorescent antibody staining of peripheral blood leukocytes reveals a granular cytoplasmic staining, predominantly in adherent blood mononuclear cells. Indirect fluorescent antibody staining is positive for alveolar and peritoneal macrophages. In some horses, a few neutrophils are also stained. In equine tissue samples stained by immunohistochemistry, the distribution of positive cells is consistent with the distribution of tissue macrophages. The...
Sasaki K, Igarashi S, Amasaki T, Amasaki H, Nishita T, Kano Y, Asari M.Immunohistochemical localizations of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in equine and bovine digestive tracts were studied. In the horse, epithelial cells in both the oesophagus and non-glandular part of the stomach lacked all three isozymes. In contrast, surface epithelial and parietal cells in the glandular region of the stomach showed reactivity for CA-II. In the small intestine, absorptive columnar cells covering the villi in the duodenum were positive for CA-II. The epithelium of the jejunum and ileum lacked all three isozymes. In the large intestine, CA-II was detected ...
Sonea IM, Bowker RM, Robinson NE, Broadstone RV.Immunohistochemical methods were used to determine the distribution of pulmonary nerves containing either an enzymatic marker of adrenergic nerves, dopamine beta hydroxylase, or the putative neurotransmitter neuropeptide Y in 7 equids with healthy lungs. Nerves immunoreactive for these substances were found on airway smooth muscle in nearly all the samples of healthy equine lung examined. These nerves were generally more numerous in the larger airways but could be detected even in noncartilaginous bronchioles. Pulmonary and bronchial vessels also contained numerous immunoreactive nerves. On th...
Rimstad E, Evensen O.Paraffin-embedded organ samples from 28 aborted fetuses and three foals, partly archival and partly sampled in 1991, were examined by polymerase chain reaction (PCR) and immunohistochemistry for the presence of DNA and antigens, respectively, specific for equine herpesvirus 1 (EHV-1). Virologic examination had been performed on 23 of the aborted fetuses. DNA fragments specific for EHV-1 were identified by PCR, and EHV-1 antigens were identified in situ by immunohistochemistry, with an agreement between the methods of 94% (kappa = 0.85). Compared with virus isolation, PCR agreement was 87% (kap...
Sinha AK, Ray SP, Rose RJ.Skeletal muscle adaptations to training of differing intensities were examined in 10 thoroughbred horses that underwent six weeks of treadmill training followed by six weeks of detraining. The horses were randomly assigned to either a slow group exercised at 40 per cent maximum oxygen uptake (VO2max) or a fast group at 80 per cent VO2max. Resting muscle biopsies were taken before training, after six weeks of training and after six weeks of detraining, from m gluteus medius and m biceps femoris. Muscle was analysed histochemically for fibre type composition (myosin ATPase) and capillary supply ...
Watson ED, Dixon CE.The distribution of T lymphocytes and of cells bearing MHC Class II antigens in the endometrium of the mare was studied using an avidin-biotin-peroxidase staining method. The cells within the endometrium which expressed MHC Class II were macrophages, lymphocytes, monocytes, dendritic cells, epithelial cells and endothelial cells. MHC Class II expression increased significantly (P < 0.05) in the luminal epithelium and tended (P = 0.0573) to increase in the subepithelial layers during oestrus. Numbers of T lymphocytes did not differ between oestrus and dioestrus. MHC Class II expression and T...
Patnaik AK.In a retrospective study, granular cell tumors in six dogs (Nos. 1-6), three cats (Nos. 1-3), one horse (No. 1), and one cockatiel (Nymphicus hollandicus) (No. 1) and a meningioma with a granular cell component in one dog (No. 7) were examined histologically and immunohistochemically. These tumors were identified by histologic examination of surgical biopsy specimens, except in the horse, in which the tumor was an incidental finding at necropsy. These diagnoses were initially made by more than one pathologist. Five of the six granular cell tumors in the dogs were in the oral cavity; one of the...
Smith KC, Whitwell KE, Mumford JA, Gower SM, Hannant D, Tearle JP.Twelve Welsh Mountain pony mares in late gestation were infected intranasally with EHV-1 (AB4 isolate) at dose rates from 10(3) to 10(7.3) TCID50. This resulted in 3 cases of paresis, at Days 9, 10 and 12 after inoculation, and 5 abortions, at Days 6, 9, 18, 19 and 20. Euthanasia was performed between Days 6 and 21, with collection of uterine specimens for histopathology, virus isolation and immunoperoxidase staining from the pregnant horn, non-pregnant horn and body. EHV-1 replication in endometrial vessels was detected as early as Day 6 and was maximal at Days 9-11, when widespread thrombois...
Schultheiss PC, Collins JK, Carman J.An indirect immunoperoxidase (IP) procedure using the avidin-biotin-peroxidase complex detection technique was developed to detect viral equine herpesvirus-1 (EHV-1) antigen in formalin-fixed paraffin-embedded tissues from aborted equine fetuses. The procedure was applied to liver, lung, and other tissues from 20 cases of confirmed or suspected EHV-1-induced abortions. Specific staining was observed in tissue sections from EHV-1-infected fetuses. Positive IP staining was present in tissues of 7 cases that were also positive by fluorescent antibody (FA) and virus isolation (VI) and that had typ...
Lee JY, Kang TY, Lee YD, Shin TK.The binding specificities of various lectins, such as the Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), and the Bandeiraea simplicifolia BS-1 (Isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I) lectins, were studied in the vomeronasal organ of the horse. The microvilli of the vomeronasal sensory epithelium were positive for DBA, SBA, Isolectin B4, WGA, PNA, and UEA-I. The receptor cells showed intense reactivity for DBA and WGA. Lectins were not detected in the supporting cells or basal cells. The Jacobson's glands were positive for WGA a...
Poradowski D, Chrószcz A.The study consisted of the immunohistochemical analysis of fundic and pyloric mucosa in the equine stomach between the 4th and 11th month of gestation. The accessible material was classified into three age groups using the CRL method. The adult reference group was used to define potential differences between foetal and adult populations of gastric APUD cells. The samples were preserved, prepared, and stained according to the standard protocols. The immunohistochemical reaction was assessed using the semi-quantitative IRS method. The results were documented and statistically analysed. The most ...
Galeano R, Germanà A, Abbate F, Calvo D, Naves FJ, Hidaka H, Germanà G, Vega JA.Neurocalcin (NC) is a recently characterized EF-hand calcium-binding protein present in a discrete population of sensory neurons and their peripheral mechanoreceptors, but its presence in peripheral nervous system neurons other than in the rat is still unknown. The present study was designed to investigate the occurrence of NC in the dorsal root ganglia (DRG) of several mammalian species (horse, buffalo, cow, sheep, pig, dog, and rat), including humans. DRG were fixed, embedded in paraffin, and processed for immunohistochemistry using a polyclonal antibody against NC. The size of the immunorea...
Baum B, Hewicker-Trautwein M, Wohlsein P.A 10-year-old Knabstrupper stallion was euthanatized because of severe dyspnea and exercise intolerance. Postmortem examination revealed diffuse severe alveolar emphysema and chronic fibrosing pleuritis of the caudal lung. Parts of both caudal lung lobes were covered with multiple raised firm gray to yellow plaques. Histologically, these areas consisted of circumscribed pleural fibroses and cysts of metaplastic keratinizing squamous epithelium. Immunohistochemistry revealed intense labeling for cytokeratins 5/6 and 10. In addition, caudal lung lobes were severely affected by a chronic partiall...
Szeredi L, Makrai L, Dénes B.The first objective of this study was to develop an immunohistochemical procedure for rapid detection of Rhodococcus equi in impression smears from affected organs of foals on postmortem examination. The second aim was to demonstrate whether R. equi can be detected in smears of tracheal exudates collected from the same foals using an immunohistochemical method. Impression smears and cryostat and paraffin-embedded sections were made from the lungs and mediastinal lymph nodes of three foals (A, B and C) that had died of respiratory disease caused by R. equi, and also from the caudal mesenteric l...
Polikepahad S, Haque M, Francis J, Moore RM, Venugopal CS.The purpose of the study was to determine and compare the expression of endothelin (ET) receptors in the peripheral lungs of healthy horses and those affected with recurrent airway obstruction (RAO) using reverse transcriptase polymerase chain reaction (RT-PCR), real-time PCR, Western blot analysis, and immunohistochemical techniques. Two groups of horses (7 healthy and 7 RAO-affected) were selected from a pool of horses destined for euthanasia. The grouping of horses was based on the history, clinical scoring, and pulmonary function testing. After euthanasia, gross postmortem evaluation of th...
Holzhausen L, Nowak M, Junginger J, Puff C.A 15-year-old gelding presented with a progressive lameness of the left forelimb of 2.5 months duration. Clinically, a dilation of the deep flexor tendon sheath with a firm elastic consistency and a pronounced tenderness was noted. Ultrasonically, a marked swelling of the flexor tendon sheath with an irregular density of the mesotendineum was observed. The white, firm material forming a nodular distension of the flexor tendon sheath with a diameter of approximately 1 cm was excised and sent for histopathological examination. Biopsies of the deep flexor tendon and corresponding tendon sheath we...
Respiratory diseases are a major cause of morbidity and mortality in the horses of all ages including foals. There is limited understanding of the expression of immune molecules such as tetraspanins and surfactant proteins (SP) and the regulation of the immune responses in the lungs of the foals. Therefore, the expression of CD9, SP-A and SP-D in foal lungs was examined. Results: Lungs from one day old (n = 6) and 30 days old (n = 5) foals were examined for the expression of CD9, SP-A, and SP-D with immunohistology and Western blots. Western blot data showed significant increase in the...
Eto D, Yamano S, Hiraga A, Miyata H.There is little information about the muscle fibre recruitment pattern during sloped and flat track running in Thoroughbred horses. Objective: To examine the glycogen depletion pattern of each muscle fibre type during running on a flat and sloped treadmill. Methods: Thirteen Thoroughbred horses (3-9 years old) were used. They were initially subjected to incremental exercise tests on a treadmill at 10 and 0% inclines in each horse to determine running speed at 90 and 60% VO2max. Needle biopsy samples were obtained from the middle gluteal muscle immediately after the running at 90% VO2max for 4 ...
Miragliotta V, Lefebvre-Lavoie J, Lussier JG, Theoret CL.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...
Mykkänen AK, Hyyppä S, Pösö AR, Ronéus N, Essén-Gustavsson B.Monocarboxylate transporter 1 (MCT1) and its ancillary protein CD147 facilitate efflux of lactate from the muscle. Expression of MCT1 and CD147 were studied with immunohistochemistry in type I, IIA, IIAB and IIB fibres of equine gluteal muscle. Staining intensity of MCT1 in the cytoplasm as well as in the membranes of fibre types decreased in the order I=IIA>IIAB>IIB and correlated with the oxidative capacity. Capillaries were pronounced in the MCT1 staining. CD147 antibody stained plasma membranes of all fibre types evenly, whereas the staining in the cytoplasm followed that of MCT1. In...
Mobarak MS, Ryan MF.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...
Dall'Aglio C, Mercati F, Pascucci L, Ceccarelli P.The aim of the present study was to demonstrate the presence and the distribution of leptin and its receptor in the pancreas of horses of both sexes by immunohistochemical techniques. The presence and the distribution of leptin receptor were also investigated in the initial portion of the duodenum, near the duodenal ampulla. The immunohistochemical investigation demonstrates the immunolocalization of both leptin and its receptor in the endocrine cells of pancreatic islets, which led us to hypothesize that leptin may possibly exert an autocrine/paracrine action on the endocrine pancreas. Examin...
Bouchard PR, Fortna CH, Rowland PH, Lewis RM.Granular cell tumor (GCT) is a morphologic designation for tumors of varied histogenesis. Most GCTs in human beings are derived from Schwann cells, and rat meningeal GCTs are believed to originate in the neural crest. Three equine pulmonary GCTs from aged horses were studied immunohistochemically with primary antibodies directed against vimentin, cytokeratins (AE1/AE3), S-100, Leu 7, desmin, and neuron-specific enolase (NSE) using a steptavidin-biotin procedure. All three tumors stained similarly with strong and diffuse staining of neoplastic cells for vimentin and S-100 and negative staining ...
Kasperowicz B, Rotkiewicz T, Otrocka-Domagała I.The purpose of the study was a pathomorphological and immunohistochemical analysis of tumour cells and connective tissue in equine sarcoids. Investigations were performed using histopathological, ultrastructural, immunohistochemical (PCNA, p53, cytokeratin, vimentin) and histochemical (Ag-NORs) methods. The study was conducted on 50 sarcoids originating from 36 horses and classified as occult, verrucous, fibroblastic and a mixed type of sarcoid based on their clinical appearance. Most of the tumours were located on the girth (30%), neck (24%), head (12%), and legs (12%). The average age of the...
Nasu T, Sueishi A, Morimoto M.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...
Roels S, Ducatelle R, van De Vijver B, De Kruif A.An 18-month-old cross-bred Fjord pony colt presenting a fibrosarcoma of the prepuce with severe infiltrative growth, was treated by topical application of 5-fluorouracil. Biopsies were taken before and after a treatment period of 2 months. As a control, preputial tissue from a healthy horse was biopsied. Tissue sections were stained immunohistochemically for Proliferating Cell Nuclear Antigen (PCNA) and Proliferation related factor--Ki67 (MIB1). Using computerized quantitative image analysis of these sections, Mitotic index (PCNA), Growth Fraction (Ki67) and total nuclear area percentage of th...
Bombardi C, Grandis A, Gardini A, Sorteni C, Clavenzani P, Chiocchetti R.The activity of the gastrointestinal tract is regulated through the activation of adrenergic receptors (ARs). Since data concerning the distribution of ARs in the horse intestine is virtually absent, we investigated the distribution of β2-AR in the horse ileum using double-immunofluorescence. The β2-AR-immunoreactivity (IR) was observed in most (95%) neurons located in submucosal plexus (SMP) and in few (8%) neurons of the myenteric plexus (MP). Tyrosine hydroxylase (TH)-IR fibers were observed close to neurons expressing β2-AR-IR. Since β2-AR is virtually expressed in most neurons located...
Budras KD, Schiel C, Mülling CK, Patan B.The preparation of hard tissues such as the equine hoof horn for electron microscopic examination is very difficult. In particular the penetration of fixatives and chemicals used during fixation and embedding is a problem. The objective of this study was to find and implement an alternative method enabling the preparation of high-quality thin sections of hoof horn and other hard tissue, which maintains the hard tissue ultrastructure and can be used for immuno-labeling. Compared to commonly used fixation and embedding techniques, the preparation of thin sections from untreated material method s...
Espino-Solis GP, Calderon-Amador J, Calderon-Aranda ES, Licea AF, Donis-Maturano L, Flores-Romo L, Possani LD.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...
Herb VM, Url A, Tichy A, Nell B.Establishing an immunohistochemical approach for semi-quantitative assessment of the presence of immunoglobulin G (IgG) in equine, canine, and feline corneas. Methods: Healthy corneas of horses, dogs, and cats, euthanized because of a fatal disease or an unrecoverable trauma unrelated to and without a history of ophthalmic disease were formalin-fixed, paraffin-embedded, and determined to be pathomorphologically healthy by light microscopy. Automated immunohistochemistry was performed using primary antibodies against IgG, biotin-conjugated secondary antibodies, and streptavidin-peroxidase, as w...
Schultheiss PC, Collins JK, Carman J.An indirect immunoperoxidase (IP) procedure using the avidin-biotin-peroxidase complex detection technique was developed to detect viral equine herpesvirus-1 (EHV-1) antigen in formalin-fixed paraffin-embedded tissues from aborted equine fetuses. The procedure was applied to liver, lung, and other tissues from 20 cases of confirmed or suspected EHV-1-induced abortions. Specific staining was observed in tissue sections from EHV-1-infected fetuses. Positive IP staining was present in tissues of 7 cases that were also positive by fluorescent antibody (FA) and virus isolation (VI) and that had typ...
Lambertini C, Bombardi C, Zannoni A, Bernardini C, Dondi F, Morini M, Rinnovati R, Spadari A, Romagnoli N.Proteinase activated receptor 4 (PAR) in the gastrointestinal tract is involved in the regulation of inflammation and pain pathways. The aim of the present study was to evaluate the distribution and expression of PAR in the jejunum of healthy horses and in the pathologic tracts from horses undergoing surgery for herniation of the small intestine through the epiploic foramen. Eight healthy horses (Group H) and eight horses with epiploic hernia (Group EH) were included; the jejunum samples were collected at the slaughter or intraoperatively after enterectomy, respectively. To evaluate PAR expres...
Giacchi A, Marcatili M, Withers J, Knottenbelt D.A 12-year-old Warmblood mare was presented with an acute onset left hindlimb lameness associated with generalised soft tissue swelling of the entire limb and medial saphenous vein (MSV) thrombophlebitis. A presumptive diagnosis of extremity compartment syndrome (ECS) was made. Due to the clinical deterioration, emergency fasciotomy of the crural fascia and biopsy was performed. Histological and immunohistochemical examination of the samples confirmed a diagnosis of leiomyosarcoma likely originating from the tunica media of the MSV. This report is the first to describe an unique combination of ...
Herbach N, Nagel L, Zwick T, Hermanns W.Pancreatic neuroendocrine tumors of glucagon-producing cells are extremely rare in domestic animals. In this report, we describe for the first time, to our knowledge, the incidental finding of multiple glucagon-producing neuroendocrine tumors of the pancreas of a horse. The animal was euthanized due to severe local infection after tooth extraction. On postmortem examination, multiple white nodules of up to 4 cm in diameter were observed in the pancreas. Histologically, pancreatic nodules had the appearance of neuroendocrine neoplasms with positive immunoreactivity for glucagon, synaptophysin, ...
Ellerbrock RE, Podico G, Scoggin KE, Ball BA, Carossino M, Canisso IF.The expression pattern and distribution of sex steroid receptors and steroidogenic enzymes during development of the equine accessory sex glands has not previously been described. We hypothesized that equine steroidogenic enzyme and sex steroid receptor expression is dependent on reproductive status. Accessory sex glands were harvested from mature stallions, pre-pubertal colts, geldings, and fetuses. Expression of mRNA for estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2), androgen receptor (AR), 3β-Hydroxysteroid dehydrogenase/Δ5-4 isomerase (3βHSD), P450,17α hydroxylase, 17-20 lyase...
A 25-year-old pony mare was presented to the clinic with preliminarily reported severe acute colic. The pony during the previous week had shown inappetence, apathy and fever of unknown origin. Clinical examination and placement of a gastric tube were indicative of a secondary gastric dilation. Rectal exploration found moderate caecal meteorism with a tensed and painful medial taenia as well as a dilated and fluid-filled small intestine. In addition, a solid, mobile, non-painful structure of approximately 10 cm diameter was palpated ventrally. A hyperechogenic mass close to the caecum was detec...
Beccati F, Pepe M, Antinori L, Pascucci L, Chiaradia E, Mandara MT.The aim of this study was to delineate the pattern of sympathetic innervation in the suprasesamoidean region of the deep digital flexor tendon (DDFT) in horses with tendinopathy by immunohistochemical labelling for tyrosine hydroxylase (TH) and α-1 adrenergic receptor (α1-AR). Twelve forelimbs were obtained from 10 horses with DDFT tendinopathy and six feet obtained from six horses were used as healthy controls. Post-mortem radiographic, ultrasonographic and gross examinations were performed on the suprasesamoidean area of the DDFT to assess the presence of tendinopathy. Longitudinal section...
Sonea IM, Bowker RM, Broadstone RV, Robinson NE.The tone of respiratory smooth muscle is largely determined by the input from autonomic nerves. The distribution of adrenergic and selected nonadrenergic, non-cholinergic (NANC) nerves in the normal equine trachealis muscle was investigated using immunohistochemistry. The smooth muscle of the trachealis was found to contain numerous nerves immunoreactive for an enzymatic marker of adrenergic nerves, as well as many nerves immunoreactive for a putative NANC neurotransmitter, peptide histidine isoleucine, a potent bronchodilator. The tissue surrounding the respiratory smooth muscle contained num...
Jorgensen JS, Geoly FJ, Berry CR, Breuhaus BA.An 8-year-old Thoroughbred gelding was admitted for evaluation of chronic lameness of the left scapulohumeral joint of 3 months' duration. Radiography revealed a radiolucent lesion with the proximal portion of the humerus in the area of the metaphysis. Scintigraphy confirmed radiographic findings, with an increased uptake of technetium Tc 99m medronate in the proximal portion of the left humerus. A preliminary diagnosis of humeral fracture was made. Two weeks later, the horse was readmitted for clinical signs of respiratory distress. Radiographic and ultrasonographic evaluation revealed masses...
