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Topic:Immunohistochemistry
Immunohistochemistry (IHC) is a laboratory technique used to visualize specific antigens in tissue sections of horses through the application of antibodies. This method allows for the detailed examination of the distribution and localization of proteins within equine tissues, providing insights into various physiological and pathological processes. IHC is employed in equine research to study a range of conditions, including infectious diseases, inflammatory responses, and neoplastic disorders. By utilizing specific antibodies that bind to target antigens, researchers can identify cellular and tissue changes, contributing to a better understanding of equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of immunohistochemistry in the field of equine research.
Immunolabelling and Micro-Computed Tomography Revealed Age-Related Alterations in 3D Microvasculature of Tendons. Tendon degeneration is common, and its risk increases with age both in humans and horses. Tendon regeneration and healing is limited due to inherent low cell density and vascularisation, and current treatments are insufficient as indicated by scar tissue formation and a high re-injury rate. The tendon vasculature plays a crucial role in tendon homeostasis, regeneration and healing, making it a potential therapeutic target. However, the effect of ageing on the tendon microvasculature is poorly understood. Here, we provide the first comprehensive characterisation of the tendon microvasculature. ...
Abnormal Fetuin-A levels in obese horses are associated with activated TLR4/NF-ƙB/MAPK axis and depleted FBXW7 E3 ubiquitin ligase. Fetuin-A (FetA) is a multifactorial glycoprotein primarily synthesized by the liver with additional expression in adipose tissue, the abundance of which is tightly regulated by the FBXW7 E3 ubiquitin ligase. Recently, FetA has been implicated in the pathogenesis of insulin resistance and associated metabolic failures in humans through its potent and selective inhibition of insulin receptor tyrosine kinase activity, however, no studies have yet directly investigated its role in the development and progression of equine obesity. In this investigation, FetA levels were measured in serum, liver, a...
Corneal Squamous Cell Carcinoma Arising From a Conjunctival Graft Scar in a Horse. To describe the case details of an equine corneal squamous cell carcinoma (SCC) arising from the central scar of a previous conjunctival pedicle graft. Methods: An 18-year-old Thoroughbred gelding, showing a progressive increase in diameter, depth, opacity, and pigmentation of a conjunctival pedicle graft scar of the left eye, 34 months after surgery. Methods: Complete ophthalmic examination, standard histologic and immunohistochemical evaluation of an incisional biopsy to formulate a definitive diagnosis. Carbon dioxide (CO) laser photoablation combined with topical mitomycin C treatment. R...
Endothelial glycocalyx damage is associated with small intestinal disease in horses. To determine if blood and peritoneal fluid levels of syndecan-1, heparan sulfate, and hyaluronan are elevated in horses with small intestinal disease compared to healthy counterparts. Unassigned: 61 horses were categorized into control (n = 20), inflammatory (11), and strangulating (30) groups. Paired peritoneal fluid and blood samples were obtained from each group from December 2016 through December 2024 and analyzed via equine-validated ELISAs for syndecan-1, heparan sulfate, and hyaluronan concentrations. Patient survival to discharge was recorded for all groups. Immunofluorescence staining...
Equine endometrial organoids preserve tissue structure and cycle-stage transcriptional identity†. The endometrium is a hormonally responsive tissue that undergoes cyclic remodeling. Although endometrial organoids have been established in several species, detailed characterization remains limited. Here, we assessed the structural and molecular fidelity of equine endometrial organoids across reproductive cycle stages and during extended culture. Organoids were generated from biopsies collected during estrus and diestrus and analyzed using histology, immunohistochemistry, electron microscopy, and bulk- and single-cell RNA sequencing. Organoids formed polarized cystic structures composed of co...
Proteoglycan 4 is expressed in equine flexor tendons and tendon sheaths, with greater synovial fluid lubricin in tendon sheaths with tendon injury. To describe proteoglycan 4 (PRG4) expression, lubricin tissue distribution, and lubricin synovial fluid concentrations in healthy and injured equine flexor tendons and tenosynovial structures. Unassigned: Synovial fluid and tissues were collected over the span of 1 year. Proteoglycan 4 expression was quantified using quantitative real-time PCR, lubricin synovial fluid concentrations were measured using ELISAs, and lubricin tissue distribution was evaluated with immunohistochemistry. Histology and quantitative real-time PCR data for healthy samples were analyzed using paired t tests, repeated-m...
Quantification of immune cells in full thickness and mucosal biopsies of the duodenum and rectum in a group of slaughter horses. Limited data are available on immune cells in the intestinal wall of healthy horses, hampering interpretation of results in case of disease. Objective: Characterize and quantify the immune cell populations and their distribution in duodenal and rectal biopsies of horses without gastrointestinal disease; compare immune cell counts (ICCTs) between full thickness- and mucosal biopsies. Methods: Twenty horses fit for slaughter, slaughtered for meat production. Methods: Full-thickness and endoscopic forceps obtained mucosal biopsies were taken within 30 min after slaughter from the duodenum and rec...
Immunohistochemical and molecular confirmation of West Nile Virus associated polioencephalomyelitis in a mule from Southern Brazil. West Nile fever is a zoonotic arboviral disease caused by the West Nile Virus (WNV), responsible for deaths in humans, mammals, and birds with associated neurological manifestations. All previous investigations of WNV from Brazil were based primarily on serological and molecular analyses and in humans, equids, and birds in the northern and southeastern regions of the country. This study describes the pathological and molecular findings observed in a mule, from the state of Paraná, southern Brazil, that died during an outbreak involving equids with clinical manifestations of a neurological dis...
MHC compatibility influences the interaction between different types of equine mesenchymal stem/stromal cells and the local immune response. The allogeneic administration of equine mesenchymal stem/stromal cells (MSCs) has numerous advantages over autologous therapy, but their interactions with the patient's immune system need to be further elucidated. These interactions can be influenced by factors such as the compatibility between donor-receptor for the major histocompatibility complex (MHC) and by the MHC expression levels, which can change under different conditions like inflammatory exposure and chondrogeneic differentiation. In this study, we evaluated the local immune response induced by chondrogeneically differentiated (MSC...
[Unilateral leiomyoma in the spermatic cord – a reason for intermittent colic in stallions?]. A 19-year-old Oldenburg stallion was presented multiple times over the course of several months with recurrent colic signs. At each examination, an enlargement of the scrotum was noted, which could not be further specified by palpation and ultrasonographic examination. The increase in circumference was noted both on palpation and on inspection and the size remained unchanged throughout. No special findings were noted during the general examination and the special examination of the colic patient. A few months after the first onset of symptoms, a chicken egg-sized mass was palpated on the right...
Decoding the amniotic membrane transcriptome during equine ascending placentitis. 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...
Cytoplasmic and nuclear Survivin expression in healthy gastric mucosae of equids: A comparative study between horses, donkeys, and mules. Information regarding Survivin protein expression in the gastric mucosa of equids is scarce. This protein has been associated with functions related to modulating apoptosis and promoting mitosis in epithelial cells and is considered part of gastric cytoprotection mechanisms in humans and mice, maintaining mucosal integrity and regulating cellular renewal. Objective: This study aimed to determine the expression of cytoplasmic and nuclear Survivin in gastric mucosae of equids (horses, donkeys, and mules). Methods: A total of 30 healthy stomachs, 10 from each species, were evaluated using histolo...
Melatonin receptors and thyroid stimulating hormone in the equine pars tuberalis: Potential modulators of seasonal reproduction in the mare. The mechanism by which photoperiod influences the hypothalamic-pituitary-gonadal (HPG) axis and regulates seasonal reproduction in horses has yet to be fully elucidated. The hypophyseal pars tuberalis (PT) has been indicated as a critical site for the transduction of melatonin signals through melatonin-responsive, PT-specific cells that produce thyroid-stimulating hormone (TSH) in many mammalian species. However, this has yet to be investigated in horses. The objective of this study was to explore the interaction of melatonin and thyroid-stimulating hormone in the equine HPG axis. Pituitaries ...
Probing Wnt pathway and functional signal in equine melanocytic neoplasms through quantitative proteomics and immunohistochemistry. Equine melanocytic neoplasm (EMN) is a skin tumor commonly observed in grey horses. Limited research has yet to investigate proteomic profiles of EMN, particularly in the early stages and their expression patterns. This study, therefore, aimed to identify signature proteins from tissue biopsies to distinguish early EMN, severe EMN, and normal groups. Results: Using proteomic analysis of 19 tissue samples (normal: n = 6, early EMN: n = 7, severe EMN: n = 6) through LC-MS/MS, 12,310 proteins were identified. Differentially expressed proteins (DEPs) and functional interaction analysi...
Cannabinoid Receptors in the Horse Lateral Nucleus of the Amygdala: A Potential Target for Ameliorating Pain Perception, Stress and Anxiety in Horses. The amygdala is composed of several nuclei, including the lateral nucleus which is the main receiving area for the input from cortical and subcortical brain regions. It mediates fear, anxiety, stress, and pain across species. Evidence suggests that the endocannabinoid system may be a promising target for modulating these processes. Cannabinoid and cannabinoid-related receptors have been identified in the amygdala of rodents, carnivores, and humans, but not in horses. This study aimed to investigate the gene expression of cannabinoid receptors 1 (CB1R) and 2 (CB2R), transient receptor potential...
Gestation-Stage Related Changes in the IGF System Components in the Equine Placenta. The insulin-like growth factor (IGF) system regulates implantation, placental development, and angiogenesis in eutherian mammals. However, little is known about the changes in this system in equine placenta (chorioallantois; CA) and the endometrium (EN) during pregnancy, or the relationship to vascular endothelial growth factor () expression. The current study investigated the expression of the IGF system components, namely the ligands ( and ), their receptors (, , and ), and their binding proteins ( and s) in equine CA at 45 days, 4, 6, 10, and 11 months of gestational age (GA) and immediatel...
Comparative Transcriptome and MicroRNA Profiles of Equine Mesenchymal Stem Cells, Fibroblasts, and Their Extracellular Vesicles. Mesenchymal stem cells (MSCs) are a promising tool in regenerative medicine due to their ability to secrete paracrine factors that modulate tissue repair. Extracellular vesicles (EVs) released by MSCs contain bioactive molecules (e.g., mRNAs, miRNAs, proteins) and play a key role in intercellular communication. This study compared the transcriptomic profiles (mRNA and miRNA) of equine MSCs derived from adipose tissue (AT-MSCs), bone marrow (BM-MSCs), and ovarian fibroblasts (as a differentiated control). Additionally, miRNAs present in EVs secreted by these cells were characterized using nex...
Brain-derived neurotrophic factor and neurotrophic tyrosine receptor kinase-2 in stallion testes: insights into seasonal changes and potential roles in spermatogenesis. Brain-derived neurotrophic factor (BDNF) and its receptor neurotrophic tyrosine receptor kinase-2 (NTRK2) have known important roles in the central nervous system for neurite growth, survival, and differentiation. Nevertheless, the significance of BDNF in spermatogenesis remains unclear in stallions. Therefore, the present study was designed 1) to investigate the expression of BDNF and its receptor NTRK2 and 2) the seasonal variation in the expression patterns of BDNF and NTRK2 in stallions' testes. We used testes from eight postpubertal Thoroughbred stallions collected after a field castratio...
The equine placental extract ameliorates renal damage in mice with adenine-induced chronic kidney disease by inhibiting indoxyl sulfate production. Indoxyl sulfate (IS) is a dietary metabolite of tryptophan that is produced in the liver. It is a uremic toxin that facilitates the progression of chronic kidney disease (CKD). We previously observed that equine placental extract (ePE) inhibited IS synthesis in an inhibition assay using the liver S9 fraction. Unassigned: This study was designed to investigate the effects of ePE on adenine-induced renal failure in mice at the histological and molecular levels to understand the mechanism of action of ePE. Unassigned: We assessed this effect through biochemical and histological analyses using a...
Phylogenetic variation of layer II cortical immature neurons in dog and horse confirms covariance with brain size and neocortical surface. Recent research in brain structural plasticity has identified "immature" or "dormant" neurons in layer II of the cerebral cortex (cortical immature neurons; cINs), cells that remain in a prolonged state of arrested development but retain the ability to resume maturation and integrate functionally into mature cortical circuits. These immature cells are far more abundant in large-brained mammals, being restricted to paleocortex (piriform cortex) in small-brained rodents and extending in the widely expanded neocortical mantle of species with large gyrencephalic brains. In a previous systematic an...
Distribution of endocannabinoid system receptors in the equine hoof: dysregulation as a potential therapeutic target for laminitis. 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 ...
Catalase in Unexpected Places: Revisiting H2O2 Detoxification Pathways in Stallion Spermatozoa. 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...
Development of a two-layer 3D equine endometrial tissue model using genipin-crosslinked collagen scaffolds and 3D printing. 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...
Isolation of SSEA-3-positive mesenchymal stem cells from equine bone marrow and evaluation of their pluripotency. Generally, most mesenchymal stem cells (MSCs) have lower pluripotency and limited differentiation potential than embryonic stem cells (ESCs). However, a small subpopulation of MSCs, called multilineage differentiating stress-enduring (MUSE) cells, exhibit pluripotency. MUSE cells express stage-specific embryonic antigen 3 (SSEA-3), a sphingoglycolipid. Here, we isolated and investigated the pluripotency of SSEA-3-positive MSCs (MUSE cells). Six thoroughbred horses were used as test subjects. MSCs were harvested from the bone marrow of the thoracic vertebrae under ultrasound guidance. Harvested...
Immune cell analysis in equine penile papilloma, in situ squamous cell carcinoma and invasive squamous cell carcinoma: FoxP3+ T regulatory lymphocytes differ according to equine papillomavirus 2 status. Equine penile tumors are common in horses and are often related to infection with equine papillomavirus type 2 (EcPV2). This study investigated the immune cell infiltrate (ICI) of these tumors in horses, focusing on the role of EcPV2. Using multiplex immunohistochemistry (mIHC) for CD3, CD20, and IBA-1 and immunohistochemistry (IHC) for FoxP3, 27 horses with papillomas (5/27), in situ carcinomas (CISs) (3/27), and squamous cell carcinomas (SCCs) (19/27) were evaluated. Eighteen cases tested positive for EcPV2 by either or both in situ hybridization (ISH) and polymerase chain reaction (PCR) (18...
Intracerebral hemorrhage (hemorrhagic stroke) secondary to Bothrops spp. snakebite envenomation in a horse. This report describes the first documented case of intracerebral hemorrhage (ICH) in a horse following Bothrops snakebite envenomation. A 3-year-old mare developed acute kidney injury, systemic hemorrhage, and neurological signs before euthanasia. Postmortem findings confirmed massive ICH with intralesional venom identified by immunohistochemistry. Delayed treatment and insufficient antivenom likely contributed to the outcome. This case highlights the potential for cerebrovascular complications following Bothrops snakebite envenomation in horses, which may present as subtle neurological signs....
Sinonasal angiofibroma in a horse. A 34-year-old Haflinger gelding presented with a unilateral, expansile, intranasal mass that regrew after partial excision. After euthanasia, a large pedunculated mass that originated from the left caudal maxillary sinus and obliterated the left nasal cavity was seen by radiological and macroscopic examination. Histopathology revealed a poorly cellular, expansile, well-vascularized neoplasm composed of a loosely arranged meshwork of spindle cells and collagen fibres. Spindle cells were immunopositive for alpha-smooth muscle actin and occasionally for vimentin, whereas endothelial cells immunol...
Effect of intra-arterial cisplatin on dorsal hoof wall tissue of isolated perfused equine limbs. Equine hoof canker is a chronic proliferative condition of the hoof tissues that is often time-consuming and frustrating to treat. After surgical debridement, topical application of cisplatin (cis-Diamindichlorplatin II) has been reported, requiring repeatedly handling this hazardous substance during bandage changes. Alternatively, intraarterial application of cisplatin could be used, similar to the treatment of some human neoplastic diseases. As the side effects of such a treatment are currently unknown, evaluation of associated risks is necessary before treating live horses. Thus, forelimbs ...
Histological and morphometric changes in cardiac conduction fibers after spontaneous myocardial infarction in horses and dogs. Arrhythmic sudden cardiac death in dogs and horses often results from ventricular arrhythmia secondary to myocardial damage. Despite this, limited data exist on the histomorphometric changes in cardiac conduction fibers (CCFs) and cardiac conduction cells (CCCs) following spontaneous myocardial infarction (MI). This study aimed to characterize morphometric and histological alterations in conduction fibers and their junctions with cardiomyocytes in infarcted hearts of horses and dogs. Unassigned: Ten hearts from horses and 10 from dogs that had died suddenly were examined. Histological and immu...
Evidence of glycan mosaicism in the equine oviduct. We report a lectin histochemical study of oviductal ampulla and isthmus of thirteen mares euthanised at anestrus (1), estrus (5), 1-day post-ovulation (5) or diestrus (2). Staining with a panel of 5 lectins recognizing N-Acetylgalactosamine (GalNAc) residues revealed mosaicism in glycan expression in some specimens of ampulla and isthmus, peaking at estrus and 1-day post-ovulation (ov+1). In the ampulla, this presented as positively stained areas of both ciliated and non-ciliated cells in otherwise negatively-stained ampullae, ranging from small foci (diestrus) to areas up to 200 microns in le...
