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.
Sampieri F, Allen AL, Alcorn J, Clark CR, Vannucci FA, Pusterla N, Mapes SM, Ball KR, Dowling PM, Thompson J, Bernstein LR, Gebhart CJ, Hamilton DL.Antimicrobial efficacy against Lawsonia intracellularis is difficult to evaluate in vitro, thus, the effects of gallium maltolate's (GaM) were investigated in a rabbit model for equine proliferative enteropathy (EPE). Juvenile (5-6-week-old) does were infected with 3.0 × 10(8) L. intracellularis/rabbit and allocated into three groups (n = 8). One week postinfection, one group was treated with GaM, 50 mg/kg; one, with doxycycline, 5 mg/kg; and one with a sham-treatment (control). Feces and blood were collected daily and weekly, respectively, to verify presence of L. intracellularis fec...
Dürrwald R, Kolodziejek J, Weissenböck H, Nowotny N.Borna disease (BD) is a sporadic neurologic disease of horses and sheep caused by mammalian Borna disease virus (BDV). Its unique epidemiological features include: limited occurrence in certain endemic regions of central Europe, yearly varying disease peaks, and a seasonal pattern with higher disease frequencies in spring and a disease nadir in autumn. It is most probably not directly transmitted between horses and sheep. All these features led to the assumption that an indigenous virus reservoir of BDV other than horses and sheep may exist. The search for such a reservoir had been unsuccessfu...
Gauff FC, Patan-Zugaj B, Licka TF.To determine the effect of short-term hyperinsulinemia on the localization and expression of endothelin receptor (ETR)-A and ETR-B in lamellar tissue of the forelimbs of horses. Methods: Distal portion of 15 cadaveric forelimbs from healthy adult horses (1 limb/horse) obtained immediately after slaughter at an abattoir. Methods: Each forelimb was assigned to 1 of 3 treatment groups (perfused with autologous blood for 10 hours [control perfusion; n = 5], perfused with an insulin [142 ± 81 μU/mL] perfusate for 10 hours [insulinemic perfusion; 5], or not perfused [unperfused control; 5]). Immun...
Jung HJ, Song H, Yoon MJ.Deleted in azoospermia-like (DAZL) is used as a germ cell marker in several species, including mice, rats, pigs, rhesus monkeys, bulls, and humans. Our objectives with this study were to investigate DAZL expression in stallion germ cells by using immunofluorescence, immunocytochemistry, and western blotting, and to determine the effects of reproductive stage and breeding season on the DAZL-positive cell population in seminiferous tubule cross sections. Testes were obtained during routine castration procedures at a large animal clinic and routine field service castration. The reproductive stage...
Cadby JA, Buehler E, Godbout C, van Weeren PR, Snedeker JG.The role of intrinsic and extrinsic healing in injured tendons is still debated. In this study, we characterized cell plasticity, proliferative capacity, and migration characteristics as proxy measures of healing potential in cells derived from the peritenon (extrinsic healing) and compared these to cells from the tendon core (intrinsic healing). Both cell populations were extracted from horse superficial digital flexor tendon and characterized for tenogenic and matrix remodeling markers as well as for rates of migration and replication. Furthermore, colony-forming unit assays, multipotency as...
Zannoni A, Bombardi C, Dondi F, Morini M, Forni M, Chiocchetti R, Spadari A, Romagnoli N.Proteinase-activated receptor 2 (PAR2) is a G-protein-coupled receptor for trypsin and mast cell tryptase; it is highly expressed at the intestinal level with multiple functions, such as epithelial permeability and intestinal motility. Many proteases activate PAR2 during tissue damage, suggesting a role of the inflammatory response receptors. The aim of the study was to evaluate the distribution and expression of PAR2 in the jejunum, the ileum and the pelvic flexure, using samples collected from healthy adult horses after slaughter. Proteinase-activated receptor 2 immunoreactivity (PAR2-IR) wa...
Arkoun B, Gautier C, Delalande C, Barrier-Battut I, Guénon I, Goux D, Bouraïma-Lelong H.Among mammals, the stallion produces the largest amount of testicular estrogens. These steroid hormones are produced mainly by Leydig and Sertoli cells in the testis and also in the epididymis. Their role in horse testicular physiology and their ability to act on spermatozoa are still unknown. In order to determine if spermatozoa are targets for estrogens, the presence of estrogen receptors in mature ejaculated spermatozoa has been investigated. The presence of a single isoform of ESR1 (66kDa) and ESR2 (61kDa) was found by Western-blot analysis in samples from seven stallions. Confocal analysi...
Balaratnasingam C, Kang MH, Yu P, Chan G, Morgan WH, Cringle SJ, Yu DY.Retinal ganglion cell (RGC) axonal structure and function in the optic nerve head (ONH) is predominantly supported by astrocytes and capillaries. There is good experimental evidence to demonstrate that RGC axons are perturbed in a non-uniform manner following ONH injury and it is likely that the pattern of RGC axonal modification bears some correlation with the quantitative properties of astrocytes and capillaries within laminar compartments. Although there have been some excellent topographic studies concerning glial and microvascular networks in the ONH our knowledge regarding the quantitati...
Power J, Hernandez P, Wardale J, Henson FM.Osteochondrosis (OC) is a common and clinically important joint disease that occurs in many species, including humans, pigs, chickens and horses. It has been described as a focal failure of endochondral ossification (EO), but no cellular/molecular mechanisms are fully described that explain the cause of this condition. Recently a Wnt signalling inhibitor, sclerostin, has been described in osteoarthritic cartilage, where it has been proposed to protect damaged cartilage from degradation. Cartilage degradation is a key event in EO, thus, abnormalities of sclerostin in growth cartilage could, pot...
Svala E, Thorfve AI, Ley C, Henriksson HK, Synnergren JM, Lindahl AH, Ekman S, Skiöldebrand ES.To determine the effects of interleukin (IL)-6 and IL-1β stimulation on expression of growth differentiation factor (GDF)-5 and Wnt signaling pathway genes in equine chondrocytes. Methods: Macroscopically normal articular cartilage samples from 6 horses and osteochondral fragments (OCFs) from 3 horses. Methods: Chondrocyte pellets were prepared and cultured without stimulation or following stimulation with IL-6 or IL-1β for 1, 2, 12, and 48 hours; expression of GDF-5 was determined with a quantitative real-time PCR assay. Expression of genes in various signaling pathways was determined with ...
de Brot S, Junge H, Hilbe M.Two horses were presented with non-specific clinical signs of several weeks' duration and were humanely destroyed due to a poor prognosis. At necropsy examination, both horses had multiple small, white nodules replacing pancreatic tissue and involving the serosal surface of the abdominal cavity, the liver and the lung. Microscopically, neoplastic cells were organized in acini and contained abundant (case 1) or sparse (horse 2) intracytoplasmic zymogen granules. Immunohistochemically, both tumours expressed amylase and pan-cytokeratin, but not insulin or neuron-specific enolase. In case 2, a lo...
Mohanty N, Gulati BR, Kumar R, Gera S, Kumar P, Somasundaram RK, Kumar S.Mesenchymal stem cells (MSCs) isolated from umbilical cord blood (UCB) in equines have not been well characterized with respect to the expression of pluripotency and mesenchymal markers and for tenogenic differentiation potential in vitro. The plastic adherent fibroblast-like cells isolated from 13 out of 20 UCB samples could proliferate till passage 20. The cells expressed pluripotency markers (OCT4, NANOG, and SOX2) and MSC surface markers (CD90, CD73, and CD105) by RT-PCR, but did not express CD34, CD45, and CD14. On immunocytochemistry, the isolated cells showed expression of CD90 and CD73...
Dohmen PM, da Costa F, Lopes SV, Vilani R, Bloch O, Konertz W.In the past, successful use of decellularized xenogenic tissue was shown in the pulmonary circulation. This study, however, evaluates a newly developed decellularized equine pericardial patch under high pressure circumstances. Methods: Seven decellularized equine pericardial scaffolds were implanted into the descending aorta of the juvenile sheep. The implanted patches were oversized to evaluate the durability of the decellularized tissue under high surface tension (Law of Laplace). After 4 months of implantation, all decellularized patches were inspected by gross examination, light microscopy...
Kegler K, Mundle K, Walliser U, Wohlsein P.Schwannomas arising in the parotid gland are rare in man and are not documented in domestic animals. This report describes the clinical, morphological and immunohistochemical findings of a benign schwannoma in the parotid gland of a 12-year-old warmblood gelding. A slow-growing mass was surgically excised from the parotid gland and did not recur within the following 6 months. The tumour was well circumscribed and was composed of densely packed spindle cells partly arranged in an Antoni A pattern and intermixed with hypocellular areas resembling the Antoni B pattern. Tumour cells expressed vime...
van den Top JG, Harkema L, Ensink JM, Barneveld A, Martens A, van de Lest CH, van Weeren PR, Gröne A.Penile and preputial papilloma and squamous cell carcinoma (SCC) are commonly diagnosed in horses. Papillomas have the potential to progress to potentially lethal SCC. Knowledge of pathogenetic mechanisms may help in prevention and definition of treatment targets. Methods: Retrospective study using archived material. Objective: To determine the expression of cyclo-oxygenase 1 (COX-1), cyclo-oxygenase 2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) in penile and preputial normal tissue, papilloma and SCC in horses, and whether expression of these enzymes is influenced by degree of...
Pasquel SG, Agnew D, Nelson N, Kruger JM, Sonea I, Schott HC.Although the equine renal pelvis and terminal recesses have been described post mortem, little information exists about the endoscopic appearance of these structures in the living horse for guiding ureteropyeloscopy. Objective: To further document the anatomy of the upper urinary collecting system, specifically the renal pelvis and terminal recesses, of the horse. Methods: Descriptive study of cadaver material. Methods: Kidneys were harvested from 10 horses. Magnetic resonance imaging was performed after distension of the renal pelvis with an elastomer casting material, followed by visual insp...
Bolfa P, Nolf M, Cadoré JL, Catoi C, Archer F, Dolmazon C, Mornex JF, Leroux C.EIA (Equine Infectious Anemia) is a blood-borne disease primarily transmitted by haematophagous insects or needle punctures. Other routes of transmission have been poorly explored. We evaluated the potential of EIAV (Equine Infectious Anemia Virus) to induce pulmonary lesions in naturally infected equids. Lungs from 77 EIAV seropositive horses have been collected in Romania and France. Three types of lesions have been scored on paraffin-embedded lungs: lymphocyte infiltration, bronchiolar inflammation, and thickness of the alveolar septa. Expression of the p26 EIAV capsid (CA) protein has been...
Hudson NP, Pearson GT, Mayhew IG, Proudman CJ, Burden FA, Fintl C.Intestinal motility disorders are an important problem in horses and donkeys and this study was carried out in order to evaluate the enteric neurons in animals with and without intestinal disease. Surplus intestinal tissue samples were collected from 28 horses undergoing exploratory laparotomy for colic. In addition, surplus intestinal samples from 17 control horses were collected immediately following humane destruction for clinical conditions not relating to the intestinal tract. Similar samples were also collected during routine post-mortem examinations from 12 aged donkeys; six animals wer...
Pérez-Écija A, Estepa JC, Barranco I, Rodríguez-Gómez IM, Mendoza FJ, Gómez-Laguna J.Verrucous hemangiomas are a rare specific variant of equine skin tumors not well described in the literature. An 8-year-old gelding presented a unilateral lesion on the pastern. Macroscopically, the mass showed a warty and verrucous surface with focal ulcerations. The histology showed a dermal proliferation of endothelial-layered capillaries and venules separated by a delicate stroma of scant fibroblasts and collagen deposition, with pseudoepitheliomatous hyperplasia (exuberant reactive irregular epithelial hyperplasia with tongue-like projections extending into the dermis, mimicking downgrowt...
Pearl CA, Roser JF.Lactoferrin is one of the most abundant proteins secreted by the stallion epididymis, but its cellular localization and regulation remain unknown. This study was designed to address the following objectives: (1) identify the epididymal cell types producing lactoferrin in pre-pubertal, peri-pubertal and post-pubertal animals; (2) demonstrate that lactoferrin binds to stallion sperm; and (3) determine if testosterone and estradiol regulate lactoferrin secretion in vitro. Using an immunohistochemical method, lactoferrin was localized in the cytoplasm of principal cells in the corpus and cauda of ...
de Linde Henriksen M, Andersen PH, Mietelka K, Farina L, Thomsen PD, Plummer CE, Mangan BG, Heegaard S, Coleman JK, Toft N, Brooks DE.To investigate histopathologic and immunohistochemical aspects of equine deep stromal abscesses (DSA) with a focus on the histopathologic diagnosis, presumptive etiology, and the immunohistochemical expression of three angiogenesis-related factors: vascular endothelial growth factor-A (VEGF-A), pigment epithelium-derived factor (PEDF), and interleukin-1 receptor antagonist (IL-1ra). Methods: Paraffin-embedded biopsy samples from 51 DSA. The biopsies were collected from full-thickness penetrating keratoplasty or split-thickness lamellar keratoplasty surgeries at the University of Florida Veteri...
Herman EH, Knapton A, Liu Y, Lipshultz SE, Estis J, Todd J, Woodward RA, Cochran T, Zhang J, Poirier MC.Cardiac troponins serve as serum biomarkers of myocardial injury. The current study examined the influence of age on serum concentrations of cardiac troponin I (cTnI). An ultrasensitive immunoassay was used to monitor cTnI concentrations in Sprague-Dawley (SD) rats and Erythrocebus patas monkeys of different ages. The mean cTnI concentrations were highest in 10-day-old rats compared to 25-, 40-, and 80-day-old SD rats. Cardiomyocyte remodeling was apparent in hearts from 10-day-old SD rats as evident by hypercellularity, irregularly shaped nuclei, and moderate numbers of myocytes undergoing mi...
Williams KJ, Robinson NE, Lim A, Brandenberger C, Maes R, Behan A, Bolin SR.Gammaherpesviruses (γHV) are implicated in the pathogenesis of pulmonary fibrosis in humans and murine models of lung fibrosis, however there is little direct experimental evidence that such viruses induce lung fibrosis in the natural host. The equine γHV EHV 5 is associated with equine multinodular pulmonary fibrosis (EMPF), a progressive fibrosing lung disease in its natural host, the horse. Experimental reproduction of EMPF has not been attempted to date. We hypothesized that inoculation of EHV 5 isolated from cases of EMPF into the lungs of clinically normal horses would induce lung fibr...
Szóstek AZ, Lukasik K, Galvão AM, Ferreira-Dias GM, Skarzynski DJ.The aim of the study was to characterize endometrial mRNA transcription, immunolocalization, and protein expression of interleukin (IL) 1alpha, IL1beta, IL6, and IL1RI, IL1RII, and IL6Ralpha/beta in the course of endometrosis during the estrous cycle. Additionally, the influence of IL1alpha, IL1beta, and IL6 on prostaglandin (PG) secretion and PG synthase mRNA transcription in endometrial tissue during endometrosis was investigated. The endometrial samples were obtained at the early (n = 12), mid- (n = 12), and late (n = 12) luteal phases and at the follicular (n = 12) phase of the estrous cyc...
Sampieri F, Allen AL, Pusterla N, Vannucci FA, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ.The objective of this study was to demonstrate the susceptibility of rabbits to Lawsonia intracellularis obtained from a case of clinical equine proliferative enteropathy (EPE). This is a preliminary step toward developing a rabbit infection model for studying pathogenesis and therapy of EPE in horses. Nine does were equally assigned to 3 groups. Animals in 2 groups (Group 1 and Group 2) were orally inoculated with different doses of cell-cultured L. intracellularis. Controls (Group 3) were sham-inoculated. Feces and blood were collected before the rabbits were infected and at 7, 14, and 21 da...
Schnabel CL, Wagner S, Wagner B, Durán MC, Babasyan S, Nolte I, Pfarrer C, Feige K, Murua Escobar H, Cavalleri JM.Research on equine cytokines is often performed by analyses of mRNA. For many equine cytokines an analysis on the actual protein level is limited by the availability of antibodies against the targeted cytokines. Generation of new antibodies is ongoing but time consuming. Thus, testing the reactivity of commercially available antibodies for cross-reactivity with equine cytokines is of particular interest. Fifteen monoclonal antibodies against IL-1β, IL-6, IL-8, IL-12, IL-18 and Granulocyte Macrophage Colony stimulating factor (GM-CSF) of different species were evaluated for reactivity with the...
Theoret CL, Olutoye OO, Parnell LK, Hicks J.To compare histopathologic features of a fibroproliferative disorder in horses (exuberant granulation tissue-EGT) and people (keloid). Methods: Archival tissue samples of EGT (n = 8) and keloid (12). Methods: After automated hematoxylin and eosin, histochemical (Gomori trichrome, Verhoeff-van Gieson elastin) and immunohistochemical (vimentin, α-smooth muscle actin, CD34, CD68, CD117) stainings, tissue sections were evaluated using a semi-quantitative grading scale for presence or absence of ulceration, keloidal collagen, myofibroblasts, and elastic fibers as well as degree of inflammation, fi...
Stack A, Derksen FJ, Sordillo LM, Williams KJ, Stick JA, Brandenberger C, Steibel JP, Robinson NE.To determine the effects of 2 weeks of intense exercise on expression of markers of pulmonary venous remodeling in the caudodorsal and cranioventral regions of the lungs of horses. Methods: 6 horses. Methods: Tissue samples of the caudodorsal and cranioventral regions of lungs were obtained before and after conditioning and 2 weeks of intense exercise. Pulmonary veins were isolated, and a quantitative real-time PCR assay was used to determine mRNA expression of matrix metalloproteinase-2 and -9, tissue inhibitor of metalloproteinase-1 and -2, collagen type I, tenascin-C, endothelin-1, platelet...
Campos-Junior PH, Costa GM, de Avelar GF, Segatelli TM, Lacerda SM, Aponte PM, de França LR.Morphometry is a classical quantitative method often used in biology to provide a data basis for functional interpretations/interactions of a particular organ or system. Herein we took advantage of this valuable approach to evaluate the spermatogonial stem cell niche using the horse testis and immunocytochemical localization of GFRA1 [glial cell line-derived neurotrophic factor receptor produced by Sertoli cells)] as an example. Using the NIH ImageJ free software, we describe in detail all the necessary steps to investigate this specific and crucial microenvironment. Based on several recently ...
Zamboulis DE, Senior M, Clegg PD, Milner PI.Tissue sensitisation and chronic pain have been described in chronic-active laminitis in the horse, making treatment of such cases difficult. Purinergic P2X receptors are linked to chronic pain and inflammation. The aim of this study was to examine the expression of purinergic P2X receptor subtypes 1, 2, 3 and 7 in the hoof, palmar digital vessels and nerve, dorsal root ganglia and spinal cord in horses with chronic-active laminitis (n=5) compared to non-laminitic horses (n=5). Immunohistochemical analysis was performed on tissue sections using antibodies against P2X receptor subtypes 1-3 and ...
Huang L, Palmieri C, Bertin FR.Pituitary pars intermedia dysfunction (PPID) is an endocrinopathy commonly affecting old horses. It is a spontaneously occurring, progressive disease that is still poorly understood. Previous studies have observed neurodegeneration of the dopaminergic inhibition of melanotrophs, which leads to decreased dopamine (DA) in the pars intermedia (PI) and increased pro-opiomelanocortin-derived peptides circulating in plasma. However, rats knockout for the dopamine D2 receptor (D2r) similarly develop PI hypertrophy and hyperplasia. Thus, based on the current pathophysiological theory of PPID, whether ...
Swadzba ME, Hirmer S, Amann B, Hauck SM, Deeg CA.Although the presence of IgG autoantibodies in the vitreous of spontaneous cases of equine recurrent uveitis (ERU) has been demonstrated, the potential role of IgM reactivities during ERU pathogenesis remains unexplored. The purpose of this study was to examine the presence of IgM autoantibodies in vitreous specimens of ERU-affected horses and to test their binding specificity to intraocularly expressed proteins. Methods: To test IgM autoantibody responses to retinal tissue, vitreous samples of eye-healthy controls and ERU patients were analyzed via two-dimensional Western blot analysis with e...
Grosche A, Freeman DE, Morton AJ, Polyak MM, Matyjaszek SA.To assess the effects of ischemia and reperfusion on indicators of oxidative stress, activation of eosinophils, and apoptosis in the large colonic mucosa of horses. Methods: 40 horses. Methods: In 1 or two 20-cm-long segments of the pelvic flexure, ischemia was induced for 1 or 2 hours followed by no reperfusion or 30 minutes and 18 hours of reperfusion in anesthetized horses. Mucosal specimens were collected before (controls; n = 20 horses) and after each period of ischemia, and full-thickness tissue samples were collected after each period of reperfusion. Sections of colonic tissues were sta...
Stoltz MA, van der Merwe CF, Coetzee J, Huismans H.The subcellular localization of the minor nonstructural protein NS3 of African horsesickness virus (AHSV) has been investigated by means of immunogold electron-microscopical analysis. NS3 was observed in perturbed regions of the plasma membrane of AHSV-infected VERO cells, and its presence appears to be associated with events of viral release. These events are budding, whereby released viruses acquire fragments from the host-cell membrane, as well as by the extrusion of nonenveloped particles through the cell membrane. The membrane association of NS3 was confirmed by its detection in the disru...
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...
Bonenberger TE, Ihrke PJ, Naydan DK, Affolter VK.Immunostaining with polyclonal anti-Mycobacterium bovis (BCG) was evaluated as a single screening method for the histological identification of micro-organisms in skin biopsy specimens from various veterinary species. Confirmed archival cases infected with Mycobacteria, Nocardia, Actinobacillus, Actinomyces, Streptococcus/Staphylococcus, Dermatophilus, spirochetes, Blastomyces, Coccidioides, Cryptococcus, Histoplasma, dermatophytes, Malassezia, Sporothrix, Leishmania, Pythium, phaeohyphomycetes and Prototheca organisms were selected. A total of 70 skin biopsy specimens from the dog, cat, horse...
Bilzer T, Grabner A, Stitz L.Tissues from nine horses and one donkey suffering from natural Borna disease were investigated. Clinically, all animals demonstrated progressive reduced mentation and aggravating gait disturbances. During the clinical course anorexia and progressive loss of proprioception were observed. Cranial nerve failure was accompanied by signs of pharyngeal paralysis, sialorrhea, bruxism, and by blindness. Virologically, infectious virus was detected in the brain of all animals investigated but was not found regularly in all areas of the brain. However, in all cases, infectivity was found in the thalamus...
Fedorka CE, Loux SL, Scoggin KE, Adams AA, Troedsson MHT, Ball BA.The tolerance of pregnancy by the maternal immune system is balanced between recognition and protection. In the human this is controlled by balancing helper T cell populations (Th1, Th2) in addition to immune suppression from the regulatory arm (Tregs), but this has not been evaluated in the horse. RNA sequencing was performed on chorioallantois and endometrium of mares at 120, 180, 300 and 330 days of gestation (n = 4/stage), as well as 45-day chorioallantois (n = 4) and diestrus endometrium (n = 3). Transcripts were selected for relativity to Th1, Th2, or Treg-associated. qPCR an...
Cheng FP, Gadella BM, Voorhout WF, Fazeli A, Bevers MM, Colenbrander B.The aim of the present study was to investigate whether the induction of stallion sperm acrosome reaction (AR) by progesterone is mediated by binding of progesterone to a receptor on the sperm plasma membrane or to an intracellular progesterone receptor. Progesterone-BSA conjugate labeled with fluorescein isothiocyanate (P-BSA-FITC) in combination with a vital stain, ethidium homodimer, was applied to visualize the presence of the progesterone receptor on living spermatozoa. Alternatively, an indirect immunofluorescence technique employing a monoclonal antibody (C-262) against human intracellu...
Bae SE, Watson ED.It has been reported that oxytocin is produced not only in the hypothalamus and posterior pituitary but also in outside the classical hypothalamo-neurohypophyseal axis such as the ovary, testis, placenta and in some nonreproductive sites. In the mare, oxytocin-mRNA has been identified in the endometrium, and oxytocin and its neurophysin have been identified in the uterus. In the present study, oxytocin was localised in the endometrium of the mare at the light microscopic and ultrastructural level by immunostaining and immunogold labelling of endometrial biopsy specimens collected during estrus...
Kimble KM, Gomez G, Szule JA, Dubey JP, Buchanan B, Porter BF.An adult American Quarter Horse gelding with a history of weight loss presented with an acute onset of colic, fever, soft faeces and elevated liver enzymes. At necropsy, there were gastric mucosal masses and evidence of caecal necrosis. Histologically, the masses were lymph nodes with granulomatous inflammation and areas of liquefactive necrosis. Within and surrounding necrotic areas were free and intrahistiocytic clusters of protozoal tachyzoites. Similar but milder inflammation was evident in the spleen, lungs and liver. Necrotizing typhlitis was also evident. Immunolabelling for Toxoplasma ...
Bone reportsAugust 18, 2017
Volume 7 98-107 doi: 10.1016/j.bonr.2017.08.002
Ayodele BA, Mirams M, Pagel CN, Mackie EJ.Chondrocyte hypertrophy makes important contributions to bone development and growth. We have investigated a number of novel cartilage genes identified in a recent transcriptomic study to determine whether they are differentially expressed between different zones of equine foetal growth cartilage. Twelve genes (, , , , , , , , , , and ) were found to be more highly expressed in the zone of hypertrophic chondrocytes than in the reserve or proliferative zones, whereas and were expressed at lower levels in the hypertrophic zone than in the reserve zone. , which encodes vacuolar H ATPase (V-ATP...
Rapacz-Leonard A, Leonard M, Chmielewska-Krzesińska M, Siemieniuch M, Janowski TE.Despite their importance in mammalian reproduction, substances in the oxytocin-prostaglandins pathways have not been investigated in the horse placenta during most of pregnancy and parturition. Therefore, we quantified placental content of oxytocin (OXT), oxytocin receptor (OXTR), and prostaglandin E2 and F2 alpha during days 90-240 of pregnancy (PREG), physiological parturition (PHYS), and parturition with fetal membrane retention (FMR) in heavy draft horses (PREG = 13, PHYS = 11, FMR = 10). We also quantified OXTR and prostaglandin endoperoxide synthase-2 (PTGS2) mRNA expression ...
Pimenta J, Pires I, Prada J, Cotovio M.Melanocytic tumors are an important neoplastic disease in human and veterinary medicine, presenting large differences regarding tumor behavior between species. In horses, these tumors present a prolonged benign behavior, with rare invasiveness and metastases. In humans and small animals, invasion and metastasis have been associated with an Epithelial-Mesenchymal Transition, where the loss of E-cadherin expression plays a key role in tumor progression. This process and the role of E-cadherin have not yet been evaluated in equine melanocytic tumors. This study aimed to assess the immunolabeling ...
Benvegnen J, De Breuyn B, Gerber V, Rottenberg S, Koch C.The aim of the study was to assess the expression of the immune checkpoint inhibitor programmed death-ligand 1 (PD-L1) in equine sarcoids (ES). Programmed death-ligand 1 is expressed by various cancer cells to block T cell-mediated elimination of tumor cells. Antibodies targeting human PD-L1 were tested by immunohistochemistry for their cross-reactivity with equine PD-L1 using formalin-fixed, paraffin-embedded tissues. Our results do not support an important role of PD-L1-mediated immune evasion in ES disease and hence do not offer a rationale for anti-PD-1/PD-L1-based immunotherapy against ES...
Nelissen S, Miller AD.Granulosa cell tumors (GCTs) are common ovarian neoplasms in the mare and bitch that can be challenging to diagnose on histopathology. Inhibin has long been the standard immunohistochemical (IHC) marker for GCTs; however, anti-Müllerian hormone (AMH) has not been evaluated widely as an IHC marker in the bitch and mare. We compared the efficacy of AMH and inhibin as IHC markers in canine and equine GCTs. We selected retrospectively 18 equine and 15 canine cases. All equine tumors were dominated by a cystic pattern; canine tumors often had solid patterns. Both inhibin and AMH had similar puncta...
Hartzel DK, Arnoczky SP, Kilfoyle SJ, Stick JA.To demonstrate myofibroblasts in the accessory ligament of the deep digital flexor tendon (ie, distal check ligament) and deep digital flexor tendon of clinically normal foals. Methods: Tissue specimens from 25 foals that were necropsied for reasons unrelated to this study and unrelated to musculoskeletal disease. Methods: The distal check ligament and deep digital flexor tendon of both forelimbs were examined histologically. Myofibroblasts were identified by immunohistochemical staining specific for alpha-smooth muscle actin (alpha-SMA). Results: Most of the cells in the distal check ligament...
de Bruijn CM, Veenman JN, Rutten VP, Teske E, van Nieuwstadt RA, van den Ingh TS.This study documents the clinical, histopathological, immunohistochemical and flow-cytometric findings in five horses with cutaneous non-epidermotropic malignant lymphoma (ML). The median survival time after discovery of the first subcutaneous nodules was 3.8 years (range 2-5 years: n=4). Histologically, the cutaneous ML had a pleiomorphic structure and contained a mixture of large reticulo-endothelial cells, medium-large sized lymphoid cells with a rounded nucleus and small nucleoli, many medium sized lymphoid cells with irregular nuclei, and some small lymphoid cells. Immunohistochemically (...
The Journal of parasitologyJanuary 23, 2003
Volume 88, Issue 6 1164-1170 doi: 10.1645/0022-3395(2002)088[1164:EIOEPM]2.0.CO;2
Sofaly CD, Reed SM, Gordon JC, Dubey JP, Ogleebee MJ, Njoku CJ, Grover DL, Saville WJ.The effect of inoculation dose of Sarcocystis neurona sporocysts on the development of clinical neurologic disease in horses was investigated. Twenty-four seronegative weanling horses were subjected to the natural stress of transport and then randomly assigned to 6 treatment groups of 4 horses each. Horses were then immediately inoculated with either 10(2), 10(3), 10(4), 10(5), or 10(6) S. neurona sporocysts or placebo using nasogastric tube and housed indoors. Weekly neurologic examinations were performed by a blinded observer. Blood was collected weekly for antibody determination by Western ...
Carrasco L, Mendez A, Jensen HE.An atypical case of chronic equine bronchopulmonary aspergillosis with an unusual hyphal morphology was diagnosed in a horse with Cushing's syndrome. Because of the hyphal localization in chronic ectatic bronchi and bronchioles, and juxtabronchiolar processes, the observed type of aspergillosis is similar to 'saprophytic bronchopulmonary aspergillosis' or 'semi-invasive pulmonary aspergillosis' in humans. The aetiological diagnosis of aspergillosis was accomplished by the application of a panel of monospecific polyclonal and monoclonal antibodies in immunohistochemical techniques.
Yasui T, Fukui K, Nara T, Habata I, Meyer W, Tsukise A.The present study revealed in detail the subcellular localization of lysozyme and beta-defensin in the apocrine glands of the equine scrotal skin, a specific body region. The apocrine glandular cells were equipped with a varying number of secretory granules, a well-developed Golgi apparatus and abundant cisternae of the rough endoplasmic reticulum within their cytoplasm. In these cells, reactive gold particles representing lysozyme were detectable in the secretory granules as well as the Golgi apparatus and elements of the rough endoplasmic reticulum. Additionally, the antimicrobial peptide gr...
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.
Klein C, Scoggin KE, Troedsson MH.Establishment of pregnancy is critically dependent upon a precisely orchestrated embryo-maternal interaction leading to a receptive uterine environment. The up-regulation of the interferon-stimulated protein 15 kDa (ISG15) during pregnancy has been described in various species and has been hypothesized to be part of the molecular repertoire that makes the uterus receptive to conceptus development. In the current study, the expression of ISG15 and enzymes involved in ISG15ylation was examined at the mRNA and protein level in equine endometrium at Day 14 of the luteal phase and at Day 14 and 50 ...
Gomes RG, Silva CB, González SM, Oliveira RL, Max MC, Lisboa LA, Barreiros TRR, Santos MM, Sarapião FD, Gastal EL, Seneda MM.The effect of different concentrations of alpha lipoic acid (ALA) on the development and morphology of preantral follicles, as well as the proliferative activity of granulosa cells, was assessed after short-term culture. Ovaries (n = 5) of five seasonal anestrous mares were harvested in a local abattoir. At the laboratory, nine ovarian fragments (5 × 5 × 1 mm) from each animal were used. One fragment was immediately fixed in Bouin and subjected to histological and immunohistochemistry (proliferating cell nuclear antigen, PCNA) analyses (noncultured group; D0 = day 0). The other eigh...
Gumber S, Baia P, Wakamatsu N.A 15-year-old female gray Appaloosa horse was presented with history of a mass over the right dorsal commissure of vulva for the past 7 months. Based on histopathological examination, and positive staining with factor VIII-related antigen, vimentin, and Verhoeff-van Gieson stain, the vulvar mass was diagnosed as hemangiosarcoma with marked solar elastosis.
Garcia-Suarez O, Germanà G, Naves FJ, Ciriaco E, Represa J, Vega JA.The medial wall of the vomeronasal organ (VNO) is lined with a sensory epithelium that is closely related to the olfactory epithelium, which is developed from the olfactory placode. It undergoes continuous replacement during its life span. In other sensory epithelia, cell proliferation is under the control of some trophic factors. Whether these proteins are involved in the continuous turnover of the VNO epithelium is unknown. This study approaches this topic by analyzing the occurrence of signal-transducing receptor proteins for neurotrophins (Trk proteins) and epidermal growth factor (EGFr). ...
Ekman S, Lindahl A, Rüetschi U, Jansson A, Björkman K, Abrahamsson-Aurell K, Björnsdóttir S, Löfgren M, Hultén LM, Skiöldebrand E.Molecular serum markers that can identify early reversible osteoarthritis (OA) in horses are lacking. Objective: We studied serum concentrations of a novel cartilage oligomeric matrix protein (COMP) neo-epitope in horses subjected to short-term exercise and with acute lameness. The effects of circadian rhythm and age were also evaluated. Methods: Longitudinal studies in healthy horses and cross-sectional comparison of lame and non-lame horses. Methods: Sera were collected from five horses before and after short-term interval exercise and during full-day box rest. Sera from 32 acutely lame hors...
Patrick DJ, Kiupel M, Gerber V, Carr EA.A 2-year-old female Miniature Horse that presented with a history of progressive weight loss, depression, and diarrhea was diagnosed at necropsy with a highly malignant abdominal neoplasm involving the left ovary, kidneys, adrenal glands, intestines, and various abdominal and thoracic lymph nodes. Microscopic examination of these masses revealed large pleomorphic cells that stained positive for vimentin and inhibin and negative for epithelial membrane antigen and placental alkaline phosphatase. Ultrastructural examination of the cells revealed a high nucleocytoplasmic ratio and indented euchro...
Masri MD, Alda JL, Dubey JP.Sarcocystis neurona-like schizonts were found in sections of brain and spinal cord of two ataxic horses from Brazil. The diagnosis was supported by staining with anti-Sarcocystis serum in an immunohistochemical test. One of the affected horses was born in Argentina and raised in Brazil, and the other horse was born and raised in Brazil.
Wagner B.Host immune analyses require specific reagents to identify cellular and soluble components of the immune system. These immune reagents are often species-specific. For horses, various immunological tools have been developed and tested by different initiatives during the past decades. This article summarizes the development of well characterized monoclonal antibodies (mAbs) for equine immune cells, immunoglobulin isotypes, cytokines, and chemokines.
