The endothelium in horses is a thin layer of cells lining the interior surface of blood vessels, playing a significant role in vascular function and homeostasis. It is involved in the regulation of blood flow, coagulation, and immune response. The endothelium also facilitates the exchange of substances between the bloodstream and surrounding tissues. In horses, endothelial function can be affected by various factors, including inflammation, metabolic disorders, and infectious diseases. Understanding the behavior and pathology of the endothelium is essential for comprehending equine cardiovascular health and disease mechanisms. This page gathers peer-reviewed research studies and scholarly articles that explore the structure, function, and pathological changes of the endothelium in horses.
Berhane Y, Bailey SR, Putignano C, Elliott J.The role of endothelium-derived relaxing factors was studied in the regulation of vascular responses in the Krebs perfused equine isolated digit. Perfusion pressure was recorded in response to bolus doses of 5-hydroxytryptamine (6 nmol) alone or co-administered with carbachol (CCh; 0.2 micromol), bradykinin (BK; 0.2 nmol), substance P (SP; 0.2 nmol) or sodium nitroprusside (SNP; 0.2 micromol). N(omega)-Nitro-L-Arginine methyl ester hydrochloride (L-NAME; 300 microm) caused partial but significant inhibition of CCh-induced vasodilatory response, whereas BK and SP-induced responses were resistan...
de la Rebière G, Franck T, Deby-Dupont G, Salciccia A, Grulke S, Péters F, Serteyn D.As heparins are sometimes used to prevent equine laminitis, the interactions between equine neutrophil myeloperoxidase (MPO), unfractionated (UFH) and fractionated low molecular weight (LMWH) heparins and digital endothelium have been investigated. The effects of the heparins on purified equine MPO activity were tested by immunocapture followed by enzymatic detection. Endothelium-MPO interactions were assessed by measuring total and active MPO uptake by arterial and venous digital endothelial cells in culture with or without the addition of heparins. A dose-dependent MPO inhibition by UFH and ...
Allen WR, Gower S, Wilsher S.Polyclonal antisera for vascular endothelial growth factor (VEGF) and its two main receptor molecules, VEGF-I (Flt) and VEGF-II (KDR), were used in a conventional immunocytochemical staining method to localize these angiogenic ligand molecules in the endometrium and placenta of the mare during the oestrous cycle and pregnancy. The anti-VEGF and anti-Flt sera both labelled the lumenal and glandular epithelia of the endometrium throughout the oestrous cycle and both the invasive trophoblast cells of the endometrial cups and the non-invasive trophoblast of the allantochorion in pregnancy. The ant...
Robertson TP, Moore JN, Noschka E, Lewis TH, Lewis SJ, Peroni JF.To determine the effects of inhibition of Rho-kinase or Src-family protein tyrosine kinases (srcPTK) on agonist-induced contractile responses in equine laminar arteries and veins. Methods: Laminar arteries and veins obtained from 13 adult mixed-breed horses. Methods: Laminar vessels were mounted on myographs and exposed to phenylephrine (PE), 5-hydroxytryptamine (5-HT), prostaglandin F(2) (PGF(2)), and endothelin-1 (ET-1) with or without the Rho-kinase inhibitor Y-27632 (10 microM), srcPTK inhibitor PP2 (10 microM), or a negative control analogue for PP2 (PP3; 10 microM). Results: Responses to...
Huang H, Lavoie-Lamoureux A, Moran K, Lavoie JP.Little is known concerning the possible contribution of T helper 2 (Th2)-type cytokines to the recruitment of neutrophils into the lung tissue. In the present study, endothelial cells from equine pulmonary arteries were cultured in the presence of recombinant equine (re) IL-4 and reIL-5, and the cytokine mRNA expression of molecules implicated in the chemotaxis and migration of neutrophils was studied using real-time RT-PCR. The functional response of reIL-4-induced endothelial cell stimulation on neutrophil migration was also studied using a chemotaxis chamber. ReIL-4 either increased the exp...
Padilla DJ, Epp TS, McDonough P, Marlin DJ, Erickson HH, Poole DC.During high intensity exercise, the very high pulmonary artery pressure (Ppa) experienced by Thoroughbred horses is considered a major factor in the aetiology of exercise-induced pulmonary haemorrhage (EIPH). Recently, endothelin-1 (ET-1), a potent vasoconstrictive hormone, has been found to increase Ppa in horses at rest via binding to its ET-1A receptor subtype. In addition, plasma concentrations of ET-1 are increased in horses during and after high intensity exercise. Objective: If ET-1 increases Ppa during exercise in the horse, administration of a specific ET-1A antagonist would decrease ...
Eades SC, Stokes AM, Johnson PJ, LeBlanc CJ, Ganjam VK, Buff PR, Moore RM.To quantify changes in endothelium-derived factors and relate those changes to various aspects of digital hemodynamics during the prodromal stages of carbohydrate overload (CHO)-induced laminitis in horses. Methods: 20 adult horses without abnormalities of the digit. Methods: Digital and jugular venous blood samples were collected at 1-hour intervals (for assessment of endothelin-1 [ET-1] immunoreactivity and measurement of glucose, insulin, and nitric oxide [NO] concentrations) or 4-hour intervals (CBC and platelet-neutrophil aggregate assessment) for 8 hours or 16 hours after induction of CH...
Berhane Y, Elliott J, Bailey SR.Haemodynamic disturbances leading to ischaemia and reperfusion injury of the digit are thought to be involved in the pathophysiology of acute equine laminitis. Identification of physiological regulators of blood flow through the equine digit is important in identifying factors, which may predispose animals to laminitis. A method was developed to assess endothelium-dependent responses of the isolated Krebs-perfused equine digit by co-administration of 5-hydroxytryptamine (5-HT) with vasodilator agents, carbachol (CCh), bradykinin (BK) and substance P (SP). Bolus co-administration of CCh (0.02-2...
Stokes AM, Venugopal CS, Hosgood G, Eades SC, Moore RM.The goals of this study were to determine the concentration-response (C-R) relationship of endothelin-1 (ET-1), compare 2 ET-receptor antagonists and determine the antagonist concentrations that block the vasomotor effects of ET-1, and compare the effectiveness of ET-1 and previously studied vasoconstrictors in equine palmar digital arterial and venous rings in vitro. Vessel rings from 8 nonlaminitic horses were placed in Tyrode's solution, 1 side fixed to the floor of an organ bath and the other side fixed to a force-displacement transducer. Two separate studies were conducted: (I) incubation...
Dubuc V, Lepault E, Theoret CL.Wound repair in horse limbs is often complicated by excessive fibroplasia and scarring. Occlusion of the microvessels populating the granulation tissue appears to be involved in the excessive accumulation of extracellular matrix during the repair of limb wounds. This study aimed to determine whether endothelial cell hypertrophy or hyperplasia, or both, contribute to microvascular occlusion and whether the pericyte is involved in this anomaly. We created 5 wounds, each 2.5 x 2.5 cm, on both forelimbs and on the body of 6 horses. One limb was bandaged to stimulate excessive wound fibroplasia. We...
Polikepahad S, Paulsen DB, Moore RM, Costa LR, Venugopal CS.To immunohistochemically determine the expression of endothelin (ET) receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD). Methods: Tissue specimens obtained from 8 healthy and 8 SPAOPD-affected horses. Methods: Horses were examined and assigned to healthy and SPAOPD groups. Horses were then euthanatized, and tissue specimens containing bronchi of approximately 4 to 8 mm in diameter were immediately collected from all lung lobes, fixed in zinc-formalin solution for 12 hours, and embedded in...
Hasebe R, Kimura T, Nakamura K, Ochiai K, Okazaki K, Wada R, Umemura T.Equine herpesvirus 1 (EHV-1) shows endotheliotropism in the central nervous system (CNS) of infected horses. However, infection of endothelial cells has not been observed in the CNS of infected mice. To explore the basis for this difference in endotheliotropism, we compared the susceptibility of equine brain microvascular endothelial cells (EBMECs) and mouse brain microvascular endothelial cells (MBMECs) to EHV-1 infection. The kinetics of viral growth in EBMECs was typical of a fully productive infection whereas viral infection in MBMECs seemed to be nonproductive. Immunofluorescence microsco...
Brindley MA, Maury W.Recently, it has become evident that entry of some retroviruses into host cells is dependent upon a vesicle-localized, low-pH step. The entry mechanism of equine infectious anemia virus (EIAV) has yet to be examined. Here, we demonstrate that wild-type strains of EIAV require a low-pH step for productive entry. Lysosomotropic agents that inhibit the acidification of internal vesicles inhibited productive entry of EIAV. The presence of ammonium chloride (30 mM), monensin (30 microM), or bafilomycin A (50 nM) in the medium dramatically decreased the number of EIAV antigen-positive cells. We foun...
Peroni JF, Moore JN, Noschka E, Grafton ME, Aceves-Avila M, Lewis SJ, Robertson TP.Equine laminitis is a crippling condition associated with a variety of systemic diseases. Although it is apparent that the prodromal stages of laminitis involve microvascular dysfunction, little is known regarding the physiology of this vasculature. The aim of the present study was to determine the relative responses of equine laminar arteries and veins to the vasoconstrictor agonists phenylephrine (1 nM-10 microM), 5-HT (1 nM-10 microM), PGF2alpha (1 nM-100 microM), and endothelin-1 (1 pM-1 microM). We have determined that laminar veins were more sensitive, with respect to the concentration o...
Chidambaram RM, Eades SC, Moore RM, Hosgood G, Venugopal CS.To characterize the in vitro response of equine cecal longitudinal smooth muscle (CLSM) to endothelin (ET)-1 and assess the role of ETA and ETB receptors in those ET-1-induced responses. Methods: 36 horses without gastrointestinal tract disease. Methods: To determine cumulative concentration-response relationships, CLSM strips were suspended in tissue baths containing graded concentrations of ET-1 (10(-9) to 10(-6)M) with or without BQ-123 (ETA receptor antagonist); with or without IRL-1038 (ETB receptor antagonist); or with both antagonists at concentrations of 10(-9), 10(-7), and 10(-5)M. To...
Mirza MH, Seahorn TL, Oliver JL, Hosgood G, Moore RM.The objective of the study was to determine whether nitric oxide (NO) is present in clinically healthy horses (control) under basal conditions, and if it increases secondary to naturally acquired strangulating large colon volvulus (affected). Eleven affected horses and 10 controls were studied. Jugular venous blood, abdominal fluid, and urine were collected. The NO concentrations were standardized to the creatinine concentration in the respective samples. A biopsy specimen collected from the large colon pelvic flexure at surgery was divided into subsections for processing for inducible nitric ...
Moore RM, Sedrish SA, Holmes EP, Koch CE, Venugopal CS.The objective of this study was to determine and compare the in vitro responses of equine large colon arterial and venous rings to vasodilatory neuropeptides; calcitonin gene-related peptide (CGRP); substance P (SP); vasoactive intestinal polypeptide (VIP); and acetylcholine (ACh), a standard nonpeptide endothelium-dependent vasodilator. Responses of vessel rings to graded concentrations (10(-11) M to 10(-5) M) of each drug were determined in endothelium-intact, denuded, and Nomega-nitro-L-arginine methyl ester (L-NAME, 10(-5) M)-treated rings that were pre-contracted with norepinephrine. Perc...
Wiemer P, Gruys E, van Hoeck B.The aim of this study was to investigate the biological behaviour of vascular grafts replacing a section of the jugular vein in order to improve the results of the surgical treatment of complete thrombosis of the jugular vein in the horse. Seven graft types: fresh allograft, home frozen allograft, glutaraldehyde-fixed allograft, cryo-preserved allograft, PTFE-graft (Gore), small intestinal submucosa preparation (Cook) and fresh autograft, were randomly implanted in ponies. The grafts were removed after one month and examined histologically for: preservation of the graft structures, acceptance ...
Martínez AC, Prieto D, Hernández M, Rivera L, Recio P, García-Sacristán A, Benedito S.This study evaluates the mechanisms underlying endothelium-dependent responses to acetylcholine in horse deep dorsal penile veins. Acetylcholine-induced relaxation was abolished by endothelium removal, the soluble guanylyl cyclase-inhibitor, and the nitric oxide (NO) synthase inhibitors. Acetylcholine-induced relaxation was inhibited by high K+ concentrations and blockade of large-conductance Ca(2+)-activated potassium (BK(Ca)) channels, and voltage-dependent potassium (K(v)) channels. Relaxations were unaffected by a small-conductance K(Ca) (SK(Ca)) channel blocker, or an ATP-sensitive potass...
Kumar P, Timoney JF.The stratified squamous epithelium of the lingual tonsil of five young horses was infiltrated with CD4 and CD8 positive cells, which were very numerous in the crypt reticular epithelium along with macrophages and IgGb and IgA positive cells. Lymphoid follicles of the lamina propria mucosae consisted of a parafollicular area, corona and germinal centre. The parafollicular area was populated by large numbers of CD4 and CD8 positive lymphocytes as well as macrophages, inter-digitating cells, and a few B-lymphocytes. The germinal centre contained mainly IgGb and IgG(T) positive cells, plasma cells...
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...
Prieto D, Arcos LR, Martínez P, Benedito S, García-Sacristán A, Hernández M.The distribution of neuropeptide Y (NPY)-immunorective nerves and the receptors involved in the effects of NPY upon electrical field stimulation (EFS)- and noradrenaline (NA)-elicited contractions were investigated in horse penile small arteries. NPY-immunoreactive nerves were widely distributed in the erectile tissues with a particularly high density around penile intracavernous small arteries. In small arteries isolated from the proximal part of the corpora cavernosa, NPY (30 nM) produced a variable modest enhancement of the contractions elicited by both EFS and NA. At the same concentration...
MacEachern KE, Smith GL, Nolan AM.Hypoxaemia accompanies dorsal recumbency in the horse and frequently complicates general anaesthesia. The physiology associated with this phenomenon is poorly understood. One possible cause of poor tolerance to dorsal recumbency is an absent or reduced response to hypoxic pulmonary vasoconstriction (HPV). This study compared the HPV response in isolated pulmonary artery vessels from equivalent regions of equine and bovine lung. Methods: Equine and bovine, in vitro study. Methods: Equine and bovine pulmonary arteries were removed from the lungs of euthanased horses and cattle. Measurements of i...
Gemensky-Metzler AJ, Wilkie DA, Cook CS.To describe semiconductor diode laser use for anterior uveal cyst deflation and coagulation in dogs, horses and cats. Methods: The presenting clinical signs, surgical technique and postoperative results for four dogs, nine horses and seven cats with anterior uveal cysts treated with diode laser are described. Treated cysts were of sufficient size and/or number to potentially impair vision, damage the corneal endothelium, or increase intraocular pressure (IOP). One dog with free-floating cysts exhibited 'fly biting' behavior. Cysts were suspected of causing shying on the affected side and/or he...
Berhane Y, Bailey SR, Harris PA, Griffiths MJ, Elliott J.Elevated plasma homocysteine (HCy) concentration is a risk factor for cardiovascular diseases associated with endothelial dysfunction, including the human digital ischaemic disease, Raynaud's phenomenon. Objective: HCy causes dysfunction of equine vascular endothelium and elevated plasma concentrations predispose to laminitis. Objective: To determine 1) the concentration of HCy in vitro, which inhibits equine vascular endothelial cell function and 2) any association between risk of laminitis and plasma HCy concentration. Methods: Endothelial function was studied by measuring endothelium-depend...
Ha TY, Kim HS, Shin T.The expression of three isoforms of nitric oxide synthase (NOS) were examined in the testis and epididymis of a thoroughbred horse. Immunohistochemical studies demonstrated the presence of eNOS immunostaining in some germ cells in the seminiferous tubules and in vascular endothelial cells in the interstitial tissues. Interstitial cells, most likely Leydig cells, were also intensely immunopositive for eNOS. The pattern of immunostaining for nNOS was similar to that for eNOS in the testis. Weak expression of iNOS was detected in the seminiferous tubules of the testis, but intense expression was ...
Katz LM, Marr CM, Elliott J.Substance P (SP), a potent vasodilator, has been detected in equine digital sensory-motor nerves. The aim of the study was to characterise the functional responses of equine digital blood vessels to exogenous SP. Pre-constricted equine digital arteries (EDA) and veins (EDV) vasodilated in a biphasic, endothelium- and concentration-dependent manner to SP. A nitric oxide (NO) synthase inhibitor Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME; 300 microm) inhibited both phases of the relaxation response curve of EDAs to SP by >70%. In EDVs, the first relaxant phase to SP was largely...
Katz LM, Marr CM, Elliott J.To compare the responses of equine digital arteries (EDAs) and equine digital veins (EDVs) to endothelin-1 (ET-1) and determine the role of the endothelium and type of receptors involved in the modulation and mediation of those responses, respectively. Methods: 5 to 9 palmar digital vessels/experiment from 28 healthy horses. Methods: Rings of dissected vessels were mounted under tension between force transducer wires in organ baths containing Krebs-Henseleit solution at 30 degrees C. Responses of EDAs and EDVs (with intact [+e] or denuded [-e] endothelium) to cumulative concentrations of ET-1 ...
Herrera HD, Duchene AG, Croxatto JO, Burgués N, Amusquibar H.Vascular tumors of the conjunctiva in the horse are rare. We present a unique case of an intravascular papillary endothelial hyperplasia of the conjunctiva. Methods: Horse. Methods: Case report. A 6-year-old-mare presented with a red mass in the conjunctiva of the left eye. After complete ophthalmologic examination the lesion was excised. The tissue was processed for light microscopy and studied histopathologically. Results: Pathologic examination revealed a nonencapsulated vascular lesion composed of confluent vascular spaces filled by multiple papillary structures composed of a central colla...
Martínez AC, Hernández M, Rivera L, Recio P, García-Sacristán A, Benedito S.To investigate the effect of acetylcholine (ACh) on horse deep dorsal penile vein and to characterize the muscarinic receptor subtypes involved in this response. Methods: Vein rings were mounted in an organ bath chamber, and the isometric tension was recorded. Results: In phenylephrine-contracted veins, ACh (1 nM to 1 microM) induced endothelium-dependent relaxation. The muscarinic receptor antagonist, atropine, produced parallel rightward shifts of the ACh response curves (pA2 = 10.04; pK(B) = 9.98). Carbachol (10 nM to 100 microM) also evoked relaxation in the vein segments, but showed a low...
Elliott J, Bryant CE, Soydan J.Isolated equine digital veins were examined in vitro to study the importance of the endothelium in the responses to both vasodilator and vasoconstrictor agents and to characterise the endothelial-derived mediators involved. Carbachol (Cch; 1 microM) and bradykinin (Bk; 1 nM) caused relaxation of U44069-induced tone by 79.5 +/- 0.35% and 73.7 +/- 4.0% respectively. Mechanical removal of the endothelium completely prevented relaxant responses to Cch and to Bk showing they were mediated by the endothelium. Treatment of veins with NG-nitro-L-arginine methyl ester (L-NAME; 30 and 300 microM) inhibi...
Dubuc V, Lepault E, Theoret CL.Wound repair in horse limbs is often complicated by excessive fibroplasia and scarring. Occlusion of the microvessels populating the granulation tissue appears to be involved in the excessive accumulation of extracellular matrix during the repair of limb wounds. This study aimed to determine whether endothelial cell hypertrophy or hyperplasia, or both, contribute to microvascular occlusion and whether the pericyte is involved in this anomaly. We created 5 wounds, each 2.5 x 2.5 cm, on both forelimbs and on the body of 6 horses. One limb was bandaged to stimulate excessive wound fibroplasia. We...
Wiemer P, Gruys E, van Hoeck B.The aim of this study was to investigate the biological behaviour of vascular grafts replacing a section of the jugular vein in order to improve the results of the surgical treatment of complete thrombosis of the jugular vein in the horse. Seven graft types: fresh allograft, home frozen allograft, glutaraldehyde-fixed allograft, cryo-preserved allograft, PTFE-graft (Gore), small intestinal submucosa preparation (Cook) and fresh autograft, were randomly implanted in ponies. The grafts were removed after one month and examined histologically for: preservation of the graft structures, acceptance ...
McKeever KH, Antas LA, Kearns CF.The objective of the present study was to measure plasma endothelin-1 (ET-1) at rest and during exercise in the horse. Six healthy, Standardbred and Thoroughbred mares (5.3+/-0.8 years; 445.2+/-13.1 kg) which were unfit, but otherwise accustomed to running on the treadmill, were used in the study. Plasma ET-1 concentrations were measured using a commercially available radioimmunoassay kit. Horses performed three trials: a standing control (CON) trial where blood was collected from the jugular vein every minute for 5 min; a graded exercise test (GXT) where blood samples were collected at the en...
Ledbetter EC, Irby NL.To describe the corneal in vivo confocal microscopy (IVCM) findings in horses with putative immune-mediated keratitis (IMMK). Methods: Sixty five horses with IMMK. Methods: Horses diagnosed with IMMK were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module. The findings from the IVCM examinations were correlated with clinical details from ophthalmic examination and diagnostic test results. Results: Eighty eyes from 65 horses were examined. Clinical IMMK lesions were categorized as epithelial (n = 17 eyes), superficial stromal (n = 38), midstromal (n = 18), a...
Benamou AE, Marlin DJ, Callingham BC, Hiley RC, Lekeux R.There is currently little published information about the effects of endothelin-1 (ET-1), a potent endogenous spasmogen of vascular and airway smooth muscle, on pulmonary vasculature and airways or which ET receptor subtypes mediate ET-1-induced vasoconstrictive and bronchoconstrictive action in the horse. Objective: To investigate the effect of endothelin-1 (ET-1) on smooth muscle from isolated equine pulmonary artery and bronchus. In addition, the roles of ETA and ETB receptors in ET-1 mediated contraction in these tissues were assessed. Methods: The force generation of ring segments from pu...
Polikepahad S, Paulsen DB, Moore RM, Costa LR, Venugopal CS.To immunohistochemically determine the expression of endothelin (ET) receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD). Methods: Tissue specimens obtained from 8 healthy and 8 SPAOPD-affected horses. Methods: Horses were examined and assigned to healthy and SPAOPD groups. Horses were then euthanatized, and tissue specimens containing bronchi of approximately 4 to 8 mm in diameter were immediately collected from all lung lobes, fixed in zinc-formalin solution for 12 hours, and embedded in...
Johnson GC, Miller MA, Floss JL, Turk JR.The histologic and immunochemical characteristics of benign vascular tumors excised from the skin of seven young horses were evaluated. The patients were male horses of various breeds and were 16 months of age or younger at the time of presentation. Six tumors occurred on the extremities, and one was removed from the lip. Histologically, most tumors consisted of cellular nodules of varying compactness with few to many blood-filled lumina. Nodular growth separated preexisting adnexa and subcutaneous collagen. Some tumors contained or consisted predominantly of loosely packed arrays of ramifying...
Söder J, Bröjer JT, Nostell KE.Systemic hypertension is a prominent feature in humans with metabolic syndrome (MS) and this is partly caused by an enhanced endothelin-1 (ET-1) mediated vasoconstriction. There are indications that systemic hypertension might be a feature in equine metabolic syndrome (EMS) but if ET-1 is involved in the development of hypertension in horses is not known. Increased levels of cortisol have also been found in humans with MS but there are no reports of this in horses. Before blood pressure, plasma ET-1 and serum cortisol can be evaluated in horses with EMS, it is necessary to investigate the inte...
Wunn D, Wardrop KJ, Meyers K, Kramer J, Ragle C.To develop methods to isolate, culture, and characterize equine hoof endothelial cells (EC) and keratinocytes. Methods: Cells harvested from the forelimbs of 8 horses. Methods: EC were obtained via catheters placed in the palmar digital arteries of the disarticulated lower portion of the forelimbs from 4 horses that had been heparinized prior to euthanasia. Phosphate-buffered saline solution was used to remove and discard RBC from blood vessels, and collagenase was used to loosen and flush EC from the vasculature. Hoof keratinocytes were obtained from 4 recently euthanatized horses by use of d...
Sharpe AN, Seeto WJ, Winter RL, Zhong Q, Lipke EA, Wooldridge AA.OBJECTIVE To evaluate optimal isolation of endothelial colony-forming cells (ECFCs) from peripheral blood of horses. SAMPLE Jugular and cephalic venous blood samples from 17 adult horses. PROCEDURES Each blood sample was divided; isolation was performed with whole blood adherence (WBA) and density gradient centrifugation (DGC). Isolated cells were characterized by uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled acetylated low-density lipoprotein (DiI-Ac-LDL), vascular tubule formation, and expression of endothelial (CD34, CD105, vascular endothelial growth ...
Robertson TP, Moore JN, Noschka E, Lewis TH, Lewis SJ, Peroni JF.To determine the effects of inhibition of Rho-kinase or Src-family protein tyrosine kinases (srcPTK) on agonist-induced contractile responses in equine laminar arteries and veins. Methods: Laminar arteries and veins obtained from 13 adult mixed-breed horses. Methods: Laminar vessels were mounted on myographs and exposed to phenylephrine (PE), 5-hydroxytryptamine (5-HT), prostaglandin F(2) (PGF(2)), and endothelin-1 (ET-1) with or without the Rho-kinase inhibitor Y-27632 (10 microM), srcPTK inhibitor PP2 (10 microM), or a negative control analogue for PP2 (PP3; 10 microM). Results: Responses to...
Prieto D, Arcos LR, Martínez P, Benedito S, García-Sacristán A, Hernández M.The distribution of neuropeptide Y (NPY)-immunorective nerves and the receptors involved in the effects of NPY upon electrical field stimulation (EFS)- and noradrenaline (NA)-elicited contractions were investigated in horse penile small arteries. NPY-immunoreactive nerves were widely distributed in the erectile tissues with a particularly high density around penile intracavernous small arteries. In small arteries isolated from the proximal part of the corpora cavernosa, NPY (30 nM) produced a variable modest enhancement of the contractions elicited by both EFS and NA. At the same concentration...
Katz LM, Marr CM, Elliott J.To compare the responses of equine digital arteries (EDAs) and equine digital veins (EDVs) to endothelin-1 (ET-1) and determine the role of the endothelium and type of receptors involved in the modulation and mediation of those responses, respectively. Methods: 5 to 9 palmar digital vessels/experiment from 28 healthy horses. Methods: Rings of dissected vessels were mounted under tension between force transducer wires in organ baths containing Krebs-Henseleit solution at 30 degrees C. Responses of EDAs and EDVs (with intact [+e] or denuded [-e] endothelium) to cumulative concentrations of ET-1 ...
Cogswell AM, Johnson PJ, Adams HR.To test the hypothesis that endothelium-derived nitric oxide modulates vasomotor reactivity in equine digital arteries. Methods: Digital arteries were isolated from adult horses, and their vasodilator properties were examined in an in vitro controlled environment. Methods: Five adult horses (1 gelding, 4 mares) without evidence of hoof or vascular disease were studied. Methods: Arterial rings with or without endothelium were exposed to endothelium-dependent vasodilator drugs in the presence or absence of a pharmacologic inhibitor of the enzyme nitric oxide synthase. Results: Vasodilator effect...
Katz LM, Marr CM, Elliott J.Substance P (SP), a potent vasodilator, has been detected in equine digital sensory-motor nerves. The aim of the study was to characterise the functional responses of equine digital blood vessels to exogenous SP. Pre-constricted equine digital arteries (EDA) and veins (EDV) vasodilated in a biphasic, endothelium- and concentration-dependent manner to SP. A nitric oxide (NO) synthase inhibitor Nomega-nitro-L-arginine methyl ester hydrochloride (L-NAME; 300 microm) inhibited both phases of the relaxation response curve of EDAs to SP by >70%. In EDVs, the first relaxant phase to SP was largely...
Eades SC, Stokes AM, Johnson PJ, LeBlanc CJ, Ganjam VK, Buff PR, Moore RM.To quantify changes in endothelium-derived factors and relate those changes to various aspects of digital hemodynamics during the prodromal stages of carbohydrate overload (CHO)-induced laminitis in horses. Methods: 20 adult horses without abnormalities of the digit. Methods: Digital and jugular venous blood samples were collected at 1-hour intervals (for assessment of endothelin-1 [ET-1] immunoreactivity and measurement of glucose, insulin, and nitric oxide [NO] concentrations) or 4-hour intervals (CBC and platelet-neutrophil aggregate assessment) for 8 hours or 16 hours after induction of CH...
Goehring LS, Brandes K, Ashton LV, Wittenburg LA, Olea-Popelka FJ, Lunn DP, Soboll Hussey G.Equine herpesvirus-associated myeloencephalopathy is the result of endothelial cell infection of the spinal cord vasculature with equine herpesvirus-1 (EHV-1) during cell-associated viraemia. Endothelial cell infection requires contact between infected peripheral blood mononuclear and endothelial cells. Inflammation generated during viraemia likely upregulates adhesion molecule expression on both cell types increasing contact and facilitating endothelial cell infection. Objective: Evaluating the role of anti-inflammatory drugs in decreasing endothelial cell infection with EHV-1. Methods: In vi...
Junginger J, Rötting A, Staszyk C, Kramer K, Hewicker-Trautwein M.An adult horse was presented with a mass located within the dermis and subcutis of the right forelimb. At post-mortem examination there was a marked increase in thickness of the subcutis of the right forelimb extending from the fetlock to the pectoral muscles. Microscopically, the dermis and subcutis were diffusely infiltrated by endothelial-like cells forming irregular channels and cavernous spaces. Immunohistochemical examination revealed the endothelial-like cells to express vimentin, factor VIII-related antigen and PROX-1, confirming the identity of the tumour as a lymphangioma. Ultrastruc...
Lamar CH, Turek JJ, Bottoms GD, Fessler JF.Certain in vitro culture conditions were determined for equine endothelial cells obtained from the aorta and pulmonary arteries. Cells were enzymatically isolated from the vessel lumen, using clostridial collagenase (2.5 mg/ml of Hanks's balanced salt solution) incubated at 37 C for 30 minutes. Cells were cultured in alpha minimum essential medium supplemented with plasma-derived and nonplasma-derived bovine fetal sera, endothelial cell-growth supplement, heparin, and antibiotics. Smooth muscle cell growth was not inhibited with nonplasma-derived animal sera, plasma-derived equine serum, or he...
Wille KH, Schnorr B.Arterioles, precapillary sphincters, capillary endothelium, and pericytes probably regulate the blood flow in the intestinal microvascular bed similar to other regions of the body because of their equipment with contractile filaments. Only throttle veins with their arrangement of pools and their characteristics probably exert influence on the hemodynamic qualities of the blood flow in the intestinal mucosa.
Menzies-Gow NJ, Wray H, Bailey SR, Harris PA, Elliott J.Insulin and inflammatory cytokines may be involved in equine laminitis, which might be associated with digital vascular dysfunction. This study determined the effects of TNF-α and insulin on the endothelial-dependent relaxant responses of equine digital blood vessels and on equine digital vein endothelial cell (EDVEC) cGMP production. Methods: Isolated rings of equine digital arteries (EDAs) and veins (EDVs) were obtained and EDVECs were cultured from horses euthanized at an abattoir. Methods: The effect of incubation with TNF-α (10 ng/ml) and/or insulin (1,000 μIU/ml) for 1.5 h or overn...
Stokes AM, Venugopal CS, Hosgood G, Eades SC, Moore RM.The goals of this study were to determine the concentration-response (C-R) relationship of endothelin-1 (ET-1), compare 2 ET-receptor antagonists and determine the antagonist concentrations that block the vasomotor effects of ET-1, and compare the effectiveness of ET-1 and previously studied vasoconstrictors in equine palmar digital arterial and venous rings in vitro. Vessel rings from 8 nonlaminitic horses were placed in Tyrode's solution, 1 side fixed to the floor of an organ bath and the other side fixed to a force-displacement transducer. Two separate studies were conducted: (I) incubation...
Martínez AC, Prieto D, Hernández M, Rivera L, Recio P, García-Sacristán A, Benedito S.This study evaluates the mechanisms underlying endothelium-dependent responses to acetylcholine in horse deep dorsal penile veins. Acetylcholine-induced relaxation was abolished by endothelium removal, the soluble guanylyl cyclase-inhibitor, and the nitric oxide (NO) synthase inhibitors. Acetylcholine-induced relaxation was inhibited by high K+ concentrations and blockade of large-conductance Ca(2+)-activated potassium (BK(Ca)) channels, and voltage-dependent potassium (K(v)) channels. Relaxations were unaffected by a small-conductance K(Ca) (SK(Ca)) channel blocker, or an ATP-sensitive potass...
Vairo S, Favoreel H, Scagliarini A, Nauwynck H.Currently, little is known on the cellular pathogenesis of equine arteritis virus (EAV). The purpose of the present study was to identify the target cells in ponies experimentally inoculated with EAV 08P178 (EU, clade-1). EAV-target organs (respiratory tissues with associated lymphoid tissues and large intestines), collected at 3 and 7 days post inoculation (dpi) and with virus titers≥10(5.0) TCID50/g, were processed with double immunofluorescence staining for the simultaneous detection of EAV N-protein and one of the following cell markers: CD172a (myeloid cells), CD3 (T lymphocytes), IgM (...
Kumar P, Timoney JF.The stratified squamous epithelium of the lingual tonsil of five young horses was infiltrated with CD4 and CD8 positive cells, which were very numerous in the crypt reticular epithelium along with macrophages and IgGb and IgA positive cells. Lymphoid follicles of the lamina propria mucosae consisted of a parafollicular area, corona and germinal centre. The parafollicular area was populated by large numbers of CD4 and CD8 positive lymphocytes as well as macrophages, inter-digitating cells, and a few B-lymphocytes. The germinal centre contained mainly IgGb and IgG(T) positive cells, plasma cells...
Elliott J, Brady FE.Isolated equine digital veins (EDVs) which had been denuded of their endothelium were used to study adenosine receptors causing vasodilation. When the blood vessel wall tension was raised with the thromboxanemimetic, U44069 (30 nM), the order of vasodilator potency of adenosine receptor agonists was: 5'-N-ethylcarboxamidoadenosine (NECA) > 2-p-(2-carboxyethyl)phenyl amino-5'-N-ethylcarboxamido-adenosine (CGS 21680) > 5'-N-methylcarboxamido-adenosine (MECA) > N6-cyclohexyladenosine (CHA) > N6-cyclopentyladenosine (CPA) > N6-2-(4-Aminophenyl)ethyladenosine (APNEA) > adenosine. ...
Finding EJT, Lawson C, Elliott J, Harris PA, Menzies-Gow NJ.Microvesicles are small (up to 1 μm) vesicles found in plasma and other bodily fluids. They are recognised as part of the normal system of inter-cellular communication but altered numbers are also used as biomarkers of disease. Microvesicles have not been studied in detail in the horse but may be relevant to diseases such as laminitis. Identification of equine cell specific microvesicles was performed by developing a panel of cross reactive antibodies to use in flow cytometry to detect microvesicles of platelet, leucocyte and endothelial origin in plasma from healthy ponies and those predis...
Gauff FC, Patan-Zugaj B, Licka TF.To determine the effect of short-term hyperinsulinemia on the localization and expression of endothelin receptor (ETR)-A and ETR-B in lamellar tissue of the forelimbs of horses. Methods: Distal portion of 15 cadaveric forelimbs from healthy adult horses (1 limb/horse) obtained immediately after slaughter at an abattoir. Methods: Each forelimb was assigned to 1 of 3 treatment groups (perfused with autologous blood for 10 hours [control perfusion; n = 5], perfused with an insulin [142 ± 81 μU/mL] perfusate for 10 hours [insulinemic perfusion; 5], or not perfused [unperfused control; 5]). Immun...
Miyamoto A, Obi T, Nishio A.The vasomotor effects of 5-hydroxytryptamine (5-HT) on isolated equine basilar arteries were studied. 5-HT induced contractions of equine basilar arteries in a concentration-dependent manner, with a pEC50 value (with 95% confidence limits) of 7.35 (7.08-7.62). Similar results were obtained with endothelium-denuded basilar arteries. Contractions were not competitively inhibited by the 5-HT2 receptor antagonist ketanserin at low concentrations of 5-HT. Conversely, at high concentrations of 5-HT, contractions were inhibited by ketanserin in a concentration-dependent manner, with a pA2 value of 8....
