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Topic:Biochemistry
The study of biochemistry in horses encompasses the chemical processes and substances that occur within equine organisms. This field investigates the molecular interactions and pathways that are fundamental to horse physiology, including metabolism, enzyme activity, and genetic expression. Key areas of interest include the examination of metabolic disorders, nutrient absorption, and the biochemical basis of muscle function and energy production. Researchers utilize biochemical analysis to understand health and disease mechanisms in horses, contributing to the development of diagnostic tools and therapeutic strategies. This page gathers peer-reviewed studies and scholarly articles that explore various biochemical processes and their implications for equine health and performance.
The mobile bag technique as a method for determining the degradation of four botanically diverse fibrous feedstuffs in the small intestine and total digestive tract of ponies. An in sacco mobile bag technique was used to determine the rate and extent of disappearance of unmolassed sugar-beet pulp, soyabean hulls, hay cubes and an oat hull-naked oats mixture (67:33, w/w) in the foregut and total digestive tract of ponies. Ponies were administered naso-gastrically polyester mesh bags containing 350 mg feed, in a Latin square design. Bags were collected at the ileo-caecal junction (small intestine bag, SIB) and in the faeces (faecal bag, FB) and their residues analysed for proximate constituents and NSP composition and content. DM disappearances from individual bags we...
Excessive degradation of type II collagen in articular cartilage in equine osteochondrosis. Articular osteochondrosis (OCD) occurs in both man and animals. The etiology remains to be determined. Studies of OCD lesions in animals may provide clues as to its pathogenesis. The aim of our study was to determine whether there was evidence for increased degradation namely proteoglycan (PG) release and type II collagen cleavage in articular cartilage harvested from OCD lesions. We examined ex vivo explants at post-mortem from equine OCD lesions and macroscopically normal site and age matched cartilage. These were cultured over a 10 day period in serum-free medium. Type II collagen cleavage ...
Dopaminergic and opioidergic regulation of gonadotropin and prolactin release in stallions. In the non-breeding season, LH release is reduced via dopaminergic systems in the ram. On the other hand, our previous studies demonstrated an opioidergic inhibition of LH release in stallions outside the breeding season. Thus, in the present study we investigated the dopaminergic regulation of LH and prolactin secretion in stallions, considering interactions between dopamine and opioids. To achieve this, stallions (n=8) were treated with the dopamine antagonist sulpiride (0.6 mg/kg), the opioid antagonist naloxone (0.5 mg/kg), sulpiride plus naloxone or saline in December, March and June. Two...
Calcium homeostasis and intact plasma parathyroid hormone during exercise and training in young Standardbred horses. Physical exercise is known to affect calcium homeostasis in horses, but there is little information on the hormonal regulation of calcium metabolism during exercise. In order to evaluate the effects of exercise and training on calcium homeostasis and intact plasma parathyroid hormone, 7 untrained Standardbred horses were studied in a 6 week training programme. These horses were accustomed to running on the treadmill 3 weeks before onset of training and were exercised on a high-speed treadmill with an initial incremental standardised exercise test (SET 1: 6 incremental steps of 5 min duration e...
Expression of a 4-(hydroxy-3-nitro-phenyl) acetyl (NP) specific equi-murine IgE antibody that mediates histamine release in vitro and a type I skin reaction in vivo. Due to characteristic clinical signs, immunoglobulins of isotype E (IgE) are believed to be involved in several allergic diseases of the horse. To date, closer investigations have been hampered by the fact that neither purified equine IgE nor anti-equine IgE monoclonal antibodies were available for IgE isotype determination. As an approach to solve this problem, we constructed a stable cell line (EqE6) that expresses recombinant equi-murine IgE specific for 4-(hydroxy-3-nitro-phenyl) acetyl (NP). Biochemical analysis of the purified protein revealed a highly glycosilated IgE monomer of approxi...
Effect of (auto)phosphorylation on the kinetic behavior of the Ca2+/calmodulin-dependent protein kinase II from horse brain. After extraction and purification, the kinetic behavior of the Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) from horse brain was investigated as a function of ATP and a synthetic substrate, syntide-2. Both phospho- and dephospho- forms of the enzyme obey a bi-bi random mechanism. The K(M)s for ATP (K(M,ATP)) and syntide-2 (K(M,syntide-2)) were determined as equal to 80 and 30 microM, respectively. However, the maximum reaction yield is decreased by 50% when the enzyme is (auto)phosphorylated. In addition, this phosphorylated form of the enzyme leads to the formation of a total...
Biochemical markers of bone activity in young standardbred horses during different types of exercise and training. Seven untrained Standardbred horses were used in a training programme of 6 weeks to evaluate the effects of exercise and training on bone metabolism. The horses were exercised on a treadmill according to a standardized exercise test (SET 1: six incremental steps, 5 min duration each; start 5 m/s, increase 1 m/s). SET 1 was followed by a training programme of 6 weeks. In alternating order: high-speed exercise (HSE): 15 min duration, start at VLa4, continuous increase in speed every 60 s by 0.3 m/s (14 incremental steps); low-speed exercise (LSE): constant velocity at VLa2.5, duration: approxima...
Appearance of nitrite reducing activity of cytochrome c upon heat denaturation. The appearance of NO2- reducing activity of cytochrome c (Cyt c) upon heat denaturation was investigated with equine heart Cyt c. Denatured equine heart Cyt c (dCyt c), which was treated at 100 degrees C for 30 min, had NO2- reducing activity in the presence of dithionite and methylviologen in an aqueous solution under anaerobic conditions. In contrast, hemoglobin and myoglobin had no such activity under the same conditions. Using spectroscopic methods, we found that the appearance of this activity in the Cyt c was due to the following intramolecular changes: unfolding of the peptide chain, ex...
Serum fluoride concentrations, biochemical and histopathological changes associated with prolonged sevoflurane anaesthesia in horses. The volatile anaesthetic sevoflurane is degraded to fluoride (F-) and a vinyl ether (Compound A), which have the potential to harm kidney and liver. Whether renal and hepatic injuries can occur in horses is unknown. Cardiopulmonary, biochemical and histopathological changes were studied in six healthy thoroughbred horses undergoing 18 h of low-flow sevoflurane anaesthesia. Serum F- concentrations were measured and clinical laboratory tests performed to assess hepatic and renal function before and during anaesthesia. Necropsy specimens of kidney and liver were harvested for microscopic examinat...
Binding of radiolabeled porcine motilin and erythromycin lactobionate to smooth muscle membranes in various segments of the equine gastrointestinal tract. To identify and characterize motilin receptors in equine duodenum, jejunum, cecum, and large colon and to determine whether erythromycin lactobionate competes with porcine motilin for binding to these receptors. Methods: Specimens of various segments of the intestinal tracts of 4 adult horses euthanatized for reasons unrelated to gastrointestinal tract disease. Methods: Cellular membranes were prepared from smooth muscle tissues of the duodenum, jejunum, pelvic flexure, and cecum. Affinity and distribution of motilin binding on membrane preparations were determined by use of 125I-labeled synth...
Effects of an adenosine kinase inhibitor and an adenosine deaminase inhibitor on accumulation of extracellular adenosine by equine articular chondrocytes. To investigate accumulation of extracellular adenosine (ADO) by equine articular chondrocytes and to compare effects of adenosine kinase inhibition and adenosine deaminase inhibition on the amount of nitric oxide (NO) produced by lipopolysaccharide (LPS)-stimulated chondrocytes. Methods: Articular cartilage from metacarpophalangeal and metatarsophalangeal joints of 14 horses. Methods: Chondrocytes were cultured as monolayers, and cells were incubated with LPS, the adenosine kinase inhibitor 5'-iodotubercidin (ITU), or the adenosine deaminase inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine hydro...
Stabilization of protein by replacement of a fluctuating loop: structural analysis of a chimera of bovine alpha-lactalbumin and equine lysozyme. Equine lysozyme is a calcium-binding lysozyme and an evolutional intermediate between non-calcium binding c-type lysozyme and alpha-lactalbumin. We constructed a chimeric protein by substituting the fluctuating loop of bovine alpha-lactalbumin with the D-helix of equine lysozyme. The substitution affects the protection factors not only in the fluctuating loop but also in the antiparallel beta-sheet, the A- and B-helices, and the loop between the B-helix and the beta-sheet. Amide protons in these regions of the chimera are more protected from exchange than are those of bovine alpha-lactalbumin....
Progesterone induces acrosome reaction in stallion spermatozoa via a protein tyrosine kinase dependent pathway. Progesterone (P(4)) is a physiological inducer of the acrosome reaction (AR) in stallion spermatozoa. However, the capacitation-dependent changes that enable progesterone binding, and the nature of the signaling cascade that is triggered by progesterone and results in induction of the AR, are poorly understood. The aim of the current study was, therefore, to investigate the protein kinase dependent signaling cascades involved in progesterone-mediated induction of the AR in stallion spermatozoa. In addition, we aimed to determine whether bicarbonate, an inducer of sperm capacitation, acted via ...
Production of amines in equine cecal contents in an in vitro model of carbohydrate overload. Acute laminitis can be induced experimentally in horses by the administration of carbohydrate, resulting in fermentation within the cecum and ischemia-reperfusion of the digits. The products of fermentation that trigger acute laminitis are as yet unknown; however, compounds such as amines might play a role due to their potential vasoactive properties. The objectives of this study were to quantify the amines present in equine cecal contents and to use a model of carbohydrate overload in vitro to test the hypothesis that carbohydrate fermentation is associated with increased amine production. Ce...
Comparative study of tyrosine radicals in hemoglobin and myoglobins treated with hydrogen peroxide. The reactions of hydrogen peroxide with human methemoglobin, sperm whale metmyoglobin, and horse heart metmyoglobin were studied by electron paramagnetic resonance (EPR) spectroscopy at 10 K and room temperature. The singlet EPR signal, one of the three signals seen in these systems at 10 K, is characterized by a poorly resolved, but still detectable, hyperfine structure that can be used to assign it to a tyrosyl radical. The singlet is detectable as a quintet at room temperature in methemoglobin with identical spectral features to those of the well characterized tyrosyl radical in photosystem...
Myosin heavy chain-based fibre types in red cell hyper- and normovolaemic Standardbred trotters. An assumed link between red cell hypervolaemia, an excessive amount of training and impaired performance of hypervolaemic horses has led to a theory that the muscle fibres could be affected. Myosin heavy chain (MHC)-based fibre type composition in gluteus medius muscle of red blood cell normo- (NV) and hypervolaemic (HV) Standardbred trotters was evaluated using immunohistochemistry. Muscle biopsies were obtained from 13 NV and 16 HV horses. Serial transverse sections were cut and reacted with antibodies against different isoforms of the myosin heavy chains MHCI, MHCIIA and MHCIIX. Sections we...
Phosphorylation of protein tyrosine residues in fresh and cryopreserved stallion spermatozoa under capacitating conditions. Phosphorylation of tyrosine residues on sperm proteins is one important intracellular mechanism regulating sperm function that may be a meaningful indicator of capacitation. There is substantial evidence that cryopreservation promotes the capacitation of sperm and this cryocapacitation is frequently cited as one factor associated with the reduced longevity of cryopreserved sperm in the female reproductive tract. This study was designed to determine whether stallion sperm express different levels of tyrosine phosphorylation after in vitro capacitation and whether thawed sperm display similar ph...
Lactate transport in red blood cells by monocarboxylate transporters. The lactate transport activity of red blood cells (RBC) varies widely among different species; in equine RBC, the activity of the main lactate carrier, H+-monocarboxylate co-transporter (MCT), is distributed bimodally. The influence of lactate transport activity is measurable in vivo; after maximal exercise, the RBC lactate concentration in horses with high (HT) lactate transport activity is higher than in those with low (LT) activity. To study the expression of MCT in HT and LT horses, blood samples were taken from 10 horses at rest and after submaximal exercise. Blood and plasma lactate conc...
Effects of prolonged training, overtraining and detraining on skeletal muscle metabolites and enzymes. Thirteen Standardbred horses trained intensively for 34 weeks and detrained for 12 weeks to investigate the effects of training, overtraining and detraining on muscle metabolites, buffering capacity and enzyme activities (CS, HAD and LDH). After a standardised exercise test to fatigue at 10 m/s (approximately 100% VO2max), there was significant depletion of [ATP], [PCr] and muscle [glycogen] and accumulation of muscle and plasma [lactate], [NH3] and elevated muscle temperature. After training, associated with increased run time to fatigue (148%), there was reduced depletion of muscle [glycogen...
Plasma lactate and uric acid responses to racing in pacing Standardbreds and relationships with performance. Relationships between racing performance and plasma lactate and uric acid concentrations after racing were investigated in pacing Standardbred racehorses. The study was undertaken in order to better understand factors that limit racing performance. Twenty horses were tested after races of 1760 m and 28 horses after races over 2160 m. Blood samples were taken 30-60 min before and 8 and 30 min after a race. There were no significant differences between the race distances for prerace and 8 min postrace plasma lactates. Significant low correlations were obtained for plasma lactate concentration 8 ...
Structural and oxidative enzyme characteristics of the diaphragm. During exercise, the horse can achieve oxygen uptakes and ventilations in excess of 200 ml/kg/min and 1800 l/min, respectively. Whether the diaphragm has the capacity to contribute substantially to inspiratory effort in the exercising horse is not known. To investigate the potential for the horse diaphragm to generate tension, lung displacement and sustain ventilatory function, we measured diaphragm thickness, muscle length and oxidative enzyme activity (citrate synthase) within the ventral, medial and dorsal costal and crural diaphragm. In the diaphragms of 6 mature horses (5 Thoroughbreds, o...
Training-induced alterations in glucose metabolism during moderate-intensity exercise. In several species, physical conditioning (training) provokes a large shift in substrate utilisation during submaximal exercise. Few studies in horses have quantitatively examined these effects. Therefore, the effects of exercise training on plasma glucose kinetics during submaximal exercise were examined in 7 horses (5 Thoroughbred, 2 Standardbred; age 3-9 years) that had been paddock-rested for at least 6 months. Two days after determination of maximum aerobic capacity (VO2max), horses ran on a treadmill (4 degree incline) at 55% of VO2max (UT) for 60 min or until fatigue and then completed ...
Effects of fat adaptation on glucose kinetics and substrate oxidation during low-intensity exercise. This study was designed to determine the effects of fat adaptation on carbohydrate and fat oxidation in conditioned horses during low-intensity exercise. Five mature Arabians were studied. The study was conducted as a crossover design with 2 dietary periods, each of 10 week's duration: a) a control (CON) diet, and b) a fat-supplemented (FAT) diet. The total amount of digestible energy (DE) supplied by the fat in the CON and FAT diets was 7% and 29%, respectively. During each period, the horses completed exercise tests at the beginning of the period (Week 0) and after 5 and 10 weeks on the diet...
Pro- and macroglycogenolysis in skeletal muscle during maximal treadmill exercise. The purpose was to investigate the degradation of proglycogen and macroglycogen in skeletal muscle during intense exercise. Ten Standardbred trotters performed a maximal treadmill exercise test comprising a warm-up period, an exercise period, starting at 7 m/s with increments of 1 m/s every 60 s until the onset of fatigue (mean +/- s.d. 246 +/- 32 s) and a walking recovery period. Muscle biopsies were taken at rest, immediately after exercise and 15 min postexercise. The exercise caused a marked anaerobic metabolism as shown by the decrease in both muscle ATP and creatine phosphate and increas...
Changes in skeletal muscle GLUT4 content and muscle membrane glucose transport following 6 weeks of exercise training. This study examined changes in skeletal muscle GLUT4 content and glucose transport in isolated muscle membranes (GT) from horses before and 2 min after standardised submaximal exercise tests (SET) prior to and after completion of 6 weeks of training. Seven horses, age 3-9 years, body mass mean +/- s.e. 530 +/- 19 kg, and sedentary for at least 4 months, completed 6 weeks of training on a treadmill. An initial SET (UT) was performed on a 4 degree incline at a speed equivalent to 55% of pretraining VO2max and was repeated post-training at the same absolute workload (ABS). A third SET (REL) was p...
Inhibition of articular cartilage degradation by glucosamine-HCl and chondroitin sulphate. Glucosamine and chondroitin sulphate in many animal and human trials has improved joint health. In vitro studies are beginning to clarify their mode of action. The objective of this research was to: 1) determine at what concentrations glucosamine-HCl (GLN) and/or chondroitin sulphate (CS) would inhibit the cytokine-induced catabolic response in equine articular cartilage explants and 2) to determine if a combination of the 2 was more effective at inhibiting the catabolic response than the individual compounds. Articular cartilage was obtained from carpal joints of horses (age 1-4 years). Carti...
Potassium and lactate uptake by noncontracting tissue during strenuous exercise. Efflux of potassium (K+) and lactate (lac-) from active skeletal muscle during high intensity exercise leads to increased plasma [K+] and [lac-] in venous and arterial blood. The exercise-induced increases in these ions in human athletes is ameliorated, in part, by uptake by noncontracting tissue, reflected by an increase in the arteriovenous difference across these tissues from rest to exercise. To test the hypothesis that these ions are taken up by noncontracting tissue in strenuously exercising horses, plasma [K+] and [lac-] were measured in blood samples collected simultaneously from the c...
Equine endurance exercise alters serum branched-chain amino acid and alanine concentrations. Six 2-year-old Arabian horses were used to determine whether 60 km prolonged endurance exercise (approximately 4 h) alters amino acid concentrations in serum and muscle, and the time required for serum amino acid concentrations to return to basal resting values. Blood and muscle samples were collected throughout exercise and during a 3 day recovery period. Isoleucine concentration in muscle tended to increase and leucine and valine did not change due to exercise. Serum alanine concentrations did not increase immediately after exercise, but increased at 24, 48 and 72 h postexercise. Serum isole...
Effect of dietary lipid on response to exercise: relationship to metabolic adaptation. The aim of the present study was to relate changes in muscle oxidative capacity and free fatty acid flux in response to oil supplementation to fuel utilisation during subsequent exercise of varying intensities. Following 10 weeks of oil supplementation there was an increased capacity for fat utilisation during low and moderate intensity exercise as indicated by a lower respiratory exchange ratio (RER) (P<0.05). We suggest that this was contributed to by a parallel increase in the oxidative capacity of muscle as indicated by a significant increase in the activity of muscle citrate synthase (...
