Search Results

Abstract

Objective—To investigate whether protein kinase C (PKC) isoforms are expressed in equine skeletal muscle and determine their distribution in various types of fibers by use of immunofluorescence microscopy.

Animals—5 healthy adult Dutch Warmblood horses.

Procedure—In each horse, 2 biopsy specimens were obtained from the vastus lateralis muscle. Cryosections of equine muscle were stained with PKC isoform (α, β1, β2, δ, ξ, or ζ)-specific polyclonal antibodies and examined by use of a fluorescence microscope. Homogenized muscle samples were evaluated via western blot analysis.

Results—The PKC α, β1, β2, δ, ξ, and ζ isoforms were localized within the fibers of equine skeletal muscle. In addition, PKC α and β2 were detected near or in the plasma membrane of muscle cells. For some PKC isoforms, distribution was specific for fiber type. Staining of cell membranes for PKC α was observed predominantly in fibers that reacted positively with myosin heavy chain (MHC)-IIa; PKC δ and ξ staining were more pronounced in MHC-I-positive fibers. In contrast, MHC-I negative fibers contained more PKC ζ than MHC-I-positive fibers. Distribution of PKC β1 was equal among the different fiber types.

Conclusions and Clinical Relevance—Results indicated that PKC isoforms are expressed in equine skeletal muscle in a fiber type-specific manner. Therefore, the involvement of PKC isoforms in signal transduction in equine skeletal muscle might be dependent on fiber type. ( Am J Vet Res 2004; 65:69–73)

Abstract

Objective—To investigate the effects of exercise on activation of mitogen-activated protein kinase (MAPK) signaling proteins in horses.

Animals—6 young trained Standardbred geldings.

Procedure—Horses performed a 20-minute bout of exercise on a treadmill at 80% of maximal heart rate. Muscle biopsy specimens were obtained from the vastus lateralis and pectoralis descendens muscles before and after exercise. Amount of expression and intracellular location of phosphospecific MAPK pathway intermediates were determined by use of western blotting and immunofluorescence staining.

Results—Exercise resulted in a significant increase in phosphorylation of p38 pathway intermediates, c-Jun NH2 terminal kinase (JNK), and heat shock protein 27 (HSP27) in the vastus lateralis muscle, whereas no significant changes were found in phosphorylation of extracellular regulated kinase. In the pectoralis descendens muscle, phosphorylation of p38 and HSP27 was significantly increased after exercise. Immunohistochemical analysis revealed fiber-type– specific locations of phosphorylated JNK in type 2a/b intermediate and 2b fibers and phosphorylated p38 in type 1 fibers. Phosphorylated HSP27 was strongly increased after exercise in type 1 and 2a fibers.

Conclusions and Clinical Relevance—The p38 pathway and JNK are activated in the vastus lateralis muscle after a single 20-minute bout of submaximal exercise in trained horses. Phosphorylation of HSP27 as detected in the study reported here is most likely induced through the p38 signaling pathway.

Abstract

Objective—To confirm whether the plasma glucose concentration curve obtained during oral glucose tolerance tests (OGTTs) in horses with equine motor neuron disease (EMND) is decreased, compared with that obtained in clinically normal horses, and determine whether that decrease is a result of defective glucose metabolism or intestinal glucose transport dysfunction.

Animals—8 horses with EMND and 44 matched control horses.

Procedure—Electromyography and OGTTs were performed in all 8 affected horses and 10 control horses. Intravenous GTTs (IVGTTs) were performed in 6 affected horses and another 11 control horses. The activity and levels of jejunal luminal membrane glucose transporter (Na⁺/glucose cotransporter isoform 1 [SGLT1]) were measured in 2 affected horses and 23 control horses.

Results—In horses with EMND, generalized neuropathy was detected via quantitative electromyography; the mean increase in plasma glucose concentration during the OGTT was significantly decreased, compared with the value in control horses. During the IVGTT, the mean increase in plasma glucose concentration was significantly lower than that of control horses. The activity and levels of SGLT1 in 2 affected horses were similar to those of control horses. Diagnosis of EMND was confirmed postmortem in all affected horses.

Conclusions and Clinical Relevance—Data suggest that the decreased plasma glucose curve obtained in horses with EMND during OGTTs (compared with control horses) is a result of overall enhanced glucose metabolism or abnormalities in the facilitated glucose transporters; definitive identification of the underlying mechanisms could aid in the development of appropriate treatments of EMND in horses. (Am J Vet Res 2005;66:93–99)

Abstract

OBJECTIVE To determine effects of anesthesia on plasma concentrations and pulsatility of ACTH in samples obtained from the cavernous sinus and jugular vein of horses.

ANIMALS 6 clinically normal adult horses.

PROCEDURES Catheters were placed in a jugular vein and into the cavernous sinus via a superficial facial vein. The following morning (day 1), cavernous sinus blood samples were collected every 5 minutes for 1 hour (collection of first sample = time 0) and jugular venous blood samples were collected at 0, 30, and 60 minutes. On day 2, horses were sedated with xylazine hydrochloride and anesthesia was induced with propofol mixed with ketamine hydrochloride. Horses were positioned in dorsal recumbency. Anesthesia was maintained with isoflurane in oxygen and a continuous rate infusion of butorphanol tartrate. One hour after anesthesia was induced, the blood sample protocol was repeated. Plasma ACTH concentrations were quantified by use of a commercially available sandwich assay. Generalized estimating equations that controlled for horse and an expressly automated deconvolution algorithm were used to determine effects of anesthesia on plasma ACTH concentrations and pulsatility, respectively.

RESULTS Anesthesia significantly reduced the plasma ACTH concentration in blood samples collected from the cavernous sinus.

CONCLUSIONS AND CLINICAL RELEVANCE Mean plasma ACTH concentrations in samples collected from the cavernous sinus of anesthetized horses were reduced. Determining the success of partial ablation of the pituitary gland in situ for treatment of pituitary pars intermedia dysfunction may require that effects of anesthesia be included in interpretation of plasma ACTH concentrations in cavernous sinus blood.

Abstract

OBJECTIVE To determine whether hypertension, high sympathetic tone, resting and exercising arrhythmias, and echocardiographic changes consistent with hypertensive cardiomyopathy were associated with equine metabolic syndrome (EMS) in ponies.

DESIGN Prospective case-control study.

ANIMALS 19 privately owned ponies with a diagnosis of EMS (history of laminitis, body condition score ≥ 7/9, cresty neck score ≥ 3/5, and abnormal oral sugar test result; cases) and 20 healthy control ponies.

PROCEDURES Heart rate (HR), noninvasively measured arterial blood pressure (BP), markers of autonomic tone (splenic volume and HR variability), 24-hour and exercising ECGs, and echocardiograms were compared between cases and controls.

RESULTS Compared with controls, cases had a higher mean ± SD HR (44.5 ± 7.5 beats/min vs 38.6 ± 6.8 beats/min) and median mean left ventricular wall thickness (2.0 cm vs 1.8 cm). No differences were identified between groups in BP, splenic volume, HR variability, and number of premature complexes in ECGs. Mean wall thickness was correlated with BP (r = 0.54), high-frequency power (r = −0.71), and ratio of low-to high-frequency power (φ = 0.66). Relative wall thickness was correlated with serum insulin concentration (r = 0.71).

CONCLUSIONS AND CLINICAL RELEVANCE Ponies with EMS had myocardial hypertrophy that was correlated with insulin response to an oral sugar test, sympathetic and parasympathetic tone, and BP. The heterogeneity and limited sample size of this preliminary study should be considered when drawing conclusions. Cardiovascular changes associated with this syndrome deserve further attention.

Abstract

Objective—To investigate the effects of acute exercise and long-term training on Na⁺,K⁺-ATPase content, mRNA isoforms, and protein concentration in equine muscle.

Animals—6 Standardbreds.

Procedures—Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na⁺,K⁺-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [³H]ouabain binding, real-time PCR assay, and western blotting, respectively.

Results—6 Na⁺,K⁺-ATPase mRNA isoforms were present in equine muscle, but only A2 and B1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms A1, A2, A3, and B2 in VLM and A1 and B3 in PDM. Training increased resting values for mRNA isoforms A3 and B1 in VLM and B3 in PDM. The Na⁺,K⁺-ATPase, [³H]ouabain binding, and proteins of mRNA A2 and B1 increased in VLM, whereas in PDM, only A2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable.

Conclusions and Clinical Relevance—Equine muscle contained all Na⁺,K⁺-ATPase mRNA isoforms, but only A2 and B1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na⁺,K⁺-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases.

Abstract

Objective—To determine the effects of short-term IV administration of hydrocortisone or equine growth hormone (eGH) or long-term IM administration of eGH to horses on tissue sensitivity to exogenous insulin.

Animals—5 Standardbreds and 4 Dutch Warmblood horses.

Procedure—The euglycemic-hyperinsulinemic clamp technique was used to examine sensitivity of peripheral tissues to exogenous insulin 24 hours after administration of a single dose of hydrocortisone (0.06 mg/kg), eGH (20 µg/kg), or saline (0.9% NaCl) solution and after long-term administration (11 to 15 days) of eGH to horses. The amounts of metabolized glucose (M) and plasma insulin concentration (I) were determined.

Results—Values for M and the M-to-I ratio were significantly higher 24 hours after administration of a single dose of hydrocortisone than after single-dose administration of eGH or saline solution. After long-term administration of eGH, basal I concentration was increased and the mean M-to-I ratio was 22% lower, compared with values for horses treated with saline solution.

Conclusions and Clinical Relevance—Increases in M and the M-to-I ratio after a single dose of hydrocortisone imply that short-term hydrocortisone treatment increases glucose use by, and insulin sensitivity of, peripheral tissues. Assuming a single dose of hydrocortisone improves sensitivity of peripheral tissues to insulin, it may be an interesting candidate for use in reducing insulin resistance in peripheral tissues of horses with several disease states. In contrast, long-term administration of eGH decreased tissue sensitivity to exogenous insulin associated with hyperinsulinemia. Therefore, increased concentrations of growth hormone may contribute to insulin resistance in horses with various disease states. (Am J Vet Res 2005;66:1907–1913)

Abstract

Objective—To evaluate alterations in skeletal muscle carnitine metabolism during exercise and training by measuring changes in plasma acylcarnitine concentrations in Standardbreds.

Animals—10 Standardbred geldings with a mean ± SD age of 20 ± 2 months and weight of 384 ± 42 kg.

Procedures—In a 32-week longitudinal study, training on a treadmill was divided into 4 phases as follows: phase 1, acclimatization for 4 weeks; phase 2, 18 weeks with alternating endurance and high-intensity exercise training; phase 3, increased training volume and intensity for another 6 weeks; and phase 4, deconditioning for 4 weeks. In phase 3, horses were randomly assigned to 2 groups as follows: control horses (which continued training at the same level as in phase 2) and high-intensity exercise trained horses. At the end of each phase, a standardized exercise test (SET) was performed. Plasma acylcarnitine, fatty acids, and lactic acid and serum β-hydroxybutyric acid (BHBA) concentrations were assessed before and at different time points after each SET.

Results—Plasma lactic acid, total nonesterified fatty acids, 3-hydroxyisobutyric acid, and acetylcarnitine (C2-carnitine) concentrations significantly increased during SETs, whereas serum BHBA, plasma propionylcarnitine (C3-carnitine), and plasma butyryl- and isobutyrylcarnitine (C4-carnitine) concentrations decreased significantly, compared with those before SETs.

Conclusions and Clinical Relevance—Our findings indicated that the plasma acylcarnitine profile in horses likely reflects skeletal muscle carnitine metabolism following exercise, thereby providing a possible practical method to investigate potential disorders in carnitine metabolism in horses with myopathy.

Abstract

Objectives—To determine whether increased glucose metabolism is the potential cause of the decreased plasma glucose curve determined after oral glucose tolerance testing in horses with lower motor neuron degeneration.

Animals—3 horses with signs suggestive of lower motor neuron degeneration, 1 horse with malignant melanoma with multiple metastases, and an obese but otherwise healthy horse.

Procedures—Glucose metabolism was assessed by use of the hyperglycemic clamp and euglycemic hyperinsulinemic clamp techniques.

Results—Mean rate of glucose metabolism of horses with lower motor neuron degeneration was significantly greater (mean, 3.7 times greater than control horses; range, 2.1 to 4.8 times greater) than that reported in 5 healthy control horses (41 ± 13 µmol/kg/min vs 11 ± 4.5 µmol/kg/min, respectively). In addition, one of the affected horses, an 8-year-old warmblood gelding, had a 5.6- times increased sensitivity to exogenously administered insulin, compared with that reported in 5 healthy control horses. Pancreatic insulin secretion was not insufficient in horses with lower motor neuron degeneration. Findings in the 2 diseased control horses were unremarkable.

Conclusions and Clinical Relevance—Increased glucose metabolism in horses with lower motor neuron degeneration may be the cause of the decreased plasma glucose curve detected after oral glucose tolerance testing. This finding could aid in developing supportive treatments with respect to adequate glucose and vitamin E supplementation. (Am J Vet Res 2005;66:271–276)