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Topic:Oxidative Stress

Oxidative stress in horses refers to an imbalance between the production of reactive oxygen species (ROS) and the ability of the horse's body to detoxify these reactive intermediates or repair the resulting damage. It can occur as a result of various physiological and environmental factors, such as exercise, illness, or exposure to toxins. Oxidative stress can lead to cellular damage, affecting lipids, proteins, and DNA. In horses, it is studied in the context of its potential impact on performance, health, and disease progression. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, effects, and management of oxidative stress in equine health.
Activation of equine neutrophils by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine induces a different response in reactive oxygen species production and release of active myeloperoxidase.
Veterinary immunology and immunopathology    March 4, 2009   Volume 130, Issue 3-4 243-250 doi: 10.1016/j.vetimm.2009.02.015
Franck T, Kohnen S, de la Rebière G, Deby-Dupont G, Deby C, Niesten A, Serteyn D.Neutrophil (PMN) contribution to the acute inflammatory processes may lead to an excessive generation of reactive oxygen metabolites species (ROS) and secretion of granule enzymes. We compared the effects of either phorbol myristate acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP) in combination with a pre-treatment by cytochalasin B (CB) on the production of ROS and the release of total and active myeloperoxidase (MPO) by isolated equine PMNs. The ROS production was assessed by lucigenin dependent chemiluminescence (CL) and ethylene release by alpha-keto-gamma-methylthiobutyric...
The role of dietary antioxidant insufficiency on the permeability of the blood-brain barrier.
Journal of neuropathology and experimental neurology    November 20, 2008   Volume 67, Issue 12 1187-1193 doi: 10.1097/NEN.0b013e31818f8f51
Mohammed HO, Starkey SR, Stipetic K, Divers TJ, Summers BA, de Lahunta A.Our previous studies implicated vitamin E deficiency as a risk factor for equine motor neuron disease, a possible model of human amyotrophic lateral sclerosis, and showed direct effects of this deficiency on brain vascular endothelium. To gain better understanding of the pathogenesis of equine motor neuron disease, we determined the effects of dietary antioxidant insufficiency and the resultant brain tissue oxidative stress on blood-brain barrier permeability. Rats (n = 40) were maintained on a diet deficient of vitamin E for 36 to 43 weeks; 40 controls were fed a normal diet. Permeability of ...
The stability of the reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) tests on stored horse blood.
Veterinary journal (London, England : 1997)    November 17, 2008   Volume 183, Issue 2 217-218 doi: 10.1016/j.tvjl.2008.09.018
Celi P, Sullivan M, Evans D.Increasing interest in the role of oxidative stress (OS) in equine medicine has highlighted the need to develop reliable methods to quantify it. In this study we describe the effect of refrigeration (at 4 degrees C) on the stability of the reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP) tests carried out on 15 healthy horses. Blood samples, collected from the jugular vein, were immediately placed on ice and analysed using both the d-ROMs and BAP tests. Samples were also refrigerated at 4 degrees C and tested after 3, 7 and 24 h. The average results were similar ...
Tissue concentrations of 4-HNE in the black walnut extract model of laminitis: indication of oxidant stress in affected laminae.
Veterinary immunology and immunopathology    November 11, 2008   Volume 129, Issue 3-4 211-215 doi: 10.1016/j.vetimm.2008.11.016
Yin C, Pettigrew A, Loftus JP, Black SJ, Belknap JK.In the septic horse prone to laminitis, a similar activation of the innate immune system appears to occur as reported in the septic human prone to organ failure. Because oxidant injury plays a central role in organ failure occurring due to an overzealous innate immune response in human sepsis, this study was performed to determine whether there was evidence of oxidant stress in the laminar tissue in the early stages of laminitis. 4-Hydroxy-2-nonenal (4-HNE), a lipid aldehyde that forms due to lipid peroxidation occurring during episodes of oxidant stress, readily forms adducts with cellular pr...
The effect of methyl sulphonyl methane supplementation on biomarkers of oxidative stress in sport horses following jumping exercise.
Acta veterinaria Scandinavica    November 7, 2008   Volume 50, Issue 1 45 doi: 10.1186/1751-0147-50-45
Marañón G, Muñoz-Escassi B, Manley W, García C, Cayado P, de la Muela MS, Olábarri B, León R, Vara E.Exercise induces changes in several organs and tissues, and this process might be due to oxidative damage caused by free radicals and inflammatory mediators. Methyl Sulphonyl Methane, better known as MSM, is a naturally occurring sulphur compound with well-known antioxidant properties. On the other hand, Vitamin C is important in limiting free radical damage in the aqueous phase of the cell, and cellular vitamin C status may be linked to the mechanisms involved in quenching cellular reactive oxygen species. The aim of this study was to determine if supplementation with MSM and vitamin C could ...
Inflammatory and redox status of ponies with a history of pasture-associated laminitis.
Veterinary immunology and immunopathology    November 7, 2008   Volume 129, Issue 3-4 216-220 doi: 10.1016/j.vetimm.2008.11.004
Treiber K, Carter R, Gay L, Williams C, Geor R.Inflammatory and redox signals could render lamellar tissue susceptible to damage and contribute to higher risk for laminitis in obese or insulin resistant ponies just as these factors contribute to health risks in humans with metabolic syndrome. This study evaluated circulating markers of inflammatory and redox status in ponies that had a history of recurrent bouts of pasture-associated laminitis (PL, n = 42) or had never developed clinical laminitis (NL, n = 34) under the current management conditions. There were no differences (P > 0.05) between PL and NL ponies for markers of antioxidan...
Alpha-tocopherol protects against oxidative damage to lipids of the rod outer segments of the equine retina.
Veterinary journal (London, England : 1997)    October 1, 2008   Volume 182, Issue 3 463-468 doi: 10.1016/j.tvjl.2008.08.008
Terrasa AM, Guajardo MH, Marra CA, Zapata G.Oxidative stress is a possible risk factor for eye diseases. Lipid peroxidation is one of the major events induced by oxidative stress and is particularly active in polyunsaturated fatty acid (PUFA)-rich biomembranes. This work evaluated endogenous lipid antioxidants, in vitro non-enzymatic lipid peroxidation of rod outer segment membranes (ROS), the fatty acid composition during oxidative damage of total lipids from equine retina and ROS, and the protective action of alpha-tocopherol (alpha-Toc). The major lipid soluble antioxidant was alpha-Toc followed by retinoids and carotenoids. The reti...
Oxidative stress, osmotic stress and apoptosis: impacts on sperm function and preservation in the horse.
Animal reproduction science    April 29, 2008   Volume 107, Issue 3-4 257-267 doi: 10.1016/j.anireprosci.2008.04.014
Ball BA.Oxidative stress is an important component of the cytopathology of equine spermatozoa undergoing storage as liquid or frozen semen. Damage to chromatin, membranes and proteins of sperm are important components of oxidative damage to sperm. Similarly, sperm are exposed to a variety of osmotic stresses during storage that result from exposure to hypertonic media or result as a consequence of osmotic changes induced during freezing. A number of changes induced during processing and storage of equine sperm also appear to induce apoptotic-like changes which may adversely affect sperm survival and f...
Oxidative stress or oxidant/antioxidant dis-equilibrium in horses? A hot topic.
Veterinary journal (London, England : 1997)    March 7, 2008   Volume 177, Issue 2 157-158 doi: 10.1016/j.tvjl.2007.11.005
Williams CA.No abstract available
Free radicals and antioxidants in inflammatory processes and ischemia-reperfusion injury.
The Veterinary clinics of North America. Small animal practice    February 6, 2008   Volume 38, Issue 1 31-v doi: 10.1016/j.cvsm.2007.11.008
Vajdovich P.This article discusses the current understanding of the role of free radicals and antioxidants in inflammatory processes and in ischemia reperfusion injury. It begins by describing the manifestations of acute inflammation and outlining the cellular events that occur during inflammation. It then describes the biochemical mediators of inflammation with special attention to nitric oxide. It details the process of hypoxia reperfusion injury, the enzymes involved, its treatment, and studies involving specific hypoxia reperfusion injuries in various animal species.
Apoptosis and antioxidant status are influenced by age and exercise training in horses.
Journal of animal science    December 21, 2007   Volume 86, Issue 3 576-583 doi: 10.2527/jas.2007-0585
Williams CA, Gordon ME, Betros CL, McKeever KH.Eight mature (12 +/- 2 yr; MAT) and 5 older (22 +/- 2 yr; OLD) Standardbred mares were used to test the hypothesis that aging and exercise training would alter apoptosis in white blood cells and antioxidant status. The horses were housed indoors overnight (16 h/d) in 3 m x 3 m stalls and were turned out in a drylot during the day. They were fed a diet consisting of total mixed ration, hay cubes fed ad libitum or an equine senior diet plus grass hay. Horses were trained for 20 to 30 min/d, 3 to 5 d/wk for 8 wk at a submaximal work intensity between 60 to 70% of maximal heart rate. A graded exer...
Change in blood antioxidant status of horses moved from a stable following diagnosis of equine motor neuron disease.
The Canadian veterinary journal = La revue veterinaire canadienne    December 7, 2007   Volume 48, Issue 11 1165-1167 
Delguste C, de Moffarts B, Kirschvink N, Art T, Pincemail J, Defraigne JO, Amory H, Lekeux P.The antioxidant status of 10 horses living in stable 1 where 2 cases of equine motor neuron disease had previously been diagnosed was assessed before and 9 weeks after moving to another stable. Duration of residence in stable 1, subsequent moving, or both, significantly affected several parameters of the antioxidant status. Le statut antioxydatif de 10 chevaux logés dans l’écurie 1, où 2 cas de maladie du neurone moteur avaient été précédemment diagnostiqués, a été vérifié avant et 9 semaines après leur transfert dans une autre écurie. La durée de l’hébergement dans l’é...
Effect of exercise on development of capillary supply and oxidative capacity in skeletal muscle of horses.
American journal of veterinary research    November 3, 2007   Volume 68, Issue 11 1226-1231 doi: 10.2460/ajvr.68.11.1226
Rietbroek NJ, Dingboom EG, Schuurman SO, Hengeveld-van der Wiel E, Eizema K, Everts ME.To determine whether postnatal development of oxidative capacity and capillary supply of skeletal muscle is affected by various movement regimens in horses. Methods: 35 foals. Methods: Dutch Warmblood foals were allocated into 3 groups (box stall rest, box stall rest with training, and free pasture exercise). Training comprised an increasing number of gallop sprints from 1 week after birth to 22 weeks of age. From 22 to 48 weeks, the 3 groups were combined and allowed to exercise freely. Capillary supply (diffusion index [ie, area supplied by 1 capillary]), citrate synthase (CS) activity, and ...
Direct stimulation of the oxidative activity of isolated equine neutrophils by TNF-alpha and IL-1beta.
Veterinary immunology and immunopathology    October 2, 2007   Volume 121, Issue 1-2 101-106 doi: 10.1016/j.vetimm.2007.09.006
Benbarek H, Deby-Dupont G, Deby C, Serteyn D.The capacity of the two cytokines TNF-alpha and IL-1beta to directly stimulate the oxidative activity of polymorphonuclear neutrophils remains debated. The purpose of this study was to verify if a direct stimulation of equine neutrophils by TNF-alpha and IL-1beta was possible. Equine neutrophils were isolated from blood by discontinuous density gradient centrifugation. The cell viability after isolation was >98%. The neutrophils were used at 1.25 x 10(6) cells by assay, immediately after isolation. The oxidative activity of neutrophils was measured by luminol- or lucigenin-enhanced chemilumine...
The oxidant/antioxidant equilibrium in horses.
Veterinary journal (London, England : 1997)    September 25, 2007   Volume 177, Issue 2 178-191 doi: 10.1016/j.tvjl.2007.07.033
Kirschvink N, de Moffarts B, Lekeux P.Since "free radical research" started in 1954, understanding the role of oxidants and antioxidants in physiological and pathological conditions has increased continuously. Oxidants are essentially generated by metabolic enzymes, inflammatory cells and mitochondrial electron leakage; they are indispensable for the cellular redox regulation and may, under certain conditions, have a pro-inflammatory stimulatory role. Endogenous and exogenous antioxidants counterbalance the oxidative processes and so maintain the oxidant/antioxidant equilibrium. Excessive oxidant generation or antioxidant insuffic...
Serum opsonization capacity, phagocytosis, and oxidative burst activity in neonatal foals in the intensive care unit.
Journal of veterinary internal medicine    August 22, 2007   Volume 21, Issue 4 797-805 doi: 10.1892/0891-6640(2007)21[797:socpao]2.0.co;2
Gardner RB, Nydam DV, Luna JA, Bicalho ML, Matychak MB, Flaminio MJ.Phagocytic activity of neonatal foals has been reported to be similar to that of adult horses, but serum opsonization capacity develops with age and may be further altered when opsonins are consumed during infection. Objective: Phagocytosis, oxidative burst activity, and serum opsonization capacity in neonatal foals admitted to an intensive care unit are reduced in comparison with control foals. Methods: Blood samples were collected from hospitalized neonatal foals and from control foals. Hospitalized foals were characterized as sick or septic on the basis of a sepsis score and received intrav...
Horse heart myoglobin catalyzes the H2O2-dependent oxidative dehalogenation of chlorophenols to DNA-binding radicals and quinones.
Biochemistry    August 4, 2007   Volume 46, Issue 34 9823-9829 doi: 10.1021/bi700684u
Osborne RL, Coggins MK, Walla M, Dawson JH.The heme-containing respiratory protein, myoglobin (Mb), best known for oxygen storage, can exhibit peroxidase-like activity under conditions of oxidative stress. Under such circumstances, the initially formed ferric state can react with H2O2 (or other peroxides) to generate a long-lived ferryl [Fe(IV)=O] Compound II (Cpd II) heme intermediate that is capable of oxidizing a variety of biomolecules. In this study, the ability of Mb Cpd II to catalyze the oxidation of carcinogenic halophenols is demonstrated. Specifically, 2,4,6-trichlorophenol (TCP) is converted to 2,6-dichloro-1,4-benzoquinone...
Inhibitory effect of curcuminoids and tetrahydrocurcuminoids on equine activated neutrophils and myeloperoxidase activity.
Physiological research    July 26, 2007   Volume 57, Issue 4 577-587 doi: 10.33549/physiolres.931086
Franck T, Kohnen S, Grulke S, Neven P, Goutman Y, Peters F, Pirotte B, Deby-Dupont G, Serteyn D.In the horse, the inflammation response to various pathologies (intestinal strangulations, laminitis, etc.) involves an excessive stimulation of the polymorphonuclear neutrophils releasing reactive oxygen species (ROS) and myeloperoxidase (MPO). The aim of the present work was to study the effect of natural polyphenols, curcuminoids and tetrahydrocurcuminoids (THC) on isolated stimulated equine neutrophils and on the activity of purified MPO. The ROS production and the release of MPO by activated neutrophils were measured by chemiluminescence and ELISA techniques, respectively. The activity of...
Oral vitamin E supplementation on oxidative stress, vitamin and antioxidant status in intensely exercised horses.
Equine veterinary journal. Supplement    April 4, 2007   Issue 36 617-621 doi: 10.1111/j.2042-3306.2006.tb05614.x
Williams CA, Carlucci SA.Vitamin E is the most commonly supplemented antioxidant in horses; however, previous research is not conclusive as to the recommended level for exercising horses. Objective: To evaluate the effects of 3 levels of vitamin E supplementation on oxidative stress and vitamin/antioxidant status in intensely exercised horses to determine the optimal level of vitamin E supplementation. Methods: Twelve unfit Standardbreds were divided into 3 groups, supplemented orally with 0 (CON), 5000 (MOD), or 10,000 (HI) iu/day of DL-alpha-tocopheryl acetate. The 3 x 3 Latin square design consisted of three 4 week...
Plasma concentrations of myeloperoxidase in endurance and 3-day event horses after a competition.
Equine veterinary journal. Supplement    April 4, 2007   Issue 36 298-302 doi: 10.1111/j.2042-3306.2006.tb05557.x
Art T, Franck T, Gangl M, Votion D, Kohnen S, Deby-Dupont G, Serteyn D.In man, exercise of any type has been shown to induce neutrophil degranulation and respiratory burst activity, as well as an increase in plasma myeloperoxidase (MPO), a specific enzyme of neutrophil azurophilic granules with a strong oxidative activity. Until now, it is not known whether this is the same in horses. Objective: To study whether degranulation of blood neutrophils may be induced by exercise by measuring the total concentration of MPO and assess the possible influence of type of competition on this exercise-induced adjustment. Methods: Blood was sampled before, and 30 min after, th...
Investigation of blood oxidant/antioxidant markers in healthy competition horses of different breeds.
Equine veterinary journal. Supplement    April 4, 2007   Issue 36 239-244 doi: 10.1111/j.2042-3306.2006.tb05546.x
Kirschvink N, de Moffarts B, Farnir F, Pincemail J, Lekeux P.There is increasing evidence that the equine athlete is exposed to exercise-induced changes of its oxidant/antioxidant balance and antioxidant supplementation is frequently recommended. However, it is unknown whether there is a specific need for antioxidants according to performance, breed, gender or age. Objective: To assess whether breed-, gender- and age-related differences of blood oxidant/antioxidant markers occur in competition horses. Methods: Healthy horses (n = 493) underwent oxidant/ antioxidant blood marker determination. Vitamin E, lipophilic antioxidant capacity (ACL), ascorbic ac...
The quantification of lipid and protein oxidation in stallion spermatozoa and seminal plasma: seasonal distinctions and correlations with DNA strand breaks, classical seminal parameters and stallion fertility.
Animal reproduction science    March 30, 2007   Volume 106, Issue 1-2 36-47 doi: 10.1016/j.anireprosci.2007.03.020
Morte MI, Rodrigues AM, Soares D, Rodrigues AS, Gamboa S, Ramalho-Santos J.The goal of this work was to correlate oxidative stress caused by reactive oxygen species (ROS) and DNA damage with classic semen parameters in spermatozoa and seminal plasma of fertile and subfertile stallions. Oxidation was measured in both lipids and proteins, using the thiobarbituric acid reactive species (TBARS) assay and the DNPH carbonyl groups assay, respectively. Sperm DNA damage was monitored using the TUNEL assay. These parameters were monitored in samples obtained during the breeding and the non-breeding seasons. In general, fertile stallions showed better classical semen parameter...
Trauma with neurologic sequelae.
The Veterinary clinics of North America. Equine practice    March 24, 2007   Volume 23, Issue 1 81-101 doi: 10.1016/j.cveq.2006.11.003
Tennent-Brown BS.Spinal cord injury (SCI) in horses may arise from rearing and falling backward, collisions, kicks, and slipping. The pathophysiology of SCI comprises a primary mechanical injury followed by a cascade of secondary events. These secondary events include microvascular ischemia, oxidative stress, excitotoxicity, ion dysregulation, and inflammation. It is often the severity of secondary injury that limits the restoration of neurologic function. Clinical signs after SCI depend on the location of the lesion and the relative amount of damage to the gray and white matter. Acute management of SCI should...
Oxidative stress.
The Veterinary clinics of North America. Equine practice    March 24, 2007   Volume 23, Issue 1 135-157 doi: 10.1016/j.cveq.2006.11.004
Soffler C.Oxidative stress refers to the cellular injury and pathologic change that occurs when there is an imbalance favoring oxidants over antioxidants within a living organism. In human medicine, oxidative stress has been implicated in numerous disease processes, which has led to further research into the clinical benefits and efficacy of antioxidant therapy. The evaluation of oxidative stress in the horse has been limited primarily to ischemia-reperfusion injury of the gastrointestinal tract, recurrent airway obstruction, exercise, osteoarthritis, equine motor neuron disease, and pituitary pars inte...
Protective effect of magnesium and potassium ions on the permeability of the external mitochondrial membrane.
Archives of biochemistry and biophysics    January 29, 2007   Volume 461, Issue 1 13-23 doi: 10.1016/j.abb.2007.01.007
Gorgoglione V, Laraspata D, La Piana G, Marzulli D, Lofrumento NE.The data reported are fully consistent with the well-known observation that exogenous cytochrome c (cyto-c) molecules do not permeate through the outer membrane of mitochondria (MOM) incubated in isotonic medium (250 mM sucrose). Cyto-c is unable to accept electrons from the sulfite/cyto-c oxido-reductase (Sox) present in the intermembrane space, unless mitochondria are solubilized. Mitochondria incubated in a very high hypotonic medium (25 mM sucrose), in contrast to any expectation, continue to be not permeable to added cyto-c even if Sox and adenylate kinase are released into the medium. Th...
Effects of intensity and duration of exercise on muscular responses to training of thoroughbred racehorses.
Journal of applied physiology (Bethesda, Md. : 1985)    January 25, 2007   Volume 102, Issue 5 1871-1882 doi: 10.1152/japplphysiol.01093.2006
Rivero JL, Ruz A, Martí-Korff S, Estepa JC, Aguilera-Tejero E, Werkman J, Sobotta M, Lindner A.This study examined the effects of the intensity and duration of exercise on the nature and magnitude of training adaptations in muscle of adolescent (2-3 yr old) racehorses. Six thoroughbreds that had been pretrained for 2 mo performed six consecutive conditioning programs of varying lactate-guided intensities [velocities eliciting blood lactate concentrations of 2.5 mmol/l (v2.5) and 4 mmol/l (v4), respectively] and durations (5, 15, 25 min). Pre- and posttraining gluteus muscle biopsies were analyzed for myosin heavy chain content, fiber-type composition, fiber size, capillarization, and fi...
Laminar xanthine oxidase, superoxide dismutase and catalase activities in the prodromal stage of black-walnut induced equine laminitis.
Equine veterinary journal    January 19, 2007   Volume 39, Issue 1 48-53 doi: 10.2746/042516406x151320
Loftus JP, Belknap JK, Stankiewicz KM, Black SJ.REASONS FOR STUDY: Xanthine oxidase (XO)-dependent production of superoxide anion and hydrogen peroxide, a characteristic of ischaemia-reperfusion injury, may contribute to the development of equine laminitis. Objective: To determine the levels of XO and antioxidant enzymes (catalase, superoxide dismutase [SOD]) in the digital laminae of normal horses (CON) and horses in the developmental stage of laminitis using the black walnut extract (BWE) model. Methods: Healthy horses (n = 12) were administered BWE (BWE group, n = 6), or water (CON group, n = 6) through a nasogastric tube. At the onset o...
Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses.
Veterinary journal (London, England : 1997)    November 15, 2006   Volume 174, Issue 1 113-121 doi: 10.1016/j.tvjl.2006.06.001
De Moffarts B, Portier K, Kirschvink N, Coudert J, Fellmann N, van Erck E, Letellier C, Motta C, Pincemail J, Art T, Lekeux P.The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water an...
Anti-oxidative status and semen quality during cooled storage in stallions.
Journal of veterinary medicine. A, Physiology, pathology, clinical medicine    October 24, 2006   Volume 53, Issue 9 486-489 doi: 10.1111/j.1439-0442.2006.00879.x
Pagl R, Aurich C, Kankofer M.Activity of the anti-oxidative enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), content of thiobarbituric acid reactive substances (TBARS) and SH-groups were determined in native stallion semen (n = 8 stallions). Semen was then diluted in Kenney extender, EquiPro((R)) extender either with or without addition of N-acetyl cysteine or phosphate-buffered saline (PBS) and stored for 72 h at 5 degrees C. Correlations between initial activity of enzymes and development of semen motility and membrane integrity were calculated. Activities of GSH-Px, SOD and CAT im...
Generation of superoxide anion by equine spermatozoa as detected by dihydroethidium.
Theriogenology    October 12, 2006   Volume 67, Issue 3 580-589 doi: 10.1016/j.theriogenology.2006.07.021
Burnaugh L, Sabeur K, Ball BA.Low-level production of the superoxide anion (O2*-) is an important signal transduction event in sperm function including capacitation; however, excessive production of O2*- can be detrimental to sperm function. The objective of this study was to assess dihydroethidium (DHE) as a probe for O2*- in equine spermatozoa. Ejaculated spermatozoa were separated by centrifugation over a Percoll gradient (40:80), and loaded with DHE (2.0 microM) as well as with calcein-acetoxymethylester (CAM, 7.8 nM) to determine cell viability. In Experiment 1, cells were incubated with the xanthine-xanthine oxidase ...
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