Equine metabolism encompasses the biochemical processes that occur within horses to maintain life, including the conversion of food into energy, the synthesis of necessary compounds, and the elimination of waste products. These processes are essential for supporting various physiological functions such as growth, reproduction, and physical performance. Key components of equine metabolism include carbohydrate, fat, and protein metabolism, each of which contributes to the overall energy balance and health of the horse. Factors influencing metabolic rate and efficiency in horses include age, breed, diet, exercise, and health status. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, regulation, and implications of metabolic processes in equine physiology.
Bisgard GE, Forster HV, Byrnes B, Stanek K, Klein J, Manohar M.Ventilation, metabolism, arterial blood gases, and blood and cerebrospinal fluid (CSF) acid-base status were measured in exercise studies on seven ponies during mild, moderate, and near-maximal treadmill exercise. CSF and arterial blood were sampled via indwelling catheters. Generally measurements were made during the 3rd, 6th, and 9th minute of steady-state exercise, with CSF sampled only during the 9th minute. Alveolar ventilation (VA) and metabolic rate (VO2) increased proportionately during exercise below the anaerobic threshold, but above this threshold, VA increased at a faster rate than...
Powis G, Snow DH.There are now several examples showing that experimentally induced changes in hepatic blood flow can have a marked effect upon the elimination of certain high clearance drugs. Changes in hepatic blood flow produced by exercise might therefore be expected to influence the clearance of these drugs. There was an increase of up to 100%, compound to control values, in the plasma levels of propranolol in horses given either d- or dl-propranolol, 0.2 mg/kg b.wt., and then subjected to sustained exercise for 30 minutes. There was, however, no similar increase with exercise in the plasma levels of anti...
Kohn CW, Muir WW, Sams R.Plasma volume and extracellular fluid volume were determined in horses at rest and immediately after a standardized exercise test. Exercising during training demonstrated no detectable effect on plasma or extracellular fluid volume determinations when compared with resting values. An explanation of this finding and an argument for the proper analysis of plasma and extravascular fluid volumes are presented.
Chand N, Eyre P.Histamine, 2-methylhistamine (a specific H1-agonist), 5-HT, PGF2alpha, SRS-A, bradykinin (BK) and carbachol contract bronchial and tracheal smooth muscles of the horse. Isoprenaline, PGE1, E2, dimaprit and 4-methylhistamine (last two = specific H2-agonists) relaxed airways which were partially contracted to carbachol. Mepyramine (a specific H1-antagonist) selectively antagonized contractions to histamine. In the presence of mepyramine, histamine caused relaxation of airways partially contracted to carbachol. Metiamide and burimamide (specific H2-antagonists) specifically antagonized or reverse...
Jondorf WR, Moss MS.1. Cross-bred and thoroughbred geldings were injected with veterinary doses of various synthetic anabolic steroids. Urines collected sequentially from treated animals were analysed, following solvent extraction, by radioimmunoassay using 19-[3H]nortestosterone and an antibody raised against a 19-nortestosterone immunogen. 2. Urinary excretion of 19-nortestosterone and/or its cross-reacting metabolites was detectable for various times after administration of different nortestosterone esters, as follows: phenylpropionate (400 mg), greater than 14 days; cyclohexylpropionate (100 mg), greather tha...
Schryver HF, Hintz HF, Lowe JE.The effect of exercise on the pathways of metabolism of 40Ca and 47Ca was studied in 4 yearling Standardbred horses in 4 consecutive treatment periods: (1) no exercise, (2) trotting 16 km/day, (3) trotting 10 km/day, and (4) no exercise. Metabolic balance studies and studies of 47Ca kinetics were conducted during the final week of each month-long treatment period. The urinary excretion of 40Ca and 47Ca decreased 50% to 75% during the exercise periods. Retention of 47Ca increased during the exercise periods, but the retention of dietary Ca (40Ca) did not change. The efficiency of Ca absorption ...
Joppich-Kuhn R, Luisi PL.The interaction between horse liver alcohol dehydrogenase and the oxidized and reduced forms of the 3-thionicotinamide--adenine dinucleotide coenzyme analogues (sNAD and sNADH) has been investigated by ultraviolet absorption, fluorescence and circular dichroism. The fluorescence of sNADH is enhanced when bound to the enzyme, and the protein fluorescence is quenched by both sNADH (60--65%) and sNAD (65%). The possible origin of the larger quenching produced by sNAD with respect to that of NAD is discussed. Coenzyme dissociation constants have been determined by monitoring the quenching of the p...
Caple IW, Edwards SJ, Forsyth WM, Whiteley P, Selth RH, Fulton LJ.The activity of glutathione peroxidase, a selenium containing enzyme, was measured in the blood of horses to determine its usefulness as an indicator of selenium status. In 15 horses the enzyme activity was positively related to the blood selenium concentration (P less than .001, r-0.98) over the range of enzyme activities of 8.2 to 140 units (mumoles NADP-oxidised/min/gHb) and selenium concentrations of 0.24 to 2.74 mumol/l. In a group of 8 horses which 2 foals had died with lesions of muscular dystrophy the enzyme activity increased from a mean of 11.8 units before treatment with selenium to...
Houghton E, Oxley GA, Moss MS, Evans S.A method is described to confirm the presence of 19-nortestosterone metabolites in urine after the administration of veterinary preparations of this anabolic steroid to horses. The method is based upon the detection, by gas chromatography mass spectrometry or selected ion monitoring, of an isomer of estrane-3,17-diol in the urine.
Stefani M, Berti A, Camici G, Manao G, Cappugi G, Ramponi G.Horse muscle acylphosphatase consists of a main chain S-S bound to glutathione. It was found that removal of the glutathione by reduction and successive carboxymethylation of the only cysteine of the main chain affects the stability of the enzyme, mainly with respect to thermal inactivation. On the other hand, the kinetic properties of the enzyme are affected very little.
Pine L, Reeves MW.Various sugars were tested for their effect on the differential rate of synthesis of M protein during the growth of Streptococcus pyogenes strain 0055 M12T12. In a semisynthetic medium alone, a high rate of M protein synthesis occurred with glucose as a substrate; decreasing rates of synthesis occurred with sucrose and trehalose, in that order, although the rates of growth were approximately equal with all sugars. A period of derepressed synthesis of M protein occurred in the lag phase of growth and in the stationary period as the substrates were being depleted. Although glucose inhibited the ...
Tax WJ, Veerkamp JH.1. Activities of ADA and PNP were measured in erythrocytes and lymphocytes of man, horse and cattle. 2. In bovine hemolysates both enzyme activities are low when compared with activities in human hemolysates. In horse hemolysates both enzyme activities are virtually absent. 3. Enzyme activities are consistently lower (about 50%) in intact lymphocytes than in sonicated lymphocytes. This finding suggests that the uptake of nucleosides is rate-limiting for both enzymes in intact lymphocytes. 4. The activity of ADA in horse lymphocytes is comparable to that in lymphocytes of patients with severe c...
Houghton E.1. The metabolism of 19-nortestosterone in a cross-bred horse has been studied using 14C-labelled material. 2. Two neutral metabolites isolated from urinary extracts by column chromatography were identified as isomers of 3-hydroxyestran-17-one and estrane-3,17-diol by g.l.c.-mass spectrometry. 3. The stereochemistry of the two metabolites has been investigated by comparison of the retention times of their trimethylsilyl derivatives with those of standard steroids of known configuration.
Franken P, Schotman JH.The activities and concentrations of a number of erythrocytic enzymes and intermediate products of erythrocyte metabolism were determined in twenty-one normal standard-bred horses which were studied clinically and biochemically. These studies showed that equine anaerobic glycolysis is characterized by a biochemical pattern similar to that observed in human PK deficiency. The greater sensitivity of equine haemoglobin to oxidants is attributable either to low stability of GSH, which may be due either to the low activity of GR or that of 6PGD as observed in the studies. In addition, the saturatio...
Muir WW, Sams RA, Ashcraft S.The hemodynamic, respiratory, and behavioral effects, as well as the pharmacokinetic properties of methocarbamol, were determined in horses. Heart rate, cardiac output, arterial and venous blood pressures, respiratory rate, and arterial blood gases did not change after IV methocarbamol (4.4, 8.8, 17.6 mg/kg) administration. There were no signs of behavior modification or ataxia observed. Analysis of plasma concentration time data indicated that the disposition of methocarbamol may be dose-dependent. Clearance and steady-state volume of distribution decreased as the dose increased. Plasma conce...
Kowalski J, Roberts A, Williams J, Hintz HF, Daniluk P, Schryver HF.Three pony geldings were given sodium bicarbonate orally in order to study the effect on blood pH and bicarbonate and to determine if frequency of dosing influences the response. In a preliminary study, it appeared that a carry-over effect might occur if the interval between dosing was only 2 days. The ponies received 2 doses of sodium bicarbonate (400 mg/kg) 7 days apart in trial one and then in trial two they received 2 doses of sodium bicarbonate 4 days apart. The sodium bicarbonate was mixed with 2 liters of warm water and given through a nasogastric tube on each trial day. Blood samples w...
Filipović N, Stojević Z, Plevnik N, Mašek T, Prvanović N, Tuček Z.The concentrations of carboxyterminal cross-linked telopeptide of type I collagen (CTx), osteocalcin (OCN), parathyroid hormone (PTH) and insulin-like growth factor (IGF-I) were measured in the blood serum of two age groups of mares during late pregnancy (70-50 days before foaling), during early lactation and at the peak of lactation (10-30 and 55-80 days after foaling, respectively). During late pregnancy, the PTH was higher in older (8-19 years old), compared to younger animals (3.5-4 years old) (P < 0.05). The OCN was higher in younger group during late pregnancy (P < 0.05). IGF-I was...
Fenton JI, Orth MW, Chlebek-Brown KA, Nielsen BD, Corn CD, Waite KS, Caron JP.The effect of longeing and glucosamine supplementation on known biological markers of joint disease was studied in yearling quarter horses. Twenty-one yearling quarter horses were randomly assigned to one of 4 treatments: 1) longeing (longeing 20 min daily) supplement control (LN); 2) longeing/glucosamine (LG); 3) walking (mechanical walker for 120 min daily (WN)); and 4) walking/glucosamine (WG). Oral glucosamine was administered at 5.5 g b.i.d. weeks 1-4, 3.5 g b.i.d. during weeks 5-6, and 2.0 g b.i.d. during weeks 7-8. Serum was obtained weekly for 8 wk and analyzed for keratan sulfate and ...
Turk MA, Thomas DE.To study the role of cytochrome P-450-dependent mixed function oxidase reactions in equine 3-methylindole (3MI) toxicosis, ponies were given 20 mg of phenobarbital/kg of body weight at 72, 60, 48, 36, and 24 hours before 100 mg of oral 3MI/kg to induce cytochrome P-450 or no treatment (controls). Maximal 3MI plasma concentration was decreased and clearance was faster in phenobarbital-treated ponies. Plasma 3MI was still detectable 12 and 36 hours after dosing in phenobarbital-treated and control ponies, respectively. Phenobarbital treatment induced a distribution phase with transition from a 1...
Martens RJ, Malone PS, Brust DM.Oral lactose tolerance tests were evaluated in 25 healthy foals (principals) assigned to 4 groups of approximately 1 week, 4 weeks, 8 weeks, and 12 weeks of age. Lactose monohydrate (1 g/kg of body weight [in a 20% water solution]) was administered via nasogastric tube after a 4-hour fast. Plasma glucose concentrations were monitored before dosing (0 minutes) and sequentially for 300 minutes. Six control foals were given a volume of water equivalent to the volume of lactose monohydrate administered to principal foals. After oral lactose loading, mean plasma glucose concentrations of all princi...
Zeyner A, Hoffmeister C, Einspanier A, Gottschalk J, Lengwenat O, Illies M.Quarter Horses are particularly susceptible to polysaccharide storage myopathy (PSSM). Nutritional therapy and possibly prophylaxis includes fat-supplemented diets whilst starch supply should be kept to a minimum. Objective: To investigate the glycaemic and insulinaemic response of clinically normal Quarter Horses to concentrates high in fat and low in starch. Methods: Twelve Quarter Horses were studied. The precondition for inclusion in the study population was that the horses had not shown clinical signs of myopathy. The Quarter Horses were fed according to a 4 x 4 Latin square design haylag...
Díez E, López I, Pérez C, Pineda C, Aguilera-Tejero E.Donkeys appear to be more predisposed than large breed horses to suffer from hyperlipemia. The reason for that predisposition is unknown but anorexia is a consistent feature of the disease. Leptin, a protein synthesized in fat tissue, is one of the major inhibitors of appetite in mammals. Objective: We hypothesized that donkeys could have elevated plasma leptin concentrations compared to horses. Methods: Blood samples were obtained from 50 donkeys for measurement of leptin, triglycerides (TGs), glucose, and insulin. Glucose/insulin ratio, modified insulin to glucose ratio, and reciprocal of th...
Brotherton HO, Yost RA.A screening and confirmation procedure for drugs and metabolites in the blood serum and urine of racing animals was developed. Equine blood serum was spiked with low concentrations of several drugs of interest. Canine blood serum and urine were collected following oral doses of diethylcarbamazine, procaine, and phenylbutazone. Serum, urine, and extracts of each were analyzed, using a triple quadrupole mass spectrometer. Simultaneous screening of up to 50 drugs was possible in a single sample, in less than 2 minutes. Detection limits for most compounds were in the ng/ml to microgram/ml range, u...
Frape DL.There is no clear evidence that the chronic requirement for any non-energy yielding nutrient rises in proportion as the energy requirement increases with hard work. The need for protein, and probably that for calcium, remain a function of bodyweight daily. Some proportionality with energy may exist for certain nutrients, although the evidence has not been adduced. For example, because of an increase in both the proportion and amount of propionic acid in the volatile fatty acids of caecal contents, the tissue requirement for vitamin B12 may rise with an increase in the rate of energy metabolism...
Turcott-White SK, Nielsen BD, Robison CI, Skelly CD, Rosenstein DS, Pritchard A, Herdt T.The objective of this study was to compare the feed intake and the apparent digestibilities of three different diets varying in concentrate-to-roughage ratios in weanling horses (n = 24) at 5 and 8 months of age. Horses were stratified by breed, gender, birth date and body weight and assigned to one of three dietary treatments containing the following concentrate-to-roughage ratios on an as-fed basis: 70:30 (High Con), 50:50 (Equal) and 30:70 (Low Con). All horses were fed their respective diets for a 10-day adaptation period and a 4-day collection period at 5 and 8 months. There were no d...
Ralston SL, Freeman DE, Baile CA.The roles of volatile fatty acids (VFA) and of the large intestine in the control of feeding in ponies were investigated. Ponies with cecal fistulas were adapted to ad libitum access to pelleted feed. Treatment solutions were given as a bolus 15 min before the animals were allowed free access to feed after a 4-h fast. Each dose of VFA solution was tested in a crossover design with a water control. When the ponies were permitted to eat after the treatments, the latency to eat, first meal size, and duration and first intermeal interval were recorded. Feed intakes were measured at 3 and 18 h afte...
Maxfield M, Celaya R, Andrade E, Rodrigues MAM, Santos AS, Ferreira LMM.The assessment of feed intake in stabled horses is a difficult task to accomplish. Faecal markers, namely n-alkanes, have been used successfully for the estimation of this important nutritional parameter. This usually involves the dosing of synthetic n-alkanes via different matrices, a laborious task that may also influence the animal normal foraging behaviour. An experiment was conducted to evaluate a relative simple methodology to quantify feed intake in horses, based on the provision of measured amounts of a concentrate supplement labelled with beeswax and the utilisation of n-alkanes as fa...
Bertone AL, Toofanian F, Stashak TS.Lactase, maltase, sucrase, and alkaline phosphatase activities were determined in the intestinal mucosa from 3 locations in the small intestine and 4 locations in the large intestine 1 year after extensive large-colon resection (group 1; n = 5) and 1 year after sham operation (group 2; n = 3) in horses. Lactase, maltase, and sucrase activities were similar (P greater than 0.05) between group-1 and group-2 horses in all locations measured in the intestinal tract. Alkaline phosphatase activity in the remaining large colon of group-1 horses was significantly (P less than 0.05) greater than the ac...
Karatt TK, Sathiq MA, Laya S, Philip M, Karakka Kal AK, Subhahar MB.An effective alternative to testosterone therapy is selective androgen receptor modulators, a class of compounds that has a tissue-specific effect on muscle and bone. These drugs, which enhance performance, pose a severe abuse risk in competitive sports. GLPG0492 is one of the selective androgen receptor modulators discovered in recent decades. This compound has a unique tissue-specific action for muscle and bone against steroid receptors and acts as a partial agonist for androgen receptors. This study examined GLPG0492 and its metabolites in vitro using equine liver microsomes. Liquid chromat...
Krumrych W, Skórzewski R, Malinowski E.The aim of this study was to determine the effect of duration and temperature of sample storage on whole blood chemiluminescence measurement results. Venous blood from 18 clinically healthy Polish half-bred horses aged 4 to 11 years were used in the study. Luminol dependent chemiluminescence (CL) was used to measure neutrophil oxygen metabolism in whole blood. Blood samples were examined for spontaneous CL and stimulated by a surface receptor stimulus as well as extra-receptor stimulus. The assay was performed in two parallel experimental sets with samples stored at 4 and 22 °C, respectively....
Kramer KJ, Kagan IA, Lawrence LM, Goff BM, Smith SR.An excess of water-soluble carbohydrates (WSCs) can present a risk for horses prone to pasture-associated laminitis or some other metabolic conditions. Determining WSC concentrations in commonly grazed cool-season grasses, at different times of day and under different fertilization treatments, can help optimize grazing times and management strategies. The goals of this study were to develop a near-infrared reflectance spectroscopy (NIRS) calibration curve for WSC and to apply it to four cool-season grass species. Tall fescue, Kentucky bluegrass, orchardgrass, and perennial ryegrass (10 cultiva...
McKeever KH.This article has presented information on the importance of the sympathetic nervous system in the response to exercise. The authors have reviewed the very limited information on the effects of sympathomimetic and sympatholytic drugs on exercise performance in the horse. Most of these drugs are specifically prohibited under the rules of racing, and they have significant side effects that either decrease performance or make their use dangerous to both the horse and horse-man. Additionally, all of these drugs or their metabolites are readily detected by current drug testing protocols. Further inf...
Pilarczyk B, Tomza-Marciniak A, Stankiewicz T, Błaszczyk B, Gaczarzewicz D, Smugała M, Udała J, Tylkowska A, Kuba J, Cieśla A.The aim of this study was to determine serum selenium concentrations in Polish Konik horses residing in the Odra Delta Nature Park (Poland) and to evaluate the activity of glutathione peroxidase and Se content in testes of this horse breed. In over 95% of cases, serum Se concentration was below the optimal range, and none of the horses examined was deficient in this trace element. The lack of Se deficiency in the animals examined suggests however, that the Polish Konik horses have a natural ability to the optimal use of nutrients available in their life area. Testicular content of Se and GSHPx...
Ralston SL, Baile CA.The hypothesis that changes in blood glucose concentrations and(or) utilization rate affect feeding behavior in ponies was tested. Ponies (n = 4) were fasted for 4 h, then given iv injections of 50% glucose [.2 g glucose/BW.75 kg (low dose, LD) or 1.0 g glucose/BW.75 kg (high dose, HD)] or an equal volume of normal saline (HDS; LDS) 5 min before being allowed access to pelleted feed. Blood samples were drawn at regular intervals pre- and post-treatment and analyzed for plasma glucose (PG), immunoreactive insulin (IRI) and glucagon concentrations. All glucose-treated animals immediately ate mea...
Ohya T, Kondo T, Yoshikawa Y, Watanabe K, Orino K.In mammal circulation, various ferritin-binding proteins (FBPs) are thought to be involved in the clearance of circulating ferritin after complex formation with it. However, horse FBPs are known to cause inhibitory effects on ferritin immunoassay due to the concealment of the ferritin molecule to anti-ferritin antibodies used in the ferritin immunoassay. These inhibitory effects are eliminated by heat treatment of horse serum at 75°C for 15 min. The inhibitory effects on ferritin immunoassay in the sera of ten foal sera (5 females and 5 males) from 1 to 18 months were detected during all peri...
Chen CL, Pattison ML, Engleking LR, Gronwall RR.The effect of a subcutaneous injection of estradiol on the secretion of pituitary prolactin in the rat and the relationship between serum estradiol level and luteinizing hormone (LH) secretion in mare were reviewed. In addition, the effect of estradiol injection on LH secretion and the metabolism of [14C] estradiol in intact and bile duct fistulated pony mares were studied. Low (0.1 mug/day/rat) to moderate dose (5 mug/day/rat) of estradiol benzoate injected subcutaneously to mature or immature rats significantly increased pituitary content of prolactin and serum prolactin level five- to tenfo...
Johnson PJ.This article presents a brief overview of the physiology of homeostasis in the horse and discusses the units of measurement by which dissolved electrolytes are assessed. Differences in electrolyte composition between compartments of the body and the manner in which water is exchanged between them are reviewed. The mechanisms by which alterations in osmolality and effective circulating volume are detected and the physiological responses to those alterations are presented.
Beech J, Fletcher JE, Erwin K, Lindborg SR.To determine sensitivity of equine skeletal muscle to tetrodotoxin and compare that with sensitivity of murine and human skeletal muscles. Methods: Semimembranosus, vastus lateralis, triceps brachii, and masseter muscle specimens from 22 euthanatized horses, vastus lateralis muscle biopsy specimens from 25 clinically normal humans, and diaphragmatic muscle specimens from 6 mice. Methods: Electrically elicited twitch responses were measured in muscle specimens incubated in medium alone and with tetrodotoxin (100 nM, 400 nM, 1.6 microM for equine specimens and 100 nM, 200 nM, 400 nM, 800 nM, 1.6...
Ott EA, Brown MP, Roberts GD, Kivipelto J.Forty-four weanling horses were used in two experiments to evaluate the effect of starch intake on growth and skeletal development. In Exp. 1, the weanlings were fed either a grain-based, high-starch (31.1%, DM basis) concentrate or a by-product-based, low-starch (0.0%) concentrate with coastal bermudagrass (Cynodon dactylon) hay. Corn oil was used to equalize the energy concentration of the concentrates. The concentrate:hay ratio was 64:36 (as-fed basis), and intake was the same for both diets. Body weight gains were greater by the weanlings consuming the high-starch concentrate (0.81 vs. 0.6...
Luna SP, Taylor PM, Brearley JC.Glucose was infused intravenously into six ponies during halothane anaesthesia, to evaluate its effect on their endocrine response to anaesthesia. The ponies were premedicated with acepromazine, and anaesthesia was induced with thiopentone and maintained with halothane in oxygen for two hours. Glucose was infused to maintain the plasma glucose concentration above 20 mmol/litre. Anaesthesia was associated with hypothermia, a decrease in haematocrit, hypotension, hyperoxaemia, respiratory acidosis and an increase in the plasma concentrations of lactate and arginine vasopressin. The concentration...
Zuffa T, Bennet ED, Parkin TDH.Endurance riding is a popular equestrian sport organised at the international level by the Fédération Équestre Internationale (FEI). The sport involves prolonged exercise at speed over significant distances, which puts a substantial load on equine athletes' musculoskeletal systems and metabolism and contributes to the risk of injuries and other veterinary problems. The FEI employs a system of in-ride veterinary inspections aimed at early detection of such problems and elimination of horses unfit to continue in the ride. To date, risk factors for elimination due to irregular gait or metaboli...
Marr K, Marsh K, Hernandez L, Cunningham FM, Lees P.Leukotrienes, products of the 5-lipoxygenase pathway of arachidonic acid metabolism, possess properties consistent with their involvement in a range of inflammatory diseases. In this study the pharmacokinetics and pharmacodynamics of the selective 5-lipoxygenase inhibitor, fenleuton, have been examined in the horse. Orally administered fenleuton (four 5 mg kg(-1) doses, given once daily) was absorbed from the gastrointestinal tract, and penetrated readily into tissue cage exudate, the ratio of the plasma:exudate AUC0-48h being 0.90+/-0.02 (n=6). Ionophore-stimulated leukotriene (LT) B4 synthes...
Wong DM, Vo DT, Alcott CJ, Peterson AD, Brockus CW, Hsu WH.Arginine vasopressin (AVP) has received increased attention in equine critical care but there is minimal information of AVP concentration in foals. The clinical usefulness of measuring AVP in ill foals depends on knowledge of age-related changes in AVP concentrations in healthy foals. Objective: Plasma AVP concentrations will be significantly different when measured from birth to 3 months of age in healthy foals. Methods: Thirteen healthy university-owned foals. Methods: Prospective, observational study. Blood was collected from healthy foals at birth and 3, 5, 7, 10, 14, 21, 28, 42, 56, and 8...
Baracca A, Bucchi L, Ghelli A, Lenaz G.A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9-amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. We carried out a kinetic analysis of each enzyme complex studying the proton translocation and the electron transfer activities of complex I as well as the proton translocation and the ATP hydrolytic activities of complex V. A micromethod to prepare...