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.
Slovis NM, Watson JL, Affolter VK, Stannard AA.Three horses were presented with a history of having developed raised cutaneous nodules, within 24-48 hours, in areas of previous injections using standard silicone-coated hypodermic needles. Skin biopsies were taken from a selected cutaneous nodule from all horses for histopathologic evaluation. Histologically, the nodules were consistent with a diagnosis of equine eosinophilic granuloma. A hypersensitivity reaction to the silicone, or another component of the coating formulation, was hypothesized to be responsible for these lesions. Two horses were experimentally injected using both coated a...
Pérez J, Mozos E, Martín MP, Day MJ.The distribution of T (CD3), B (CD79) lymphocytes, immunoglobulin (IgG, IgM and IgA)-producing plasma cells, macrophages (lysozyme, Mac387) and MHC Class II antigen was analysed in the inflammatory infiltrate associated with 19 equine squamous cell carcinomas (SCCs) and six cases of precancerous lesions (actinic keratosis). The SCCs came from the penis (11 cases), conjunctiva (four), skin (two), nasal cavity (one) and oral cavity (one). Seven cases were well-differentiated and 12 moderately differentiated. Nine cases showed no invasion of peritumoral deep tissues (locally invasive), whereas th...
Walter I, Nowotny N.Effects of infection with two different strains of equine herpes virus type 1 (EHV-1; Piber 178/83, Kentucky D) on the cytoskeleton of Vero cells were investigated immunohistochemically, and evaluated by confocal laser scanning microscopy. Twenty four hours post EHV-1 infection the assembly of the microtubulus system of Vero cells was heavily disturbed. The Golgi region was dispersed into vesicles spread throughout the cytoplasm as demonstrated by WGA lectin binding. Other cytoskeletal elements such as cytokeratin, vimentin, and filamentous actin (F-actin) were not affected by EHV-1 infection....
Gilger BC, Malok E, Cutter KV, Stewart T, Horohov DW, Allen JB.Equine recurrent uveitis (ERU), a chronic, recurrent inflammation primarily of the anterior uveal tract, is the most common cause of blindness in horses. Recently, T-lymphocytes have been found to be the most numerous cell type to infiltrate the anterior uveal of horses with ERU. In the present study, we characterized the T-lymphocyte population in the anterior uveal tract of eyes of horses with chronic ERU by evaluating the microscopic appearance (histopathologic features), the T-lymphocyte subsets, and the relative levels and amounts of T-lymphocyte cytokine mRNA in the anterior uvea. Seven ...
Gerstenberg C, Allen WR, Stewart F.Immunohistochemical detection of the proliferation marker Ki-67 antigen was used to monitor mitotic activity in the endometrium of mares. The monoclonal antibody MIB1 was validated for use on equine tissues by demonstrating its reaction with activated peripheral blood lymphocytes, and endometrial biopsies were recovered from 26 non-pregnant mares at selected stages during the reproductive cycle. The proportion of positively stained nuclei was counted in five random areas on each histological section to determine the percentage and type of proliferating cells. Multiplication rates in the types ...
Oaks JL, Ulibarri C, Crawford TB.Equine infectious anaemia virus (EIAV) infection of horses is characterized clinically by recurrent episodes of fever, thrombocytopenia and anaemia. In vivo, the only site of virus replication that has been previously demonstrated for EIAV is the tissue macrophage. In this study, in situ hybridization for EIAV was combined with immunohistochemistry for cell-type-specific markers to identify infected endothelial cells. EIAV-infected endothelial cells and macrophages were detected in horses infected with either virulent wild-type or with weakly virulent tissue culture-adapted strains of EIAV. Th...
Shikichi M, Kitamura HP, Yanase H, Konno A, Takahashi-Iwanaga H, Iwanaga T.The synovial membrane displays a superficial cellular lining composed of two types of synoviocytes: "absorptive" macrophages (type A cells) and "secretory" fibroblast-like cells (type B cells). The types are intermingled and extend a variety of processes, rendering the cellular architecture of the synovial membrane difficult to visualize. Previous electron microscopic and histochemical studies failed to demonstrate the entire shape of synoviocytes, except our immunohistochemical study for protein gene product 9.5 in the horse joint. The present SEM study is the first to demonstrate the three-d...
Ripoll CM, Remondegui CE, Ordonez G, Arazamendi R, Fusaro H, Hyman MJ, Paddock CD, Zaki SR, Olson JG, Santos-Buch CA.Between November 1993 and March 1994, a cluster 6 pediatric patients with acute febrile illnesses associated with rashes was identified in Jujuy Province, Argentina. Immunohistochemical staining of tissues confirmed spotted fever group rickettsial infection in a patient with fatal disease, and testing of serum of a patient convalescing from the illness by using an indirect immunofluorescence assay (IFA) demonstrated antibodies reactive with spotted fever group rickettsiae. A serosurvey was conducted among 16 households in proximity to the index case. Of 105 healthy subjects evaluated by IFA, 1...
Tunev SS, Ehrhart EJ, Jensen HE, Foreman JH, Richter RA, Messick JB.An 18-year-old Morgan mare was presented to the Veterinary Medical Teaching Hospital, University of Illinois, with a 10-day history of watery diarrhea, depression, and dysphagia. On admission, the animal was severely dehydrated, depressed, and unable to swallow and had no clinical signs of diarrhea. The respiratory and heart rate and body temperature were within normal limits. Following fluid therapy, the mare developed severe watery diarrhea and continued to be depressed, incoordinated, and dysphagic. The animal died on the fourth day after admission and was sent to the Laboratories of Veteri...
Rivero JL, Serrano AL, Barrey E, Valette JP, Jouglin M.Combined methodologies of enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE), immunoblotting, traditional myofibrillar ATPase (mATPase) histochemistry and immunocytochemistry of whole biopsied samples were used to study myosin heavy chain (MHC) isoforms in the equine gluteus medius muscle. The ELISA technique allowed the quantification of the three MHC isoforms known to be present in different horse muscles: slow (MHC-I) and two fast (termed MHC-IIA and MCH-IIX). The SDS-PAGE method resolved MHCs in three bands: MHC-I, MHC-IIX and M...
Hong CB, Van Meter PW, Latimer CL.A malignant rhabdoid tumour was diagnosed in the orbit of a 2-year-old Thoroughbred filly. The neoplasm, which was very aggressive, was present in nearly every part of the ocular and periocular structures and had spread to the lymph nodes of the head and neck, the salivary glands and the subcutaneous tissues around the eye. The neoplasm was composed of polygonal cells with abundant eosinophilic cytoplasm. Many cells had a large, vesiculate, indented nucleus and contained a paranuclear globular inclusion. Ultrastructurally, the inclusions were seen to consist of whorls of intermediate filaments...
Linnane L, Serrano AL, Rivero JL.The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in muscle biopsies from the gluteus medius of adult untrained horses by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies and standard myofibrillar ATPase (mATPase) histochemistry. Percutaneous needle biopsies were taken at 3 depths (20, 40 and 60 mm) from 4 4-y-old Andalusian stallions. The percentage of 'pure' I MHC fibres increased whereas that for pure IIX MHC fibres decreased from the most superficial to the deepest sampling sit...
Kalsow CM, Dubielzig RR, Dwyer AE.Pinealitis accompanying uveitis is well established in laboratory models of experimental autoimmune uveoretinitis. In naturally occurring uveitis, pinealitis has been demonstrated in the pineal gland from a mare with active uveitis and is suspected in some human uveitides. We have evaluated pineal glands from horses with various stages of uveitis for signs of immunopathology accompanying spontaneous uveitis. Methods: Pineal glands from 10 horses with uveitis and from 13 horses without uveitis were evaluated for histochemical (H&E, collagen) and immunohistochemical (MHC class II antigen exp...
Rungsipipat A, Tateyama S, Yamaguchi R, Uchida K, Miyoshi N.An immunohistochemical study of various visceral organs of normal adult dogs, cats, pigs, horses, cows, and chickens (five of each species) and of 185 spontaneous canine tumours was carried out using paraffin wax sections and a commercially available antibody to the human c- yes oncogene product. Among the adult normal tissues of six animal species, epithelial cells of the proximal and distal renal tubules, the myocardium, hepatocytes, cerebellar Purkinje cells and adrenal cortical cells were positive for c- yes product. Among the foetal tissues of dogs and chickens, a positive reaction was ob...
Hudson NP, Pearson GT, Kitamura N, Mayhew IG.The interstitial cells of Cajal (ICC) are c-kit immunoreactive cells of the gastrointestinal tract which are suggested to have a role in the control of intestinal motility. Cells with c-kit immunoreactivity have not been previously described in the gastrointestinal tract of the horse. Immunoreactivity for c-kit was revealed using immunohistochemical labelling with an anti-c-kit polyclonal antibody. Sections of normal gastrointestinal tissue were examined from 13 anatomically defined sites from stomach to small colon taken from horses free from gastrointestinal disease. Three types of c-kit imm...
Watson ED, Buckingham J, Björkstén TS.Oxytocin has been identified in the ovaries of many species
including ruminants (Wathes and Swann 1982; Rodgers et al.
1983), pigs (Jarry etal. 1990), sub-human primates (Khan-Dawood
1998), rats (Ho and Lee 1992) and women (Schaeffer et al. 1984).
In these species ovarian oxytocin has several roles, including
control of luteal regression (Fht and Sheldrick 1983) and possibly
luteinisation (Aladin Chandrase.kher and Fortune 1990; Eispanier et
al. 1997), and involvement in ovulation (viggiano et al. 1989).
In the mare, circulating oxytocin is reported to be high in
oestrus and early dio...
Blunden AS, Gower SM.Lungs were obtained from 16 Thoroughbred horses, aged 1 day to 2 years, which had died or been humanely killed for reasons unrelated to disease of the lower respiratory tract. The lungs were then subjected to a histological and immunohistochemical examination of the humoral immune system. At birth there was no evidence of organized lymphoid tissue, and lymphocytes and plasma cells were virtually absent in all tissue compartments in the first week of life. However, by 12 weeks, foals exhibited well developed bronchus- and bronchiole-associated lymphoid tissue, but this had regressed progressive...
Banks EM, Kyriakidou M, Little S, Hamblin AS.Leucocytes in the lung epithelium play an important role in the ability of an animal to respond appropriately to inhaled pathogens. The distribution of lymphoid and myeloid cells associated with the lung epithelium was examined immunohistochemically throughout the respiratory tract of four horses, comprising two adults from an abattoir, one pregnant mare, and her fetus (in the final stage of gestation). Cross and tangential cryosections were labelled with monoclonal antibodies against T-cell, B-cell, macrophage/dendritic myeloid cell, and major histocompatibility Class (MHC) II surface antigen...
Kitamura HP, Yanase H, Kitamura H, Iwanaga T.Fibroblast-like (Type B) synoviocytes are cells in the synovial membrane that are responsible for production of both synovial fluid and the extracellular matrix in the synovial intima. Immunostaining of the horse synovial membrane for protein gene product (PGP) 9.5, which is a neuron-specific ubiquitin C-terminal hydrolase, demonstrated selective localization of the immunoreactivity in a synoviocyte population different from acid phosphatase-positive Type A synoviocytes. The immunoreactive cells were lined up in the synovial intima and extended dendritic processes towards the joint cavity to f...
Jones CJ, Wooding FB, Dantzer V, Leiser R, Stoddart RW.The glycosylation of the equine interhaemal barrier and areola was studied throughout the period of gestation. Placentae of 35, 37, 50, 119, 152, 200, 280 and 300 days gestation were investigated, using semithin plastic embedded sections and a panel of 15 biotinylated lectins with an avidin-peroxidase revealing system. Glycosylation of the trophoblast and maternal epithelium showed the most change during the first 50 days of gestation, being associated with the initial stages of adhesion and attachment. In the trophoblast, non-bisected tri/tetraantennary complex N-glycan was only evident after...
Wattle O.The equine skin and its appendages (chestnut, hoof capsule, ergot, sebaceous glands, sweat glands and hair) consist mainly of keratinocytes. The intermediate filament cytoskeleton of these cells in involved in specialised functions, such as mechanical co-ordination of the cytoskeleton of the cell or tissue. In this study, 7 monoclonal antibodies, one polyclonal antibody and immunoblot analysis were used to characterise cytokeratins (separated by 1- and 2-dimensional gel electrophoresis) from the hoof wall and chestnut. The tissue distribution of these cytokeratins was studied by immunohistoche...
Kalsow CM, Dwyer AE.Equine uveitis is a spontaneous disorder of horses that can serve as a model for the study of human uveitis. Although the initial presentation is that of an anterior uveitis, retinal involvement has been noted in some cases. We report here the immunohistopathology of retinas from horses with uveitis. Methods: Sections of eyes recovered from horses with naturally occurring uveitis and from Shetland ponies with experimental leptospira-induced uveitis were stained by hematoxylin and eosin for histopathological evaluation. Immunohistochemistry was used to evaluate retinas for MHC Class II antigen ...
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...
Dagleish MP, Pemberton AD, McAleese SM, Thornton EM, Miller HR, Scudamore CL.Equine alpha-1-proteinase inhibitor (API) consists of three, occasionally four, serum glycoproteins. This study investigated the immunohistochemical localisation of equine API in paraformaldehyde fixed, paraffin embedded equine tissue samples of liver, lung, stomach, pancreas, jejunum and colon in five horses using affinity purified sheep polyclonal and protein A purified mouse monoclonal antibodies, whose specificities were verified by Western blotting. Exposing tissue sections to boiling citrate buffer greatly enhanced antigen recovery and improved immunostaining with both antibodies, result...
Hernández-Vidal G, Jeffcott LB, Davies ME.A polyclonal antiserum raised in sheep against human cathepsin B was tested for specificity and cross-reactivity with the horse homologue by SDS-PAGE and Western blotting, prior to being used for immunolocalization of the enzyme in equine articular cartilage. In Western blots, the antiserum recognized the 30 kDa single chain and 25 kDa heavy chain of the mature enzyme in purified bovine cathepsin B, and corresponding bands at 32 and 27 kDa in equine chondrocyte and fibroblast lysates. This antiserum was then used to compare the expression and distribution of cathepsin B in normal and dyschondr...
Kalsow CM, Albrecht TR, Steinberg NP, Lunn DP.Immunohistochemical evaluation of equine tissue necessitates the use of antibodies reactive with cells from a heterogeneous population. Lymphoid tissues from 12 horses were fixed in Bouin's fluid, ethanol or formalin and examined for immunohistochemical reactivity with anti-equine and anti-human monoclonal antibodies (MAbs) specific for MHC Class II antigens, T and B lymphocytes, and macrophages. Only a few of the anti-equine MAbs tested were reactive with fixed, paraffin wax-embedded tissue. Anti-human MAbs expanded the desired range of reactivity and increased the consistency in different an...
Hamir AN, Tornquist SJ, Gerros TC, Topper MJ, Dubey JP.Equine protozoal myeloencephalitis (EPM) was clinically diagnosed in a 20-year-old horse with severe ataxia. The cerebrospinal fluid was positive for Sarcocystis neurona antibodies by western blot. The horse was administered corticosteroids to facilitate in vitro culture of S. neurona from its spinal cord following necropsy. Microscopic lesions of EPM were present in the brain and in the spinal cord, including multifocal inflammatory cellular infiltrates and several large groups of protozoa. Immunohistochemical, and light and electron microscopic examinations revealed that the protozoa were Ne...
Nagamine N, Nambo Y, Nagata S, Nagaoka K, Tsunoda N, Taniyama H, Tanaka Y, Tohei A, Watanabe G, Taya K.To determine the source of circulating inhibin and estradiol-17beta during the estrous cycle in mares, the cellular localization of the inhibin alpha, betaA, and betaB subunits and aromatase in the ovary was determined by immunohistochemistry. Concentrations of immunoreactive (ir-) inhibin, estradiol-17beta, progesterone, LH, and FSH in peripheral blood were also measured during the estrous cycle in mares. Immunohistochemically, inhibin alpha subunits were localized in the granulosa cells of small and large follicles and in the theca interna cells of large follicles, whereas inhibin betaA and ...
Romeike A, Brügmann M, Drommer W.Despite extensive clinical research, the etiology of equine recurrent uveitis (ERU) is still unknown. After an immunologic pathogenesis was established in recurrent uveitis in humans, a similar pathogenic mechanism was assumed to exist in ERU. To investigate whether immunopathologic mechanisms are involved in ERU, 20 eyes of 15 horses with ERU were examined immunohistochemically with a T cell marker, B cell marker, and anti-major histocompatibility complex (MHC) class II antibodies. Twenty-six eyes of 20 horses were used for investigation of MHC class II antigen expression in normal equine eye...
Hoshiba H, Sinowatz F.Spermadhesins are proteins with various functions in sperm capacitation and zona pellucida binding. In this study the cellular localization of the spermadhesin AWN-1 has been examined in the equine male genital tract. Results obtained by immunohistochemical methods reveal that in the horse AWN-1 is synthesized in spermatogonia, in the rete testis, the ductus epididymidis and the seminal vesicles. These findings indicate that the cellular origin of spermadhesins is species-specific.
Lorenzo PL, Liu IK, Illera JC, Picazo RA, Carneiro GF, Illera MJ, Conley AJ, Enders AC, Illera M.Epidermal growth factor (EGF) has been reported to promote different functions in mammalian ovaries, including oocyte maturation. The aim of the present study was to establish: that EGF influences oocyte maturation in ovine and equine, that a tyrosine kinase-dependent intracellular mechanism mediates EGF effect and, that EGF-R receptor is detectable in ovarian follicles by immunohistochemistry methods. Selected ovine and equine oocytes were aspirated from 2-5 mm (ovine) or 25 mm (equine) follicles and cultured in TCM 199 for 22 (ovine) or 36 hours (equine). They are then subjected to culture w...
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...
Nagel H, Lang H, Sole Guitart A, Lean N, Allavena R, Sprohnle-Barrera C, Young A.This case series presents a 20-year-old Quarter horse gelding and a 10-year-old Thoroughbred gelding with a histologic diagnosis of aggressive submural neoplasia, including an anaplastic sarcoma and a squamous cell carcinoma respectively. The current case series describes these neoplasias with radiography, computed tomography, magnetic resonance imaging and histopathology with immunohistochemistry. The purpose of this case series is to identify the clinical pattern of chronic reoccurring lameness in these cases and highlight the appearance of expansile osteolytic pathology of the distal phalan...
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 ...
Veado HC, Silva AS, Fagundes JLA, Bittencourt AA, de Castro MB, Câmara ACL.Skin lesions are among the leading diseases in equids, and the urogenital region is considered the third most frequently affected body location with tumors and tumor-like skin lesions in horses. Tumor-like skin lesions or multiple tumors of different types in the equine prepuce and penis are a challenging clinical presentation leading to misdiagnosis and subsequent ineffective treatment. An 18-year-old 395-kg male mixed-breed gelding was referred for evaluation of preputial enlargement. Clinical examination revealed a 2.5 × 2.0 × 0.8 cm proliferative ulcerated lesion on the penile mucosa...
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...
Griffiths IR, Lusk SA, Kyriakides E, Smith S.Neurofilaments (NF) are composed of three polypeptides of differing molecular size, termed NF-L, NF-M and NF-H. The NF-H and, to a lesser degree, NF-M components are phosphorylated. In the majority of normal neurones, the location of phosphorylated NF is confined to neuronal processes, particularly the axon, and excluded from the perikaryon. Cell bodies of autonomic neurones of the rat do not contain phosphorylated NF. In many disease states, phosphorylated NF accumulate in the neuronal cell body and therefore in most circumstances their presence indicates abnormality. This paper reports that ...
Scott P, Witonsky S, Robertson J, Daft B.Equine protozoal myeloencephalitis (EPM), caused by Sarcocystis neurona infection in the central nervous system (CNS), affects up to 1% of all horses during their lifetimes. Neither the protective immune response nor the immunopathology associated with the disease is well understood. To begin to clarify the pathogenesis of the disease, immunohistochemical staining for B and T lymphocytes was performed on spinal cord sections obtained from 17 horses, all of which were all positive for S. neurona based on immunohistochemical staining. Fifteen of the 17 horses included in the study were killed du...
Li JR, Wang W, Shi FX.The effects of equine chorionic gonadotropin (eCG) on follicular development and ovulation in cyclic guinea pigs were investigated by histological and immunohistochemical analyses. Three groups of guinea pigs (n=12) were administrated subcutaneously with saline, 20 or 50 IU of eCG, respectively, on cyclic Day 12 (Day 1=vaginal openings). Ovaries were collected at 4 and 8 d after administration (6 animals per group each time). The eCG administration induced significant and distinct morphological changes in the ovaries, as it promoted the luteinization of granulosa cells, but not follicular deve...
Sellers RS, Toribio RE, Blomme EA.Idiopathic systemic granulomatous disease, which has been reported in horses, cattle and human beings, is characterized by perivascular granulomatous and lymphoplasmacytic inflammation in many organ systems. Diagnosis is based on the exclusion of possible viral, fungal or bacterial causes. The disease was identified in a miniature pony with widespread lymphoplasmacytic and granulomatous inflammation, special staining techniques having revealed no evidence of any aetiological agent. Skin lesions, which were severe, consisted of hyperkeratosis and serocellular crust formation, with inflammatory ...
Webb TL, Burnett RC, Avery AC, Olver CS.Characterization of anemia in horses presents a challenge, as they do not release reticulocytes into peripheral blood. Transferrin receptor (TfR) expression is highest on erythroid cells in people and rats, and measurement of a soluble serum form (sTfR) is used to quantify erythropoiesis in these species. We hypothesized that equine TfR (eTfR) expression is similar in quantity and distribution to that in these other species and thus has potential for characterization of the regenerative response in anemic horses. Objective: This study was conducted to clone and sequence the eTfR gene and measu...
Fortin-Trahan R, Lemirre T, Santschi EM, Janes JG, Richard H, Fogarty U, Beauchamp G, Girard CA, Laverty S.There is a knowledge gap about how equine MFC subchondral radiolucencies (SR) arise and evolve. Osteoclasts are believed to have a role but have not been studied in situ. Objective: To measure and compare osteoclast density and the percentage of chondroclasts in healthy and MFC SR specimens from juvenile Thoroughbreds. Methods: Cadaveric study. Methods: Medial femoral condyles (MFC) from a tissue bank of equine stifles were studied. Inclusion criteria were MFCs (≤8 months old) with a computed tomography SR lesion and histological focal failure of endochondral ossification (L group). Contral...
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...
Sayasith K, Bouchard N, Boerboom D, Brown KA, Doré M, Sirois J.Ovulation is accompanied by a marked infiltration of leukocytes into thecal layers after the gonadotropin surge. P-selectin is known to play a critical role in the initial steps of leukocyte recruitment from the bloodstream during inflammation. Thus, the objective was to investigate the potential regulation of P-selectin by gonadotropins in equine preovulatory follicles. The full-length equine P-selectin cDNA was cloned by a combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. Results showed that equine P-selectin cDNA encodes...
Martínez-Galisteo A, Diz A, Agüera E, Vivo J.34 Andalusian foals of both sexes were divided into three age-groups (A = mean age 1 month, B = 7 months, C = 14 months). Samples of the right m. gluteus medius were stained for m-ATPase and NADH-TR in order to determine fibre type composition and size as well as the relative area occupied by each type. Results revealed no statistically significant variation in the proportion of type I fibre among the three age-groups. Significant differences were recorded, however, for type II fibres; an increase in the proportion of IIA fibres was accompanied by a decrease in IIB ones, the difference being m...
Hyyppä S, Karvonen U, Räsänen LA, Persson SG, Pösö AR.To study the effects of nandrolone laureate (19-nortestosterone) on muscle hypertrophy and concentration of androgen receptors (AR), biopsy specimens were taken from the middle gluteal muscle of 6 Finnhorse trotters (geldings and mares) undergoing training before, immediately after, and 13 weeks after a 14-week treatment with nandrolone. Another 6 similarly trained horses served as controls. An additional 10 mares and 10 geldings were used to study annual variation in muscle concentration of AR. AR was immunohistochemically localized in the nuclei. AR concentration remained constant during the...
Banse HE, Piero FD, Andrews FM, Garcia-Abarca N, Watanabe TTN.To compare small intestinal inflammation with gastric inflammation in horses with and without equine gastric glandular disease (EGGD), we evaluated endoscopic, macroscopic, and microscopic findings of the glandular stomach and microscopic findings of the small intestine. Methods: 36 horses. Methods: Horses underwent endoscopy and were scored for EGGD. After euthanasia, stomachs were collected and macroscopically evaluated. Normal pyloric mucosa, glandular lesions, and small intestinal (duodenum, mid-jejunum, and ileum) samples were collected and processed for microscopic examination. Cellular ...
De Castilho T, Rosa GDS, Stievani FC, Apolônio EVP, Pfeifer JPH, Altheman VG, Palialogo V, Santos NJD, Fonseca-Alves CE, Alves ALG.Tendinopathies account for a substantial proportion of musculoskeletal injuries. To improve treatment outcomes for partial and total tendon ruptures, new therapies are under investigation. These include the application of mesenchymal stem cells (MSCs) and biocompatible scaffolds derived from the Extracellular Matrix (ECM). Synthetic polymer hydrogels have not demonstrated results as promising as those achieved with ECM hydrogels sourced from the original tissue. This study aimed to evaluate the biocompatibility of a hydrogel formulated from equine tendon ECM. Six horses were administered three...
Leemans B, Gadella BM, Marchand JHEAM, Van Soom A, Stout TAE.We recently developed re-differentiated equine oviduct epithelial cell (REOEC) monolayers demonstrating various in vivo morphological characteristics, but lacking secondary ciliation. In this study, we evaluated the effects of fetal bovine serum, reproductive steroid hormones, Wnt- and Notch ligands and inhibitors, and different EOEC seeding densities, in both conventional wells and on microporous membranes, on EOEC morphology and, in particular, secondary ciliation. REOEC monolayers were assessed by confocal microscopy after combined staining of nuclei, cilia and the cytoskeleton. Only Wnt li...
Brodesser DM, Kummer S, Eichberger JA, Schlangen K, Corteggio A, Borzacchiello G, Bertram CA, Brandt S, Pratscher B.The ability of human melanoma cells to switch from an epithelial to a mesenchymal phenotype contributes to the metastatic potential of disease. Metalloproteinases (MPs) are crucially involved in this process by promoting the detachment of tumor cells from the primary lesion and their migration to the vasculature. In gray horse melanoma, epithelial-mesenchymal transition (EMT) is poorly understood, prompting us to address MP expression in lesions versus intact skin by transcriptome analyses and the immunofluorescence staining (IF) of gray horse tumor tissue and primary melanoma cells. RNAseq re...
Verhaar N, Reineking W, Hewicker-Trautwein M, Grages AM, Kästner SBR, Geburek F.To evaluate the use of laser Doppler flowmetry and spectrophotometry (LDFS) for large intestinal viability assessment in horses with naturally occurring large intestinal strangulations. Methods: By use of LDFS, intestinal microperfusion was quantified as tissue oxygen saturation (tSo2), hemoglobin (tHB), and blood flow (tBF) in cases with large colon volvulus and small colon strangulations undergoing colic surgery (n = 17). Intestinal biopsies were taken from the pelvic flexure in all large colon cases and in small colon cases that underwent intraoperative euthanasia. Measurements were compare...
Martano M, Power K, Cuccaro B, Razzuoli E, Maiolino P, Restucci B.The equine sarcoid is the most common skin neoplasia of fibroblastic origin in horses, characterized by an excessive accumulation of extracellular matrix produced by sarcoid fibroblasts under hypoxic condition. Neoplastic cells can adapt to hypoxia by using alternative energy sources, particularly those that arise from fatty acid oxidation (FAO). The Carnitine Palmitoyl Transferase 1A (CPT1A) belongs to Carnitine System (CS) and promotes the entrance of fatty acids into the mitochondria for β-oxidation. In this study, CPT1A expression was comparatively addressed in 25 equine sarcoids and 5 no...
Kambayashi Y, Nemoto M, Ochi A, Kishi D, Ueno T, Tsujimura K, Bannai H, Kawanishi N, Ohta M, Suzuki T.In this study, equine intestinal enteroids (EIEs) were generated from the duodenum, jejunum, and ileum and inoculated with equine coronavirus (ECoV) to investigate their suitability as in vitro models with which to study ECoV infection. Immunohistochemistry revealed that the EIEs were composed of various cell types expressed in vivo in the intestinal epithelium. Quantitative reverse-transcription PCR (qRT-PCR) and virus titration showed that ECoV had infected and replicated in the EIEs. These results were corroborated by electron microscopy. This study suggests that EIEs can be novel in vitro ...
Stewart AS, Kopper JJ, McKinney-Aguirre C, Veerasamy B, Sahoo DK, Freund JM, Gonzalez LM.Gastrointestinal disease is a leading cause of death in mature horses. A lack of in vitro modeling has impeded the development of novel therapeutics. The objectives of this study were to develop and further characterize a small intestinal monolayer cell culture derived from equine jejunum including establishing normal measurements of intestinal permeability and restitution. Three-dimensional enteroids, derived from postmortem sampling of equine jejunum, were utilized to develop confluent epithelial monolayers. The presence of differentiated intestinal epithelial cell types and tight junctions ...
Risvanli A, Hatipoglu F, Salykov R, Timurkaan N, Kadiralieva N, Kasymalieva KK, Seker I, Ekinci E.In the current study, we aimed to determine the distribution of telocytes in the ovary, oviduct and uterus of Kyrgyz mares depending on whether they were pregnant. We also studied the progression during the first 6 months of pregnancy. To this end, ovary, oviduct and uterus tissues from 53 Kyrgyz mares were obtained post-mortem. The mare tissue samples were divided into two groups: pregnant (n = 43) and non-pregnant (n = 10). Telocyte levels in the tissues were determined using light microscopic and immunohistochemical methods. Subsequently, the data acquired were evaluated based on ...
Santiviparat S, Suthithanakom S, Bhanpattanakul S, Srisuwattanasagul S, Melde K, Stout TAE, Tharasanit T.Advances in endometrial tissue engineering have enabled the combination of modified scaffolding materials with modern cell culture technologies. Genipin and three-dimensional (3D) printing have advanced cell-tissue engineering by enabling the precise layering of cell-containing matrices while ensuring low cytotoxicity. This study aimed to advance equine endometrial tissue engineering by designing customized collagen scaffolds using 3D printing technology, while optimizing the genipin concentration to avoid toxicity. Genipin was tested at concentrations of 4, 2, 1, 0.5, 0.25, 0.125, and 0 mM on...
Medica AJ, Swegen A, Seifi-Jamadi A, McIntosh K, Gibb Z.Oxidative stress plays a critical role in regulating sperm function, yet species-specific antioxidant mechanisms remain poorly understood. This study compared hydrogen peroxide (H2O2) tolerance in horse and human sperm and investigated the roles of catalase and glutathione peroxidase (GPx) in horses. A H2O2 dose-response assay (0-2000 µM) showed that horse sperm were significantly more resistant to oxidative damage, with an IC50 for progressive motility over 14-fold higher than that of human sperm (391.6 µM vs. 27.3 µM). Horse sperm also accumulated more intracellular H2O2 without loss of m...
Zamith Cunha R, Gobbo F, Morini M, Zannoni A, Mainardi C, D'arpe L, Gramenzi A, Chiocchetti R.A growing body of evidence indicates that the endocannabinoid system (ECS) is essential for controlling the homeostasis of the skin and that the ECS is modified in the presence of skin disease. It is plausible to expect that the lamellar junction of the hoof expresses cannabinoid receptors and that their expression could be affected by lamellar disease. The goal of this study was to characterise the cannabinoid receptor type 1 (CB1R) and type 2 (CB2R) and the G protein-coupled receptor 55 (GPR55) within the dermo-epidermal junction of the hooves of healthy and laminitic horses. The expression ...
Valberg SJ, Porter A.Muscle biopsy is often required to provide a definitive diagnosis for neuromuscular disorders and can be performed using open surgical or percutaneous needle biopsy techniques. Fresh samples that are subsequently frozen in the laboratory are preferred by laboratories engaged in research, whereas formalin-fixed muscle is processed by diagnostic laboratories with specialized tinctorial and immunohistochemical techniques. Interpretation by an experienced histopathologist, combined with the signalment, history, and clinical input, is essential for establishing a diagnosis. This article outlines be...
Marchio SP, El-Sheikh Ali H, Scott MA, Barbosa Fernandes C, Scoggin KE, Troedsson M, Boakari Y.Despite its critical role in protecting the fetus, the amniotic membrane remains poorly understood in the context of disease response. The equine amniotic membrane is an important physical barrier to the amniotic compartment, and there is evidence that it may contribute to surfactant synthesis. Surfactants are essential for normal fetal lung development, and disruptions in its availability may be linked to future neonatal complications. Therefore, understanding the molecular changes that occur in fetal-maternal tissues during placentitis would clarify how this condition leads to abortion, pret...
