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.
Semevolos SA, Brower-Toland BD, Bent SJ, Nixon AJ.Early changes in parathyroid hormone-related peptide (PTH-rP) and Indian hedgehog (Ihh) expression were examined in equine articular osteochondrosis (OC) as a model of a naturally acquired dyschondroplasia. Cartilage was harvested from OC-affected femoropatellar or scapulohumeral joints from immature horses and normal control horses of similar age. PTH-rP expression levels were assessed by semi-quantitative PCR, in situ hybridization, and immunohistochemistry. Ihh protein expression levels were assessed by immunohistochemistry. Elevated PTH-rP protein and mRNA expression were identified in the...
Schwartz AJ, Wilson DA, Keegan KG, Ganjam VK, Sun Y, Weber KT, Zhang J.To determine significant molecular and cellular factors responsible for differences in second-intention healing in thoracic and metacarpal wounds of horses. Methods: 6 adult mixed-breed horses. Methods: A full-thickness skin wound on the metacarpus and another such wound on the pectoral region were created, photographed, and measured, and tissue was harvested from these sites weekly for 4 weeks. Gene expression of type-I collagen, transforming growth factor (TGF)-beta1, matrix metalloproteinase (MMP)-1, and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by quantitative in situ ...
Flaminio MJ, LaCombe V, Kohn CW, Antczak DF.A 12-year-old Quarter Horse mare that was nonresponsive to medical treatment was evaluated for chronic respiratory disease and hepatobiliary disease. Serum immunoglobulin concentrations were measured by use of radial immunodiffusion that revealed trace to nondetectable concentrations of IgG, IgG(T), IgM, and IgA. Use of serum protein electrophoresis confirmed agammaglobulinemia by the absence of the expected peak in the gamma region. In addition, vaccination with tetanus toxoid did not result in specific immunoglobulin production. Flow cytometric analysis of blood lymphocyte subpopulations rev...
Hosaka Y, Kirisawa R, Yamamoto E, Ueda H, Iwai H, Takehana K.Although inflammatory activation of cytokines have been analyzed in various tissues, there have only been a few and as-yet-inconclusive studies on cytokines in equine tendons. In this study, the localizations of 4 cytokines (IL-1alpha, IL-1beta, TNFalpha and IFNgamma) in tendinocytes of the equine superficial digital flexor tendon (SDFT) were analyzed by the use of an immunohistochemical method. In inflamed tendons positive staining for all 4 cytokines antibodies were detected in endotedinieum cells and vascular epithelial cells. In contrast, negative or trace immunoreactions were obtained in ...
Karlström K, Essén-Gustavsson B.An assumed link between red cell hypervolaemia, an excessive amount of training and impaired performance of hypervolaemic horses has led to a theory that the muscle fibres could be affected. Myosin heavy chain (MHC)-based fibre type composition in gluteus medius muscle of red blood cell normo- (NV) and hypervolaemic (HV) Standardbred trotters was evaluated using immunohistochemistry. Muscle biopsies were obtained from 13 NV and 16 HV horses. Serial transverse sections were cut and reacted with antibodies against different isoforms of the myosin heavy chains MHCI, MHCIIA and MHCIIX. Sections we...
Koehler K, Stechele M, Hetzel U, Domingo M, Schönian G, Zahner H, Burkhardt E.This report describes a case of cutaneous leishmaniosis in a horse in southern Germany. Diagnosis is based on histopathology, immunohistochemistry and electron microscopy. The protozoan was identified as Leishmania infantum via PCR and restriction fragment length polymorphism. The horse did not show specific Leishmania antibodies. The lesions healed completely within 6 months without any specific treatment. Since neither the infected horse nor its dam had ever left their rural area, autochthonous infection in Germany cannot be excluded. Factors possibly influencing the epidemiological situatio...
Palaniappan RU, Chang YF, Jusuf SS, Artiushin S, Timoney JF, McDonough SP, Barr SC, Divers TJ, Simpson KW, McDonough PL, Mohammed HO.A clone expressing a novel immunoreactive leptospiral immunoglobulin-like protein A of 130 kDa (LigA) from Leptospira interrogans serovar pomona type kennewicki was isolated by screening a genomic DNA library with serum from a mare that had recently aborted due to leptospiral infection. LigA is encoded by an open reading frame of 3,675 bp, and the deduced amino acid sequence consists of a series of 90-amino-acid tandem repeats. A search of the NCBI database found that homology of the LigA repeat region was limited to an immunoglobulin-like domain of the bacterial intimin binding protein of Esc...
Sato F, Kanno T, Nagasawa S, Yanaihara N, Ishida N, Hasegawa T, Iwanaga T.We investigated the existence of chromogranin A (CgA) in salivary glands of the horse by Western blotting and enzyme immunoassay (EIA) using an antiserum against a peptide sequence of equine CgA. We also compared its cellular distribution between the horse and rat salivary glands with a tyramide signal amplification immunofluorescence technique. Western blotting gave three significant immunoreactive bands (74, 56 and 48 kDa) in adrenal medulla and three major salivary glands of horses. Immunoreactivities for CgA measured by EIA in horses were 154.05 +/- 41.46, 20.32 +/- 5.59 and 4.43 +/- 2.23 ...
Lindner A, Dag S, Marti-Korff S, Quiroz-Rothe E, López Rivero JL, Drommer W.The collection of muscle samples in horses to estimate their performance potential, adaptations to training and diagnosis of exertional myopathies is of increasing interest. This study was designed to examine whether repeated biopsying of the gluteus medius muscle at 7 week intervals alters morphology and muscle fibre variables in the sampling area. The gluteus had been biopsied 3 times previously, each biopsy separated by a 7 week interval. Seven weeks after the last biopsy, this study was initiated. Percutaneous needle biopsies were taken from left and right muscles at 2 depths (2 and 6 cm) ...
Hudson N, Mayhew I, Pearson G.Equine dysautonomia (grass sickness) is a frequently fatal disease of horses characterised by intestinal stasis. Interstitial cells of Cajal (ICC) are the pacemakers and mediators of neurotransmission in the gastrointestinal tract. Impaired ICC-mediated control of motility has been implicated in intestinal disorders in laboratory mammals, humans and in equine grass sickness. The aim of this study was to compare the in vitro electrical properties of ileum from grass sickness cases with horses free from gastrointestinal disease. Intracellular microelectrode recordings were made from smooth muscl...
Budras KD, Schiel C, Mülling CK, Patan B.The preparation of hard tissues such as the equine hoof horn for electron microscopic examination is very difficult. In particular the penetration of fixatives and chemicals used during fixation and embedding is a problem. The objective of this study was to find and implement an alternative method enabling the preparation of high-quality thin sections of hoof horn and other hard tissue, which maintains the hard tissue ultrastructure and can be used for immuno-labeling. Compared to commonly used fixation and embedding techniques, the preparation of thin sections from untreated material method s...
Kreeger JM, Templer A, Tumquist SE, Bailey KL, Johnson PJ, Wilson DA.Paranasal meningioma was diagnosed in a 5-year-old Appaloosa gelding. The mass occupied the right maxillary, frontal, and sphenopalatine sinuses but did not invade the calvarium. The diagnosis was based on histologic evaluation, positive immunohistochemical staining for vimentin and cytokeratin, and ultrastructural features including the presence of interdigitating spindle cells with numerous desmosomes.
Watson ED, Al-Zi'abi MO.The mare is a seasonal breeder and undergoes a period of ovarian transition in spring between winter anoestrus and cyclicity. During spring transition LH concentrations are low and many mares have successive large anovulatory follicular waves which reach the size of preovulatory follicles. Follicular angiogenesis is essential for growth and health of preovulatory follicles. The aim of the present study was to investigate the morphology and vascularity of transitional anovulatory follicles. On gross inspection, the wall of transitional follicles was visibly less well vascularized than that of p...
Deeg CA, Ehrenhofer M, Thurau SR, Reese S, Wildner G, Kaspers B.Equine recurrent uveitis (ERU) is the most serious eye disease in horses worldwide. Despite the fact that ERU is generally considered to be immune mediated, a detailed description of the histopathology of the posterior part of ERU eyes is lacking. Here, we examined sections of paraffin-embedded eyes using histological and immunhistological methods. Twenty seven eyes of 20 horses with ERU and 30 eyes of 15 healthy control horses were included in this study. We could consistently demonstrate an involvement of the retina and the choroid in all examined eyes of horses with spontaneous ERU. In eyes...
Hirayama K, Okamoto M, Sako T, Kihara K, Okai K, Taharaguchi S, Yoshino T, Taniyama H.Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific en...
Lorenzo PL, Liu IK, Carneiro GF, Conley AJ, Enders AC.Epidermal growth factor (EGF) has been shown to have a positive effect during oocyte in vitro maturation in several species. This study was performed to establish the capacity of equine oocytes to undergo nuclear maturation in the presence of EGF and to localise its receptor in the equine ovary by immunohistochemical methods. Oocytes were obtained by aspiration and subsequent scraping from equine follicles (15-25 mm diameter) and cultured in 3 different treatment groups for 36 h: control Group (modified TCM 199 with 0.003% BSA), EGF Group (TCM-199 supplemented with 50 ng/ml EGF) and EMS Group ...
Henson KL, Alleman AR, Kelley LC, Mahaffey EA.Immunohistochemical techniques were used to examine 29 cases of equine lymphoma for estrogen receptor (ER) and progesterone receptor (PR) expression. The lymphomas examined included T-cell-rich large B-cell lymphomas, B-cell neoplasms, and T-cell lymphomas. The individual cases were also classified according to the anatomic location of the tumors. One normal equine lymph node was also examined for ER and PR expression. All of the cases of equine lymphoma and the normal lymph node were negative for Er. A total of 16/29 (55%) PR-positive lymphomas were identified. Seven of the 12 (58%) T-cell-ri...
Klomkleaw W, Kasashima Y, Fuller GA, Kobayashi A, Yoshihara T, Oikawa MA, Izumisawa Y, Yamaguchi M.An anatomical study of horse lumbrical muscle (Lm) was carried out by light and electron microscopy in combination with immunochemical and cytochemical methods. Paraffin sections were subjected to haematoxylin and eosin (H & E) and Masson's trichrome staining for morphometric analysis. Paraffin sections were also used for immunostaining by anti-PGP 9.5 for reaction with nerve-protein associated-structures, anti-heat-shock protein 70 (hsp 70) for detection of gene expression changes, anti-fast myosin for the determination of muscle fibre types, and for detection of apoptotic gene expression...
Gray AW, Davies ME, Jeffcott LB.Cathepsin K and cathepsin B were immunolocalised in equine osteoclasts (OC s) present in ex vivo cartilage/subchondral bone samples. Samples were obtained post mortem from the lateral trochlear ridge (LTR) of six horses and ponies aged between 303 days gestation to 8 months. Strong expression of cathepsin K was detected in OC s, particularly those located at the osteochondral junction, apparently involved in the resorption of calcified cartilage. Cathepsin K expression was also detected in hypertrophic chondrocytes and in the endothelial cells of some blood vessels penetrating the hypertrophic...
Sanchez B, Nieto A, de Ruiz Leon MA, Rodríguez J, Flores J.A lymphangiosarcoma with metastases was found in a horse that presented with respiratory distress and edema in the ventral thorax and abdomen. The necropsy revealed diffuse edema in the subcutaneous connective tissue. Mediastinal, mesenteric, iliac, and renal lymph nodes were enlarged and white with soft, yellowish necrotic areas. Histologic examination revealed numerous channels and disorganized vessels lined by large polyhedral, polymorphic cells. Tumor metastases were observed in the spleen, lungs, and kidneys. Immunohistochemical evaluation of the tumor cells demonstrated positive staining...
Theoret CL, Barber SM, Gordon JR.To describe the localization of immunoreactive transforming growth factor (TGF)-beta1 in both normal skin and full-thickness dermal wounds of the limb and the thorax of the horse. Methods: Six full-thickness excisional wounds were created on the lateral aspect of one metacarpal region and on the midthoracic area of each horse. Sequentially collected tissue specimens from wound margins were assessed for TGF-beta1 expression by immunohistochemistry. Methods: Four horses (2 to 4 years of age). Methods: A neutralizing monoclonal anti-human TGF-beta1 antibody was used to detect the spatial expressi...
Pinkerton ME, Bailey KL, Thomas KK, Goetz TE, Valli VE.A 17-year-old Quarterhorse gelding with a clinical diagnosis of protein-losing enteropathy was submitted for necropsy following a 4-5-month duration of weight loss, decreased appetite, and hypoproteinemia. Gross findings included multiple 1-2-cm diameter ulcers on the luminal surfaces of the duodenum and ileum. Histologic examination revealed individual large, round cells infiltrating much of the mucosal epithelium of the duodenum, jejunum, ileum, and colon in addition to multifocal areas of ulceration. Similar round cells infiltrated Brunner's glands and expanded the submucosa beneath the foc...
Fortier LA, Mohammed HO, Lust G, Nixon AJ.Composites of chondrocytes and polymerised fibrin were supplemented with insulin-like growth factor-I (IGF-I) during the arthroscopic repair of full-thickness cartilage defects in a model of extensive loss of cartilage in horses. Repairs facilitated with IGF-I and chondrocyte-fibrin composites, or control defects treated with chondrocyte-fibrin composites alone, were compared before death by the clinical appearance and repeated analysis of synovial fluid, and at termination eight months after surgery by tissue morphology, collagen typing, and biochemical assays. The structure of cartilage was ...
Dingboom EG, van Oudheusden H, Eizema K, Weijs WA.In order to obtain broader insights into the equine musculoskeletal system, we studied the fibre type composition of 2 locomotory muscles in biopsies from Dutch Warmblood foals taken at 3 different ages in the first year postpartum. The muscle fibre types were determined histochemically as well as immunohistochemically. ATPase-characterised IIB fibres appear to express either IId or type lIa plus IId myosin heavy chain (MHC). A high percentage of fibres classified as IIA with ATPase expressed both fast types of MHC. The type I classification by the 2 methods matched almost completely. There wa...
Dahm AM, de Bruin A, Linat A, von Tscharner C, Wyder M, Suter MM.We describe the establishment and characterisation of equine keratinocyte cultures with maintenance of a high proliferative capacity up to the second passage. Improved attachment and growth were obtained by seeding primary cells on equine feeder layers. Subcultured keratinocytes showed optimal growth when seeded on collagen type I. The proliferation rate of cells on this substrate exceeded that seen for cells seeded on equine feeder layers. By immunohistochemistry, epithelial origin and state of differentiation of the equine keratinocytes were determined. They expressed keratin and desmoplakin...
Marsh AE, Hyun C, Barr BC, Tindall R, Lakritz J.Equine protozoal myeloencephalitis (EPM), caused by a protozoal parasite infection of the central nervous system, is the most commonly diagnosed neurologic disease of horses in North America. In specific regions of the United States approximately 50% of the horse population is seropositive to Sarcocystis neurona. However, not all seropositive horses develop clinical signs. Detailed clinical examination, along with cerebrospinal fluid antibody evaluation are often used to diagnose EPM. Postmortem evaluation of the brain stem and spinal cord for histopathologic lesions compatible with nonsuppura...
Dixon S, Haswell M, Harrington D, Sutcliffe IC.We have investigated the surface localisation of the phosphotransferase system protein HPr in the equine pathogen Streptococcus equi subsp. equi using immunogold localisation and transmission electron microscopy. Like the LppC acid phosphatase lipoprotein, a reference surface antigen, the S. equi HPR could be clearly detected on the surfaces of intact cells. This study is consistent with previous reports that some streptococcal HPr is cell surface associated and suggests that the extracytoplasmic mobilisation and transfer of phosphate groups by streptococci warrant further investigation.
Tanaka Y, Taniyama H, Tsunoda N, Shinbo H, Nagamine N, Nambo Y, Nagata S, Watanabe G, Herath CB, Groome NP, Taya K.Immunolocalization of the inhibin (a) and inhibin/activin (beta3A and betaB) subunit proteins in equine fetal testes was investigated to determine the ability of the fetal testis to produce inhibins at 120, 150, 200, and 250 days of gestation. In addition, concentrations of immunoreactive (ir)-inhibin, inhibin pro-aC, and inhibin A in both the maternal and fetal circulation were measured. It was found that plasma concentrations of ir-inhibin, inhibin pro-alphaC, and inhibin A were much higher (P < .05) in the fetal than in the maternal circulation at any stage of gestation examined. Similar...
Szeredi L, Makrai L, Dénes B.The first objective of this study was to develop an immunohistochemical procedure for rapid detection of Rhodococcus equi in impression smears from affected organs of foals on postmortem examination. The second aim was to demonstrate whether R. equi can be detected in smears of tracheal exudates collected from the same foals using an immunohistochemical method. Impression smears and cryostat and paraffin-embedded sections were made from the lungs and mediastinal lymph nodes of three foals (A, B and C) that had died of respiratory disease caused by R. equi, and also from the caudal mesenteric l...
Van Wulfen KK, Bowker RM.To localize substance P (SP) and neurokinin A (NKA) and their receptors in the insertion of the distal sesamoidean impar ligament (DSIL), deep digital flexor tendon (DDFT), and dorsal hoof wall of healthy feet of horses. Methods: 18 healthy feet from horses. Methods: Samples from the dorsal hoof wall and insertion of the DSIL and DDFT of 10 feet were processed for immunocytochemical analysis, using rabbit polyclonal antisera raised against SP and NKA. Tissue sections from 8 feet were incubated with 125-labeled SP to localize tachykinin receptors and their specificity and with control solutions...
Tanaka Y, Nagamine N, Nambo Y, Nagata S, Nagaoka K, Tsunoda N, Taniyama H, Yoshihara T, Oikawa M, Watanabe G, Taya K.In mares, circulating immunoreactive inhibin concentrations increase during the follicular phase and decrease at the start of the LH surge. Thereafter, sharp increases in circulating immunoreactive inhibin concentrations, the 'ovulatory increase', are observed during ovulation. In the present study, the cellular sources and molecular form of ovarian inhibin were investigated to determine the mechanism responsible for this unique ovulatory increase. Three sizes of ovarian follicles (small, 30 mm in diameter) were selected. Inhibin alpha-subunit was localized by immunohistochemistry to the gran...
Bulak K, Łopuszyński W, Lutnicki K, Pomorska-Zniszczyńska A, Śmiech A, Jodłowska-Jędrych B.Granular cell tumor (GCT) is a soft tissue neoplasm characterized by abundant intracellular eosinophilic granules. The majority of GCTs are benign, although some display malignant behavior. Furthermore, GCTs may mimic other neoplasms. The clinical course and biology of GCTs are poorly understood. Regarding the histogenesis of GCT, a Schwann cell origin is currently favored in light of immunohistochemical and ultrastructural analyses. However, based on literature data, some of the primitive GCTs show non-neural origin; therefore, the histogenesis of this tumor has remained enigmatic. Granular c...
Sikora M, Nowak M, Racheniuk H, Wojtysiak K, Kozdrowski R.Endometritis and endometrosis have been and still are the major reasons for infertility in mares. The diagnosis of endometritis can be based on cytology and microbiology, but endometrial biopsy is still the only way to diagnose endometrosis in the mare. Our study attempted to determine if a single biopsy using his-topathology and immunohistochemistry is sufficient to ascertain reasons for infertility in Icelandic mares. The objectives of this study were to examine the relationship between deviations in endometrial biopsies in terms of prostaglandin-endoperoxide synthase-2 (PTGS-2) and fibronec...
Morini M, Peli A, Rinnovati R, Magazzù G, Romagnoli N, Spadari A, Pietra M.Severe equine asthma (EA) syndrome is a chronic obstructive disease characterized by exaggerated contraction, inflammation, and structural alteration of the airways in adult horses, when exposed to airborne molds and particulate material. However, little is known about the relationship between the degree and type of inflammation on one hand, and the severity of the disease and the response to treatment on the other. Furthermore, to date, very few studies evaluate the diagnostic value of histology and immunohistochemical features of endoscopic biopsies on subjects with severe equine asthma. To ...
Furness MC, Bienzle D, Caswell JL, Delay J, Viel L.Recurrent airway obstruction (RAO) in the horse is a disease characterized by reversible bronchoconstriction and by mucus and neutrophil accumulation in the airways. It has been hypothesized that in horses with RAO, remodeling changes occur that are similar to those described in humans with asthma. Although collagen fibrils are present surrounding normal airways, they are a prominent feature of airway remodeling in human asthma with evidence of enhanced collagen III and I fibril deposition. An immunolabeling method was developed to identify collagen I and III in equine lung and to describe the...
Miller LMJ, Woodward EM, Campos JR, Squires EL, Troedsson MHT.Sperm protein at 22 kDa has been associated with fertility. Objective: The objectives of this study were to determine (1) the localization pattern of SP22 on ejaculated and caudal epididymal equine spermatozoa and in epididymal fluid, and to (2) characterize SP22 protein and mRNA expression in testicular and epididymal tissues in response to heat-induced testicular degeneration. Methods: Semen was collected before and after hemi-castration, as well as prior to and following insulation of the remaining testes, and tissue specimens were collected for analysis. Results: Histopathology confirmed ...
Wilsher S, Lefranc AC, Allen WR.Fillies completely devoid of endometrial glands (uterine gland knockout; UGKO) would make ideal experimental models in which to study the role of endometrial histotroph in embryogenesis and early fetal development in the mare. Objective: Administration of a synthetic progestagen plus oestrogen to newborn filly foals and, thereafter, at regular intervals to age 6 months, would permanently suppress endometrial gland development. Methods: Nine half-sister Thoroughbred filly foals were treated, in 3 groups, with: A) the weakly active progestagen, norgestomet, administered from birth to age 6 month...
Meulenbroeks C, van der Meide NM, Zaiss DM, van Oldruitenborgh-Oosterbaan MM, van der Lugt JJ, Smak J, Rutten VP, Willemse T.Insect bite hypersensitivity (IBH) in horses is a seasonal, IgE-mediated, pruritic skin disorder primarily caused by Culicoides spp. We hypothesize that a mixed Th2/Th1-type immune status, off season, alters into Th2-dominated immune reactivity in the skin of IBH-affected ponies in the IBH season. To study these immune response patterns Culicoides-specific IgE levels, skin histopathology and cytokine and transcription factor mRNA expression (IL4, IL10, IL13, IFNγ, FoxP3 and CD3(ζ)) in lesional and non-lesional skin of ponies affected by IBH in the IBH season were compared with those of the s...
Schnabel CL, Steinig P, Koy M, Schuberth HJ, Juhls C, Oswald D, Wittig B, Willenbrock S, Murua Escobar H, Pfarrer C, Wagner B, Jaehnig P, Moritz A....Deoxyribonucleic acid (DNA) vaccines are used for experimental immunotherapy of equine melanoma. The injection of complexed linear DNA encoding interleukin (IL)-12/IL-18 induced partial tumour remission in a clinical study including 27 grey horses. To date, the detailed mechanism of the anti-tumour effect of this treatment is unknown. Results: In the present study, the clinical and cellular responses of 24 healthy horses were monitored over 72 h after simultaneous intradermal and intramuscular application of equine IL-12/IL-18 DNA (complexed with a transfection reagent) or comparative substanc...
Pavone S, Mandara MT.The interstitial cells of Cajal (ICC) play a key role in the control of intestinal motility and have been implicated in several human gastrointestinal dysmotility syndromes, in equine grass sickness and in other intestinal disorders where a significant reduction in ICC density was observed. Objective: To investigate the density of ICC in clinically normal horses, ICC c-Kit expression was evaluated by image analysis in order to obtain numerical data. Methods: Intestinal samples from the jejunum to small colon from 5 clinically normal horses were studied. Immunohistochemical labelling of ICC was...
Fernandes CB, Loux SC, Scoggin KE, Squires EL, Troedsson MH, Esteller-Vico A, Ball BA.The cervix is a dynamic structure that undergoes dramatic changes during the estrous cycle, pregnancy and parturition. It is well established that hormonal changes, including estrogens, progestogens and prostaglandins, regulate the expression of key proteins involved in cervical function. The arachidonic acid cascade is important in the remodeling and relaxation of the cervix in the days preceding parturition. Despite the complexity of this mechanism, regulation of cervical function has received little study in the mare. Therefore, the objective of this study was to compare the expression of e...
Brocca G, Centelleghe C, Padoan E, Stoppini R, Giudice C, Castagnaro M, Zappulli V.A 24-year-old Irish Cob mare was presented with a peripheral iris mass, which was surgically resected and diagnosed as an undifferentiated neuroepithelial tumor. A few months later, a relapse occurred with histological features characterized by a more solid appearance and squamous differentiation. Subsequently, the mare was presented with rapidly spreading multiple subcutaneous masses and, at the onset of neurological signs, was humanely euthanized and subjected to a complete post mortem examination. The necropsy confirmed the presence of numerous widespread masses in the subcutaneous tissue, ...
Townsend J, Westcott K, Tortonese DJ.In the horse, pronounced changes in fertility occur annually in response to photoperiod. However, the mechanisms regulating gonadotrophin synthesis and release in this species remain unclear. Here, we investigated the expression of gonadotrophin subunits and GnRH receptor (GnRH-R) mRNA in the pituitary glands of Thoroughbred horses during the breeding (BS) and non-breeding (NBS) season. Seasonal effects on the prevalence of gonadotrophs in the pars distalis were also examined. GnRH-R and common alpha-, LHbeta- and FSHbeta-subunit mRNA contents were determined by Northern analysis and the preva...
Woodward MC, Andrews FM, Kearney MT, Del Piero F, Hammerberg B, Pucheu-Haston CM.To characterize the response of skin of nonallergic horses following ID injection of polyclonal rabbit anti-canine IgE (anti-IgE) and rabbit IgG. Methods: 6 healthy horses. Methods: Skin in the cervical area was injected ID with anti-IgE and IgG. Wheal measurements and skin biopsy specimens were obtained before and 20 minutes and 6, 24, and 48 hours after injection. Tissue sections were evaluated for inflammatory cells at 4 dermal depths. Immunohistochemical analysis for CD3, CD4, and CD8 was performed, and cell counts were evaluated. Results: Anti-IgE wheals were significantly larger than IgG...
Visser MB, Pollitt CC.The basement membrane (BM) is a thin layer of extracellular matrix that regulates cell functions as well as providing support to tissues of the body. Primary components of the BM of epithelial tissues are laminin-332 (Ln-332) and collagen type IV. Equine laminitis is a disease characterized by destruction and dislocation of the hoof lamellar BM. Immunohistochemistry was used to characterize the distribution of Ln-332 and collagen type IV in the organs of normal horses and these proteins were found to be widespread. Analysis of a panel of tissue samples from horses with experimentally-induced l...
Marsh AE, Chaney SB, Howe DK, Saville WJ, Reed SM.Several reports indicate the presence of small tissue cysts associated with Sarcocystis neurona infections. Several failed attempts to develop tissue cysts in potential intermediate host using in vitro derived parasites originally isolated from horses with equine protozoal myeloencephalitis suggest that the experimental methods to achieve bradyzoites with those isolates was not possible. Those prior studies reported the lack of detectable sarcocysts based on histology and in vivo feeding trials. A recent report of successful production and detection of small sarcocysts triggered us to review a...
Watanabe T, Hosaka Y, Yamamoto E, Ueda H, Tangkawattana P, Takehana K.The Golgi tendon organ (GTO) is an encapsulated fusiform mechanoreceptor siding in the musculo-tendinous junction of many animal species. Inhibitory function of afferent nerve fibers distributed from nearby motor units, the organ responds to active tension exerted onto the muscle. The morphological features of the equine GTO have not yet been elucidated. Additionally, there is some controversy regarding to the existence of the GTO in the equine superficial digital flexor tendon (SDFT). Therefore, immunohistochemistry and immunoelectron microscopy using alcian blue (pH 2.5) staining and the sil...
Moore PF, Hacker DV, Buyukmihci NC.Angiosarcomas arising in ocular tissues of four aging horses are described. Tumors were locally invasive and eventually metastasized via the mandibular and cervical lymph nodes. Pathologically, the tumors contained well-differentiated regions, in which vascular channels were lined by pleomorphic endothelial cells, as well as poorly-differentiated regions, in which vascular channels were either rudimentary or absent. Red blood cells were scare in vascular structures formed by the tumors. Factor VIII related antigen (VIII:RAg), a blood vascular endothelial marker, was demonstrated by immunohisto...
Lançoni R, Celeghini ECC, Gonella-Diaza AM, Júnior VG, de Carvalho CPT, Zoca GB, Garcia-Oliveros LN, Batissaco L, Oliveira LZ, de Arruda RP.This study aimed to assess the semen ubiquitin levels of stallions with good (GF) and poor semen freezability (PF) and to evaluate the relationship between sperm ubiquitination and sperm morphological defects. Five ejaculates from eight adult stallions (n = 40) were collected and cryopreserved. Then, the ubiquitin level in equine sperm cells was assessed by immunohistochemistry with epifluorescence microscopy, and sperm morphology was assessed by differential interference contrast microscopy. Sperm cells were classified according to the intensity (classification 1: from I to IV; I = very l...
Diel de Amorim M, Klein C, Foster R, Dong L, Lopez-Rodriguez MF, Card C.Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is an enzyme that metabolizes oxytocin in serum and tissues. The presence of oxytocin/neurophysin I (OXT), oxytocin and LNPEP and their relationship to other genes is unknown in the equine conceptus. Our objective was to characterize gene expression of LNPEP and OXT on D8, 10, 12, 14, 15, 16 and 21 conceptuses in relationship to other genes. Immunohistochemistry, western blot and liquid chromatography with tandem mass spectrometry (LC-MS/MS) were used for identification of oxytocin and LNPEP in D15, 16 and 18 conceptuses. LNPEP was increas...
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...
Ekfalck A, Appelgren LE, Funkquist B, Jones B, Obel N.The distribution of 35-S-labelled cysteine and methionine in the epidermis of the equine hoof following 2 hours of intra-arterial injection was studied by microautoradiography. Material for autoradiography was obtained by biopsy about 1 hour after termination of the intra-arterial injection and also 10 and 40 days later. In the specimens obtained one hour after the injection of labelled cysteine and methionine, the amount of radioactivity in the matrix and in the most proximal part of the laminar layer was very high. There was a clear difference between the distribution of the two labelled ami...
Roessner HA, Hurcombe SD, Klein CE, Hopster K, Engiles JB.To compare end-to-end jejunal anastomoses with a one-layer (Utrecht) and two-layer (simple continuous/Cushing) patterns. Methods: Experimental study. Methods: Eight healthy adult horses. Methods: Jejunal end-to-end anastomoses were performed in randomly assigned one-layer or two-layer patterns. Horses were recovered from surgery and monitored for complications. At 14 days, the opposite pattern was performed (cross-over design) prior to euthanasia. Duration of closures was compared between patterns. Serosal width was measured before harvesting anastomotic sites from the first procedure. Lumin...
Grau-Roma L, Peckham R, Paton J, Stahel A, de Brot S.The postmortem examination of a 14-y-old Appaloosa gelding with clinically diagnosed pituitary pars intermedia dysfunction showed a unique finding of moderate multifocal lymphocytic hypophysitis (LH). The pituitary glands of 24 horses submitted for postmortem examination were examined grossly and examined histologically for the presence of lymphocytes. Of these 23 horses, 1 additional case suffered from moderate LH. The 2 cases with LH tested negative for Equid herpesvirus 1 and 4 by polymerase chain reaction and immunohistochemistry (IHC), and no viral particles were observed by electron micr...
Hallamaa RE, Saario E, Tallberg T.Healing sarcoids were followed in 18 horses which had taken part in previous clinical studies on a total of 29 horses suffering from either primary or recurrent sarcoids, treated with bio-immunotherapy. In the present study, attention was paid to changes observed in these fibroblastic skin tumours during their regression. The tumours were surgically debulked leaving the base in the skin. The horses were immunized according to bio-immunotherapy at 2- to 4-week intervals with an autogenous vaccine made from the excised part of the tumour until the base had visibly regressed. Healing was followed...
Abu-Seida AM, Wafy MN, Hassan EA, Ahmed KA.Ocular neoplasms represent 10% of all equine neoplasms and ocular squamous cell carcinoma (OSCC) is the most common one. This report describes the clinical presentation, laboratory, ultrasonography, postmortem, histopathology and immunohistochemistry findings of a mixed ocular neoplasm in a 10-year-old draft mare. The mare had a one-year history of left ocular mass. Complete destruction of the left eye with loss of vision was observed. A large ulcerated mass oozing blood replaced the left eye. Left parotid lymph node was swollen and had a sinus tract discharging pus. Ultrasonography revealed a...
Sasaki K, Igarashi S, Amasaki T, Amasaki H, Nishita T, Kano Y, Asari M.Immunohistochemical localizations of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in equine and bovine digestive tracts were studied. In the horse, epithelial cells in both the oesophagus and non-glandular part of the stomach lacked all three isozymes. In contrast, surface epithelial and parietal cells in the glandular region of the stomach showed reactivity for CA-II. In the small intestine, absorptive columnar cells covering the villi in the duodenum were positive for CA-II. The epithelium of the jejunum and ileum lacked all three isozymes. In the large intestine, CA-II was detected ...
Beccati F, Pepe M, Pascucci L, Ceccarelli P, Chiaradia E, Mancini F, Mandara MT.The purpose of this study was to delineate the pattern of sympathetic innervation in the suprasesamoidean region of the deep digital flexor tendon (DDFT) in horses using immunohistochemical staining (IHC) for tyrosine hydroxylase (TH) and alpha-1 adrenergic receptor (α1-AR). Fourteen forelimbs were collected from 10 horses. Longitudinal sections of the suprasesamoidean region of healthy DDFTs were harvested. Most of the sympathetic innervation was found to be in the walls of blood vessels. The tendon tissue proper was sparsely innervated, with a lesser degree of innervation within the dorsal ...
Gondim FJ, Modolo LV, Campos GE, Salgado I.Mammalian skeletal muscle expresses splice variants of neuronal nitric oxide synthase (nNOS). Skeletal muscles have a metabolically heterogeneous population of myofibers, and fiber composition in equine skeletal muscle is correlated with athletic ability in endurance events. In this study, we investigated whether nNOS expression in equine skeletal muscle is related to fiber type and endurance training. Biopsy samples obtained from the gluteus medius of sedentary- (SH) and endurance-trained (TH) horses were examined for the electrophoretic mobility of myosin heavy chain (MHC) and NOS activity. ...
Dall'Aglio C, Bazzucchi C, Mercati F, Ceccarelli P.Leptin is a hormone widely diffused in the mammalian body in which it plays functions that go far beyond control of appetite and energy metabolism. The finding that it is present in the major salivary glands of various animal species is of interest for the functional implications that it may imply. Since there are no data on the presence of leptin and its receptor in the minor salivary glands, the aim of this study was to demonstrate their presence and distribution in such glands of donkeys. This latter was chosen as species of reference because the monogastric herbivore shows intense salivati...
