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.
Fink C, Hembes T, Brehm R, Weigel R, Heeb C, Pfarrer C, Bergmann M, Kressin M.In the glandular stomach, gap junctional intercellular communication (GJIC) plays an important role in the gastric mucosal defense system, and loss of GJIC is associated with ulcer formation. In spite of the high incidence of gastric ulcers in horses, particularly at pars nonglandularis, the presence of gap junctions in the equine stomach has not yet been studied. The objective was to obtain basic data on the distribution of gap junction protein connexin 32 (Cx32) in the different regions of normal equine gastric mucosa. Samples of mucosa were taken from seven horses at cardiac, fundic, and py...
Arai KY, Tanaka Y, Taniyama H, Tsunoda N, Nambo Y, Nagamine N, Watanabe G, Taya K.In this study, the expression patterns of inhibins, activins, insulin-like growth factor-I (IGF-I) and steroidogenic enzymes in equine placentae recovered during the latter two-thirds of gestation were examined. Concentrations of inhibin A and inhibin pro-alphaC in endometrial and fetal placental tissue homogenates were very low during the period examined, whereas these tissues contained high concentrations of activin A. In both maternal endometrial and fetal placental tissues, activin A levels decreased as pregnancy progressed. Expression of inhibin alpha-subunit was not observed in the place...
Katkiewicz M, Witkowski L, Kita J.The aim of this study was to investigate the structure of lymphatic organs in foals which died due to Rhodococcus equi infection. The material for this study consisted of 9 foals, of both genders, various breeds, 6-14 weeks old. The routine histopathological examination and immunocytochemical reaction for cytokeratine-3 receptors revealed the disturbances in the thymus epithelial cells development. This was manifested by a complete lack of cells expressing cytokeratine-3 receptors, as well as absence of normal thymocytes differentiation. The supposed thymus epithelial cells were very few, expr...
Lepault E, Céleste C, Doré M, Martineau D, Theoret CL.Wound repair in horse limbs is often complicated by exuberant granulation tissue, a condition characterized by excessive fibroplasia and scarring and that resembles hypertrophic scars and keloids in man. The aim of this study was to compare microvascular occlusion and apoptosis in wounds of the limb with those of the body, which heal normally. Five 6.25 cm(2) wounds were created on both forelimbs and on the body of six horses. One limb was bandaged to stimulate excessive fibroplasia. Weekly biopsies were evaluated histologically and immunohistochemically for mutant p53 protein by terminal deox...
Walter I, Handler J, Miller I, Aurich C.Periglandular arrangement of myofibroblasts, associated with the deposition of extracellular matrix (ECM), is a cardinal feature of endometrosis in mares. We hypothesized that a disturbance in the expression of matrix degrading enzymes such as matrix metalloproteinases (MMP's) and matrix cross-linking proteins might lead to an imbalance in deposition and degradation of extracellular matrix components and thereby accentuate degeneration. Therefore, distributions of MMP-2, capable of collagen IV and laminin degradation, and tissue transglutaminase (TG2), a cross-linker of extracellular matrix pr...
Perillo A, Passantino G, Passantino L, Cianciotta A, Mastrosimini AM, Lacovazzi P, Venezia P, Jirillo E, Troncone A.Among 622 slaughtered horses from eastern Europe, 156 thyroid glands were selected on the basis of macroscopic alterations (e.g., determination of volume and weight). In the 80% of these thyroids, microscopic alterations consistent with a diagnosis of Hashimoto thyroiditis-like disease were found. In particular, a subverted architecture of the thyroid gland with colloid rarefaction, lymphocytic infiltration, and fibrosis was noted. The confirmation of the histopathological diagnosis of an equine Hashimoto thyroiditis-like disease was provided by the increased serum concentration of thyroglobul...
Ducharme-Desjarlais M, Céleste CJ, Lepault E, Theoret CL.To determine the effect of a silicone dressing on the rate and quality of repair of limb wounds and compare microvascular occlusion and apoptosis in wounds treated with the silicone dressing and those treated with a conventional dressing in horses. Methods: 5 horses. Methods: Horses received two 6.25-cm2 wounds on each metacarpus. Ten wounds were treated with a silicone dressing; the other 10 were treated with a control dressing. Quality of repair and wound size were evaluated at each bandage change. Time to healing and the number of excisions of exuberant granulation tissue were recorded. Bio...
Kuwano A, Ueno T, Katayama Y, Nishiyama T, Arai K.Between the laminar epidermis and the laminar dermis of laminar region (LR) in equine foot, it can be observed the basement membrane zone (BMZ), which is composed of a basement membrane and its accompaniments like the hemidesmosome and anchoring fibril. Alteration in the BMZ in equine laminitis is possibly related with not only development but also recovery outcome and recurrence of this disease. However, there is little known about the structure of the BMZ during the recovery phase of this disease. To assess the condition of the BMZ of LR affected by chronic laminitis, the tissue was examined...
Södersten F, Ekman S, Eloranta ML, Heinegård D, Dudhia J, Hultenby K.The structure and organisation of the extracellular matrix, and in particular the axial alignment of type I collagen fibrils, are essential for the tensile strength of tendons. The resident tenocytes synthesize and maintain the composition of the extracellular matrix, which changes with age and maturation. Other components of the extracellular matrix include less abundant collagen types II, III, V, VI, XII, proteoglycans and glycoproteins. Cartilage oligomeric matrix protein (COMP) is an abundant non-collagenous pentameric glycoprotein in the tendon, which can bind to collagen types I and II. ...
Hallamaa RE, Saario E, Tallberg T.Healing sarcoids were followed in 18 horses which had taken part in previous clinical studies on a total of 29 horses suffering from either primary or recurrent sarcoids, treated with bio-immunotherapy. In the present study, attention was paid to changes observed in these fibroblastic skin tumours during their regression. The tumours were surgically debulked leaving the base in the skin. The horses were immunized according to bio-immunotherapy at 2- to 4-week intervals with an autogenous vaccine made from the excised part of the tumour until the base had visibly regressed. Healing was followed...
Yamano S, Eto D, Hiraga A, Miyata H.To consider the optimal training programme for Thoroughbred horses, we examined the recruitment pattern of muscle fibres including hybrid muscle fibres in well-trained Thoroughbred horses. The horses performed exercise at three different intensities and durations; i.e., 100% VO2max for 4 min, 80% and 60% VO2max for 8 min on a treadmill with 10% incline. Muscle samples were obtained from the middle gluteal muscle before, during (4 min at 80% and 60% VO2max), and after exercise. Four muscle fibre types (types I, IIA, IIA/IIX, and IIX) were immunohistochemically identified, and optical density of...
Mirza MH, Seahorn TL, Oliver JL, Hosgood G, Moore RM.The objective of the study was to determine whether nitric oxide (NO) is present in clinically healthy horses (control) under basal conditions, and if it increases secondary to naturally acquired strangulating large colon volvulus (affected). Eleven affected horses and 10 controls were studied. Jugular venous blood, abdominal fluid, and urine were collected. The NO concentrations were standardized to the creatinine concentration in the respective samples. A biopsy specimen collected from the large colon pelvic flexure at surgery was divided into subsections for processing for inducible nitric ...
Parbhakar OP, Duke T, Townsend HG, Singh B.Horses are unique in their extreme sensitivity to endotoxin-induced cardio-pulmonary shock and mortality. The mechanisms behind increased sensitivity of the horse to endotoxin remain unknown. Pulmonary intravascular macrophages (PIMs) are pro-inflammatory cells occurring in horses. Because the functions of equine PIMs in endotoxemia remain unknown, we studied the role played by equine PIMs in endotoxin-induced pulmonary pathophysiology. We achieved this by using a recently developed protocol to deplete PIMs in order to compare lipopolysaccharide (LPS)-induced pulmonary responses in horses with...
Szeredi L, Hotzel H, Sachse K.Seventy-seven cases of equine abortion from 49 Hungarian farms that occurred between 1998 and 2000 were investigated for the presence of chlamydiae by immunohistochemistry, PCR and/or MZN staining. Evidence of the presence of these bacteria was obtained in 64 cases (83.1%) from 41 (83.7%) different farms. Partial ompA gene sequencing of PCR products revealed that the agent was Chlamydophila psittaci. Based on the findings of microbial diagnosis, pathology and case history, chlamydial infection was considered to be the most likely cause of abortion in at least 11 (14.3%) cases. In the remaining...
Kumar P, Timoney JF.The tubal tonsil of the horse surrounds the pharyngeal opening of the eustachian tube and is lined by pseudostratified columnar ciliated epithelium interspersed with areas of follicle-associated epithelium (FAE) heavily infiltrated by lymphocytes but devoid of goblet and ciliated cells. Scanning and transmission electron microscopy revealed microvillous cells and cells with features characteristic of M cells such as reduced microvilli or depressed bare surface, more numerous mitochondria, small vesicles and lysosomes, as well as vimentin filaments and epitopes specific for GS 1-B4 as previousl...
Kumar P, Timoney JF.The palatine tonsils of five young horses formed 10-12 cm elongated follicular structures extending from the root of the tongue on either side to the base of the epiglottis and lateral to the glossoepiglottic fold. The stratified squamous non-keratinized epithelium of the outer surface was modified into crypts as reticular epithelium by heavy infiltration of lymphoid cells from underlying lymphoid follicles. In places, lymphoid tissue reaching almost to the surface and with only one to two cell layers intact was identified as the lymphoepithelium. Langerhans cells with Birbeck granules were in...
Szeredi L, Hornyák A, Pálfi V, Molnár T, Glávits R, Dénes B.The occurrence of equine arteritis virus (EAV) induced equine abortions was studied with different laboratory methods during a 3-year period. Tissue samples from 96 aborted equine foetuses or newborn foals were collected from 57 farms located in different parts of Hungary. Virus isolation, polymerase chain reaction (PCR), immunohistochemistry and serology were used for the detection of EAV infection. The overall seroprevalence of EAV infection in mares was 65%. EAV induced abortion was diagnosed in eight (8.3%) cases from six (10.5%) herds. Abortion was sporadic in all herds except for one, wh...
Henson FM, Bowe EA, Davies ME.To identify the effect of fibroblastic growth factor-2 (FGF-2) on the intrinsic damage-repair response in articular cartilage in vitro. Methods: Articular equine cartilage explants, without subchondral bone, had a single impact load of 500 g applied from a height of 2.5 cm. Explants were then cultured in 0, 12, 25, 50 or 100 ng/ml FGF-2 for up to 28 days. Unimpacted discs served as controls for each time-point. Histological and immunohistochemical techniques were used to quantify and characterise the response of putative chondrocyte progenitor cells (CPC) to damage and FGF-2 treatment. Results...
Milne EM, Fintl C, Hudson NP, Pearson GT, Mayhew IG, Hahn CN.This paper describes the histopathological and immunohistochemical changes in the central, autonomic and enteric nervous systems in a well-documented case of equine dysautonomia (ED), after the animal had recovered without significant residual clinical signs. Evidence of neuronal degeneration, such as neuronal chromatolysis, glial scars or a decrease in density of neurons, was not observed in the midbrain, pons, medulla oblongata or spinal cord, including the nuclei of cranial nerves III, V, VII, X and XII. In addition, no evidence of muscle denervation or re-innervation, such as group atrophy...
Bilinska B, Hejmej A, Gancarczyk M, Sadowska J.The objective of this study was to visualize androgen receptors (ARs) in the testis, epididymis, and prostate of the stallion by means of immunohistochemistry. Nuclear immunostaining was found in all somatic cells in the testis--Leydig, Sertoli, and peritubular myoid cells; in both types of epithelial cells of the epididymis; and in the secretory cells of the prostate. These results indicate that ARs are distributed throughout the reproductive tract cells of the stallion.
Anton F, Leverkoehne I, Mundhenk L, Thoreson WB, Gruber AD.The human hCLCA1 and murine mCLCA3 (chloride channels, calcium-activated) have recently been identified as promising therapeutic targets in asthma. Recurrent airway obstruction in horses is an important animal model of human asthma. Here, we have cloned and characterized the first equine CLCA family member, eCLCA1. The 913 amino acids eCLCA1 polypeptide forms a 120-kDa transmembrane glycoprotein that is processed to an 80-kDa protein in vivo. Three single nucleotide polymorphisms were detected in the eCLCA1 coding region in 14 horses, resulting in two amino acid changes (485H/R and 490V/L). Ho...
Rüfenacht S, Marti E, von Tscharner C, Doherr MG, Forster U, Welle M, Roosje PJ.The pathogenesis of equine urticaria is not well understood. In man, urticaria has been associated with immunological and nonimmunological mechanisms leading to the release of various mediators by mast cells. Skin biopsies of 32 horses with a history of urticaria were stained with toluidine blue, a double-labelling method for chymase and tryptase, and immunohistochemistry for immunoglobulin (Ig)E. These horses were compared with horses with pemphigus foliaceus, insect bite hypersensitivity and control horses with healthy skin. Neither formalin fixation time nor biopsy site influenced the stain...
Thirion-Delalande C, Guillot J, Jensen HE, Crespeau FL, Bernex F.A 6-year-old female pony died after 2 days of prostration. Clinical signs included hyperthermia and abnormal pulmonary auscultation sounds. Necropsy revealed diffuse severe necrohaemorrhagic colitis and splenitis, multiple visceral ecchymoses, petechial haemorrhages in the brain and lungs. Microscopical examination showed acute necrohaemorrhagic colitis, encephalitis, pneumonia and splenitis associated with fibrinoid vasculitis, thrombosis and fungal hyphae within and around vessels. Immunohistologically, concomitant aspergillosis (caused by Aspergillus fumigatus) and mucormycosis (causde by A...
Hejmej A, Gorazd M, Kosiniak-Kamysz K, Wiszniewska B, Sadowska J, Bilińska B.Androgen metabolism may proceed to amplify the action of testosterone by its aromatisation to oestradiol. Recently, a growing body of evidence suggests a role of oestrogens in the male reproductive tract via their specific oestrogen receptors (ERs). In order to check whether androgens are converted to oestrogens in the testis, epididymis and prostate of the stallion, the expression of aromatase was visualised by means of immunohistochemistry. Moreover, to show the cellular targets for oestrogens the presence of oestrogen receptor alpha (ERalpha) and oestrogen receptor beta (ERbeta) was demonst...
Semevolos SA, Strassheim ML, Haupt JL, Nixon AJ.Hypertrophic differentiation and endochondral ossification of growth cartilage are regulated by a complex array of signaling peptides, including parathyroid hormone-related protein (PTH-rP), Indian hedgehog (Ihh), and bone morphogenetic proteins (BMPs). This study investigated the expression of Ihh, Patched1 and 2 (Ptc1, Ptc2), Smoothened (Smo), Gli1, and Gli3, in naturally acquired articular osteochondrosis, using an equine model. Cartilage was harvested from osteochondrosis (OC) affected femoropatellar or scapulohumeral joints from immature horses and normal control horses of similar age. Ih...
Shibahara T, Kuwano A, Ueno T, Katayama Y, Ohya T, Taharaguchi S, Yamamoto S, Umemura T, Ishikawa Y, Kadota K.Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen-containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen-containing spirochetes were found in 7 of 12 ho...
King SS, Campbell AG, Dille EA, Roser JF, Murphy LL, Jones KL.Dopamine (DA) agonist and antagonist treatments can affect ovarian reproductive events in the mare. To support our theory that DA produces these effects by acting directly on the ovary, we analyzed equine ovarian tissues for the presence of dopamine receptor-1 (D1r) and dopamine receptor-2 (D2r) mRNA by reverse transcription polymerase chain reaction (RT-PCR) and D1r and D2r proteins by Western blot and immunohistochemistry (IHC). RT-PCR was performed on RNA isolated from ovarian cortex, medulla, granulosa/theca or corpus luteum (CL) tissues and from pituitary (D2r control) and renal artery (D...
Yamano S, Eto D, Kasashima Y, Hiraga A, Sugiura T, Miyata H.To determine the growth-related changes in metabolic and anatomic properties in equine muscle fiber type, including hybrid fibers identified with immunohistochemical analysis. Methods: 24 2-, 6-, 12-, and 24-month-old female Thoroughbreds. Methods: Samples were obtained from the gluteus medius muscle of all horses. Expression of myosin heavy chain (MHC) isoforms MHC-I, -IIa, -IIb, and -IIx in each muscle fiber was detected by use of 4 primary monoclonal antibodies: BA-D5, SC-71, BF-F3, and BF-35, respectively. Five muscle fiber types (types I, I/IIA, IIA, IIA/IIX, and IIX) were immunohistochem...
Nixon C, Chambers G, Ellsmore V, Campo MS, Burr P, Argyle DJ, Reid SW, Nasir L.Equine sarcoids are benign fibroblastic skin tumours affecting equids worldwide. Whilst the pathogenesis is not entirely understood, infection with Bovine Papillomavirus (BPV) types 1 and 2 has been implicated as a major factor in the disease process, however the mechanism by which BPV infection contributes to sarcoid pathology is not clear. In this study, we show that the majority of sarcoids express the BPV-1 major transforming gene E6. Further, we demonstrate that sarcoid lesions are not associated with high levels of cellular proliferation as assessed by Ki67 expression or with expression ...
McFarlane D, Dybdal N, Donaldson MT, Miller L, Cribb AE.Equine pituitary pars intermedia dysfunction (PPID) is a spontaneously occurring progressive disease affecting aged horses and ponies. The pathogenesis of PPID is poorly understood, but the available evidence supports a loss of dopaminergic inhibition of the melanotropes of the pars intermedia. Horses with PPID have increased plasma concentrations of pars intermedia pro-opiomelanocortin-derived peptides that decrease in response to dopamine or dopamine agonist administration. Dopamine and dopamine metabolite concentrations are decreased in the pars intermedia of affected horses compared to age...
Ulrich R, Eydner M, Grün A, Haydn J, Baumgärtner W.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...
Yamashita-Kawanishi N, Ito S, Chambers JK, Uchida K, Sato M, Chang HW, Knight C, van der Meer F, Haga T.Equus caballus papillomavirus type 2 (EcPV2) infection has been associated with genital squamous cell carcinoma (SCC) development in horses. However, very few reports on EcPV2-associated disease in Asia exist. Our study characterizes pathological and virological features of an EcPV2-associated vulvar SCC from a Japanese mare. Conventional PCR, in situ hybridization, reverse-transcriptase PCR and immunohistochemistry confirmed the presence and distribution of EcPV2 within the lesion and suggested that p53 degradation may not be the mechanism by which this virus induces neoplastic transformation...
Lindner A, Dag S, Marti-Korff S, Quiroz-Rothe E, López Rivero JL, Drommer W.The collection of muscle samples in horses to estimate their performance potential, adaptations to training and diagnosis of exertional myopathies is of increasing interest. This study was designed to examine whether repeated biopsying of the gluteus medius muscle at 7 week intervals alters morphology and muscle fibre variables in the sampling area. The gluteus had been biopsied 3 times previously, each biopsy separated by a 7 week interval. Seven weeks after the last biopsy, this study was initiated. Percutaneous needle biopsies were taken from left and right muscles at 2 depths (2 and 6 cm) ...
Weaver RE, Sharif M, Livingston LA, Andrews KL, Fuller CJ.The objective of this study was to assess whether macroscopically normal articular cartilage taken from joints containing focal osteoarthritic lesions is histologically similar to articular cartilage taken from macroscopically normal joints. Metacarpophalangeal, proximal interphalangeal, and distal interphalangeal joints were obtained from 10 horses following euthanasia. Gross articular cartilage damage was scored and the cartilage assigned to one of two groups: (1) macroscopically normal cartilage from normal joints (control) and (2) macroscopically normal cartilage from diseased joints in wh...
Townsend J, Sneddon CL, Tortonese DJ.The intrapituitary mechanisms regulating gonadotrophin secretion in the horse remain unclear. Here, we examined seasonal and gonadal effects on the gonadotroph and lactotroph populations of male horses with the aim of defining a possible morphological basis for the differential release of gonadotrophins. Pituitaries were collected from: (i) gonadal-intact horses in the breeding season (GBS); (ii) orchidectomized horses in the breeding season (OBS); and (iii) orchidectomized horses in the nonbreeding season (ONBS). Immunohistochemistry was performed using antibodies to the luteinizing hormone (...
Porter BF, Storts RW, Payne HR, Edwards JF.Ganglioneuromas are complex tumors that arise in peripheral ganglia and are composed of well-differentiated neurons, nerve processes, Schwann cells, and enteric glial cells. The term ganglioneuromatosis (GN) denotes a regional or segmental proliferation of ganglioneuromatous tissue. This report describes an 8-year-old mixed breed horse with GN in a 25-cm segment of small colon. Grossly, the lesion consisted of numerous sessile to pedunculated nodules extending from the serosal surface. Histologic examination revealed the nodules to consist of fascicles of spindle-shaped cells consistent with S...
Ricard RM, Burton J, Chow-Lockerbie B, Wobeser B.Chlamydiae are reported to cause abortion in several species, however the association between Chlamydia sp. and equine abortions is poorly understood. A zoonotic transfer event of C. psittaci from aborted equine tissues in Australia has emphasized the need to better understand the prevalence of this pathogen in equine populations. The prevalence of chlamydia in equine abortions in North America has not been investigated thoroughly. We examined 99 formalin-fixed, paraffin-embedded placental samples submitted between 2009 and 2020 from equine abortions in Western Canada using chlamydia-specific ...
Newkirk KM, Ehrensing G, Odoi A, Boston RC, Frank N.OBJECTIVE To assess insulin, glucagon, and somatostatin expression within pancreatic islets of horses with and without insulin resistance. ANIMALS 10 insulin-resistant horses and 13 insulin-sensitive horses. PROCEDURES For each horse, food was withheld for at least 10 hours before a blood sample was collected for determination of serum insulin concentration. Horses with a serum insulin concentration 20 μU/mL underwent a frequently sampled IV glucose tolerance test to determine sensitivity to insulin by minimal model analysis. Horses with a sensitivity to insulin < 1.0 × 10 L•min•mU w...
LeRoy BE, Knight MC, Eggleston R, Torres-Velez F, Harmon BG.A 14-year-old bay Thoroughbred gelding was presented for evaluation of a mass at the base of the tail. The mass had been present for 1 year, and recently had begun to increase in size. Additional masses were found around the eye and shoulder. A fine-needle aspirate of the tail-base mass revealed highly anaplastic round to polyhedral cells containing dark green to black cytoplasmic granules interpreted to be melanin. Histologically, the mass was composed of pleomorphic, poorly pigmented, round to polyhedral cells interpreted to be neoplastic melanocytes. With immunohistochemistry, the cells wer...
Lisowski ZM, Sauter KA, Waddell LA, Hume DA, Pirie RS, Hudson NPH.Intestinal macrophages are the largest group of mononuclear phagocytes in the body and play a role in intestinal innate immunity, neuroimmune interactions and maintaining intestinal homeostasis. Conversely, they also are implicated in numerous pathologies of the gastrointestinal tract, such as postoperative ileus and inflammatory bowel disease. As a result, macrophages could be potential therapeutic targets. To date, there are limited studies on the morphology and distribution of macrophages in the equine gastrointestinal tract (GIT). The aim of this study was to identify the location and abun...
Jago RC, Scholes S, Mair TS, Pearson GR, Pirie RS, Handel I, Milne EM, Coyle F, Mcgorum BC.An accurate, minimally invasive, ante-mortem diagnostic test for equine grass sickness (EGS) is currently lacking. Although histological examination of haematoxylin and eosin-stained rectal biopsies for chromatolytic neurons is insensitive as a diagnostic test for EGS, we hypothesised that its diagnostic accuracy could be improved by immunolabelling for β-amyloid precursor protein (β-APP), which has increased expression in cranial cervical ganglia (CCG) neuronal perikarya in EGS. Objective: To develop a grading scheme for assessing the distribution and intensity of β-APP immunoreactivity wi...
Ben Hamouda S, Vargas A, Boivin R, Miglino MA, da Palma RK, Lavoie JP.Severe asthma is associated with an increased airway smooth muscle (ASM) mass and altered composition of the extracellular matrix (ECM). Studies have indicated that ECM-ASM cell interactions contribute to this remodeling and its limited reversibility with current therapy. Three-dimensional matrices allow the study of complex cellular responses to different stimuli in an almost natural environment. Our goal was to obtain acellular bronchial matrices and then develop a recellularization protocol with ASM cells. We studied equine bronchi as horses spontaneously develop a human asthma-like disease...
Millares-Ramirez EM, Lavoie JP.Severe asthma in horses is characterized by structural changes that thicken the lower airway wall, a change that is only partially reversible by current treatments. Increased vascularization contributes to the thickening of the bronchial wall in humans with asthma and is considered a potential new therapeutic target. Objective: To determine the presence of angiogenesis in the bronchi of severely asthmatic horses, and if present, to evaluate its reversibility by treatment with corticosteroids. Methods: Study 1: Bronchial samples from asthmatic horses in exacerbation (7), in remission (7), and a...
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 ...
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 ...
Kydd JH, Butcher GW, Antczak DF, Allen WR.The expression of class I Major Histocompatibility Complex (MHC) molecules by early trophoblast of intraspecific horse and donkey, interspecific mule and extraspecific donkey-in-horse conceptuses was determined using a rat monoclonal antibody (MAC 291) in a peroxidase anti-peroxidase immunohistochemical technique. Most non-invasive allantochorion of horse, donkey and mule conceptuses did not express class I MHC molecules at any stage of gestation except in small isolated patches of pseudostratified trophoblast lying adjacent to the openings of endometrial glands. In contrast, MHC class I molec...
Katayama Y, Wada R, Kanemaru T, Sasagawa T, Uchiyama T, Matsumura T, Anzai T.Equine protozoal myeloencephalitis developed in a three-year-old male Thoroughbred racehorse imported from the United States. The animal showed astasia five days after the onset of ataxia. Histopathologically, focal nonpurulent myelitis accompanied by hemorrhage and perivascular infiltration was observed in the fourth and fifth cervical spinal cord. Immunohistochemically, shizonts were occasionally observed and were positive for anti-Sarcocystis neurona (S. neurona) antiserum. S. neurona-specific antibodies were detected in the serum and cerebrospinal fluid by Western blot. This is the first e...
ERas is a new gene of the Ras family found in murine embryonic stem (ES) cells. Its human ortholog is not expressed in human ES cells. So far ERas gene has only been found to be expressed in the tissues of adult cynomolgus monkeys and cattle; however, information about ERAS expression or its potential functions in equine tissues is lacking. This study was performed to investigate whether Eras is an equine functional gene and whether ERAS is expressed in the tissues of adult horses and determine its potential physiological role. Expression of the ERas gene was detected in all examined adult tis...
Arrighi S, Domeneghini C.An immunohistochemical study was carried out on specimens of testis, excurrent duct including the male accessory glands and urethra in its various tracts in the horse and the donkey, in order to localize nine regulatory peptides. Immunoreactivities were tested by means of Labelled Strept Avidin-Biotin (LSAB) method. The study has shown that Equine male genitalia are supplied by many peptide immunoreactive nerves containing NPY-, VIP-, leu- and met-Enkephalin-, Substance P-, CGRP- and Bombesin/GRP-like peptides, each of them having a characteristic distribution pattern. These neurotransmitters ...
Weng Q, Tanaka Y, Taniyama H, Tsunoda N, Nambo Y, Watanabe G, Taya K.To elucidate the relationship between steroidogenic hormones and developing adrenal glands, we investigated the immunolocalization of steroidogenic enzymes in equine fetal adrenal glands during mid-late gestation. Fetal adrenal glands were obtained from three horses at 217, 225 and 235 days of gestation. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17) and huma...
Meichner K, Kraszeski BH, Durrant JR, Grindem CB, Breuhaus BA, Moore PF, Neel JA, Linder KE, Borst LB, Fogle JE, Tarigo JL.An 11-year-old, 443-kg Haflinger mare was presented to the North Carolina State University Veterinary Teaching Hospital with a 2-week history of lethargy and a 3-day duration of anorexia, pyrexia, tachycardia, and ventral edema. Severe pitting edema, peripheral lymphadenopathy, and a caudal abdominal mass were noted on physical examination. An extreme leukocytosis (154.3 × 103 /μL) and microscopic hematologic findings suggestive of myelomonocytic leukemia were observed. Serum protein electrophoresis revealed a monoclonal gammopathy and urine protein electrophoresis revealed a monoclonal ligh...
Lee KH, Park C, Bang H, Ahn M, Moon C, Kim S, Shin T.The olfactory mucosae of the horse were examined by using histology and lectin histochemistry to characterize the carbohydrate sugar residues therein. Histological findings revealed that olfactory epithelium (OE) consisted of both olfactory marker protein (OMP)- and protein gene product (PGP) 9.5-positive receptor cells, supporting cells and basal cells with intervening secretory ducts from Bowman's glands. Mucus histochemistry showed that Bowman's gland acini contain periodic acid-Schiff (PAS) reagent-positive neutral mucins and alcian blue pH 2.5-positive mucosubstances. Lectin histochemistr...
Weijs W, Dingboom E, van Ginkel F.The contribution to total variance of different error sources in fibre type counts of equine gluteus medius muscle biopsies was determined to quantify and possibly improve the resolution of the method. Fibre types were defined on the basis of myosin heavy chain immunostaining. Errors were determined at levels: (1) positioning the insertion channel, (2) positioning the needle tip (3) biopsy heterogeneity (4) observer interpretation. Errors at levels 1 and 2 were considerable. Confidence intervals for individual observations were +/- 10-15%. In longitudinal studies a group size of 4 animals is n...
Bengtsdotter EA, Ekman S, Andersen PH.Inguinal pain, unexplained hind limb lameness, back pain or behavioural problems in geldings could be attributable to painful neuromas that develop as a consequence of crushing and severing the testicular nerves during castration. The presence of neuroma in this anatomical location has never been reported, hence the knowledge of possible clinical relevance is limited. The aim of this study was to histologically investigate the testicular nerves at the castration site in geldings for the presence of neuromas. Proximal spermatic cord remnants were collected from 20 geldings admitted to routine p...
Torquato RBC, Iamamoto K, Fernandes ER, Achkar S, Silva SR, Katz ISS, Guedes F.Laboratory diagnosis of rabies in equines is essential for distinguishing the disease from other sources of encephalitis. Diagnosis by conventional techniques such as a direct fluorescent antibody test (dFAT) or viral isolation in mice or cell culture can be difficult, and the application of molecular biological methods may be necessary. We performed an indirect rapid immunohistochemistry test (iRIT) for the detection of the rabies virus (RABV) antigen in the central nervous system (CNS) of equines and compared the results with those of other diagnostic techniques. We reviewed result records f...
Kagawa Y, Hirayama K, Tagami M, Tsunoda N, Yoshino T, Matsui T, Furuoka H, Taniyama H.Histopathological and immunohistochemical examinations were made on four female horses aged 9-12 years with pulmonary granular cell tumours (GCTs). The tumours, which were multiple, of varying size, firm and off-white in colour, surrounded the bronchi and bronchioles. Metastatic lesions were not detected. The tumour cells had abundant eosinophilic cytoplasm filled with prominent coarse eosinophilic granules. Immunohistochemically, these tumour cells reacted uniformly with vimentin and S100 antibodies. Most were immunolabelled by antibodies against glial fibrillary acidic protein (GFAP), myelin...
Yoon M, Jiang J, Chung KH, Roser JF.Insulin-like growth factor plays a paracrine/autocrine role in regulating testicular function in the stallion, but its presence in the equine epididymis remains unknown. The aim of this study was to test the hypothesis that insulin-like growth factor-I (IGF-I) and IGF-I receptor (IGF-IR) are localized in the caput, corpus, and cauda of the epididymis in an age-dependent manner. Immediately after castration, epididymal tissue was fixed, paraffin-embedded, and processed for immunohistochemistry (IHC). Western blot was also performed using equine epididymal extracts to verify the specificity of t...
Welter H, Bollwein H, Weber F, Rohr S, Einspanier R.The expression of the endothelial and inducible nitric oxide synthases (eNOS and iNOS, respectively) was examined in the endometrium of cyclic and pregnant mares by real-time polymerase chain reaction and immunohistology. The concentration of eNOS mRNA varied throughout the oestrous cycle, with significantly higher transcripts on Day 5 of the oestrous cycle (P 0.05). In early pregnant mares both eNOS and iNOS mRNA increased between Days 12 and 15 (P < 0.05). In cyclic mares, eNOS protein was detected immunocytochemically in endometrial epithelia, the basement membrane, the endothelial laye...
Szeredi L, Aupperle H, Steiger K.Formalin-fixed, paraffin wax-embedded fetal membranes from 76 cases of equine abortion were examined immunohistochemically for equine herpesvirus (EHV)-1 antigen. Of the 76 cases, 11 had been proved EHV-1-positive by diagnostic methods applied to the aborted fetuses (viral isolation in tissue culture, or immunohistochemical examination, or both). Of the 11 fetal membranes from the virus-positive animals, five gave positive results on immunohistochemical examination, and three on in-situ hybridization; the positive signals were detected in trophoblastic cells and occasionally in monocytes and e...
Hoque S, Derar RI, Senba H, Osawa T, Kano K, Taya K, Miyake Y.To clarify the morphological and immunohistochemical characteristics in mares with granulosa theca cell tumor (GTCT), the localization of inhibin subunits (alpha, betaA, betaB) and aromatase in the granulosa cell layers and theca layers in the ovarian follicles were determined by immunohistochemical staining. The follicles were obtained from the ovaries of 6 mares with GTCT and 4 normal mares as controls. Immunohistochemically, inhibin alpha-subunit was localized in the granulosa cells of all follicles showing different sizes in all GTCT cases and betaA- subunit was localized in two GTCT cases...
