phosphocreatine and Starvation

Topics: Adenosine Triphosphate; Anatomy, Comparative; Animals; Blood Glucose; Brain Stem; Cachexia; Cats; Cerebral Ventricles; Electric Stimulation; Enteral Nutrition; Feedback; Feeding and Eating Disorders; Feeding Behavior; Globus Pallidus; Goats; Haplorhini; Humans; Hunger; Hypothalamus; Limbic System; Liver; Motor Cortex; Norepinephrine; Obesity; Phosphocreatine; Physiology, Comparative; Rabbits; Rats; Self Stimulation; Sheep; Starvation

Myocellular creatine, which is critically important for normal energy metabolism, increases in rat gastrocnemius muscle after starvation via unknown mechanisms. Creatine (Cr) uptake across plasma membranes is governed by a single, specific transporter (CrTr) that shares 50% amino acid sequence identity with GABA/choline/betaine transporters whose functions are modulated by phosphorylation.. Gastrocnemius muscle was collected from adult male Sprague-Dawley (225-250 g) rats that were randomized to receive normal rat chow and distilled water ad libitum (CTL) or distilled water alone for 4 days (STV). Total Cr, phosphocreatine (PCr), free Cr, and ATP were measured luminometrically. CrTr protein expression and protein serine and tyrosine phosphorylation and mRNA expression were determined using immunoprecipitation and quantitative Western blotting and reverse transcription polymerase chain reaction (RT-PCR) analyses, respectively. Guanidinoacetate methyltransferase (GAMT) activity, guanidinoacetic acid (GAA) content, creatine kinase (CK) activity, and creatinine (Crn) content were assayed luminometrically or spectrophotometrically. Creatine transporter uptake activity was also measured in skeletal muscle membrane vesicles. Data were analyzed by t test.. Total Cr and free Cr increased 26 and 280% in STV (32.3 +/- 1.0 and 12.9 +/- 1.4 vs 25.7 +/- 1.1 and 3.4 +/- 0.9 micromol/g wet wt, mean +/- SEM, respectively, P < 0.01) whereas PCr content decreased 18% (18.6 +/- 0.8 vs 22.8 +/- 0.9 micromol/g wet wt, STV vs CTL P < 0.05). CrTr protein and mRNA expression, ATP, GAA, CK, GAMT, and protein tyrosine phosphorylation of CrTr were not significantly different between the two groups. However, protein serine phosphorylation of CrTr was significantly reduced by 30% (P < 0.05) and creatine uptake activity was significantly increased (P < 0.05) in starved animals.. Increases in myocellular creatine content after starvation are associated with reduced serine phosphorylation of the creatine transporter.

Topics: Animals; Creatine; Creatine Kinase; Gene Expression; Glycine; Guanidinoacetate N-Methyltransferase; Male; Membrane Transport Proteins; Methyltransferases; Muscle, Skeletal; Phosphocreatine; Phosphorylation; Rats; Rats, Wistar; RNA, Messenger; Specific Pathogen-Free Organisms; Starvation

Starvation significantly alters the distribution of body water. To study the effects of starvation on cellular energetics and water distribution in skeletal muscle, a novel 31P magnetic resonance technique (31P MRS) was developed to measure water compartments. After 31P MRS-visible water space markers which distribute in total body water (dimethyl methylphosphonate, DMMP) and extracellular water (phenylphosphonate, PPA) were infused intravenously, 31P MRS spectra were obtained from the gastrocnemius muscle of male virus-free Wistar rats at baseline and after starvation or ad libitum feeding for 4 days. Muscle water spaces were also measured using the chloride method and Nernst's equation. Muscle water contents as determined by drying were equivalent in the two groups. In vivo measurements of changes in DMMP relative to all of the MRS visible phosphates also demonstrated that the total water space was similar in control and starved rats. However, starvation significantly increased the ratio of PPA/DMMP (0.67 +/- 0.05 vs 0.87 +/- 0.04, Control vs Starvation; P < 0.001), and therefore the ratio of extracellular water to total water in the gastrocnemius. Furthermore, because muscle water contents were comparable between the groups, this expansion of the extracellular space was accompanied by contraction of the intracellular compartment in starved animals. Equivalent changes were detected in vitro using the chloride method. Lastly, phosphocreatine/ATP ratios, which measured changes in high-energy phosphate stores, decreased after starvation (4.09 +/- 0.06 vs 3.61 +/- 0.06; P < 0.001) and were inversely related to changes in PPA/DMMP (r = -0.61; P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenosine Triphosphate; Animals; Body Water; Body Weight; Extracellular Space; Hydrogen-Ion Concentration; Magnetic Resonance Imaging; Male; Muscles; Organophosphorus Compounds; Phosphocreatine; Phosphorus; Rats; Rats, Wistar; Starvation

1. The muscle tension and the state of high-energy phosphate metabolism during contraction of the sartorius muscle in frogs (Rana catesbeiana) starved for 1-5 months was studied by in vivo 31P-NMR spectrometry. 2. Muscle tension began to decrease after 2-month starvation compared with the control group and decreased to about one-third of the control value after a 5-month starvation. 3. Muscle contraction induced by electrical stimulation or the use of anaerobic perfusion fluid did not decrease the concentration of creatine phosphate (PCr) or beta-ATP, and only negligibly changed the PCr/Pi ratio from starvation. 4. These results suggest a decrease in creatine kinase activity in the muscle of starved frogs.

Topics: Animals; Body Weight; Electric Stimulation; Energy Metabolism; Magnetic Resonance Spectroscopy; Muscle Contraction; Muscles; Phosphates; Phosphocreatine; Phosphorus Isotopes; Rana catesbeiana; Starvation

The aim of this study was to evaluate whether food intake modulates experimental tumour growth by acute alterations in the energy state and blood flow of the tumour, and if so whether such changes are related to alterations in the enzyme ornithinedecarboxylase (ODC) and DNA synthesis. Inbred mice (C57BL/J) bearing a syngeneic undifferentiated and rapidly growing tumour were used. The tumour levels of high energy phosphates were measured in vivo by 31-P-NMR spectroscopy and biochemically following tissue extraction. DNA synthesis was estimated by measuring the incorporation of bromodeoxy-uridine into tumour DNA. Difluoro-methylornithine (DFMO) was used to inhibit ODC-activity. Tumour blood flow was estimated by a 132Xe local clearance technique. Tumour progression was associated with a significant decrease in tumour tissue high energy phosphates. Acute starvation decreased DNA-synthesis and tumour energy charge as well as its PCr/Pi which were rapidly normalised during subsequent refeeding. These changes were related to similar alterations in tumour blood flow. The inorganic phosphate (Pi) resonance and the resonances in the phosphomonoester (PME) region were considerably increased in tumour tissue. Inhibition of ODC-activity by DFMO decreased DNA-synthesis, which was associated with a secondary increase in tumour high energy phosphates probably due to a lowered energy demand for tumour cell division. The results demonstrate that host undernutrition was translated into retarded tumour growth associated with a decrease in the energy state and blood flow of the tumour. The results have bearing for the evaluation and planning of all treatment protocols with potential influence on food intake in experimental tumour-bearing animals.

Topics: Adenosine Triphosphate; Animals; Eating; Eflornithine; Energy Metabolism; Female; Magnetic Resonance Spectroscopy; Male; Methylcholanthrene; Mice; Mice, Inbred C57BL; Organ Size; Ornithine Decarboxylase; Phosphocreatine; Phosphorus; Phosphorus Isotopes; Sarcoma, Experimental; Starvation

The influence of Ca2+ on myofibrillar proteolysis was evaluated in the isolated extensor digitorum longus muscle incubated in vitro with agents previously shown to increase the intracellular concentration of Ca2+. Myofibrillar proteolysis was evaluated by measuring the release of N tau-methylhistidine, and total proteolysis was evaluated by measuring tyrosine release by incubated muscles after the inhibition of protein synthesis with cycloheximide. Incubated muscles released measurable quantities of N tau-methylhistidine, and muscle contents of the amino acids remained stable over 2 h of incubation. The release of N tau-methylhistidine by incubated muscles was similar to its release by perfused rat muscle in response to brief starvation, indicating the integrity of the incubated muscles. Ca2+ ionophore A23187, dibucaine, procaine, caffeine and elevated K+ concentration increased lactate release by incubated muscles and decreased tissue contents of ATP and phosphocreatine to varying degrees, indicating the metabolic effectiveness of the agents tested. Only A23187 and dibucaine increased total cell Ca2+, and they increased tyrosine release. Caffeine and elevated [K+] increased neither cell Ca2+ nor tyrosine release; however, only A23187 and dibucaine increased tyrosine release significantly. On the other hand, these agents were without effect on myofibrillar proteolysis as assessed by N tau-methylhistidine release by incubated muscles and changes in tissue contents of the amino acid. In fact, some of the agents tested tended to decrease myofibrillar proteolysis slightly. These results indicate that acute elevation of intracellular Ca2+ is associated with increased breakdown of non-myofibrillar but not myofibrillar proteins. Because of this, the role of elevated Ca2+ in muscle atrophy in certain pathological states is questioned. The data also indicate that the breakdown of myofibrillar and non-myofibrillar proteins in muscle is regulated independently and by different pathways, a conclusion reached in previous studies with perfused rat muscle.

Topics: 2,4-Dinitrophenol; Adenosine Triphosphate; Animals; Calcium; Dinitrophenols; Lactates; Lactic Acid; Male; Methylhistidines; Muscle Proteins; Phosphocreatine; Rats; Rats, Inbred Strains; Starvation; Tyrosine

A method was developed for the hemoglobin-free perfusion of the rabbit psoas muscle in situ. This muscle consists almost exclusively of fast-twitch (type IIb) glycolytic fibres and was therefore used as a model of a homogeneous muscle of the glycolytic, metabolic type.

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Glucose; Glycogen; Glycolysis; In Vitro Techniques; Insulin; Kinetics; Male; Muscles; Perfusion; Phosphocreatine; Rabbits; Starvation

To determine if vanadate has insulin-like actions in skeletal muscle, we measured its effects on glucose and protein metabolism in epitrochlearis muscles of rats. Compared with insulin, vanadate increased glucose uptake, glycogen synthesis and glycolysis to a lesser degree, but caused a greater stimulation of lactate and glucose oxidation. Unlike insulin, vanadate did not change either protein synthesis or degradation. These different metabolic responses could be related to the different pattern of insulin-receptor phosphorylation caused by insulin and vanadate.

Topics: Adenine Nucleotides; Animals; Glucose; In Vitro Techniques; Insulin; Lactates; Lactic Acid; Male; Muscle Proteins; Muscles; Phosphocreatine; Rats; Rats, Inbred Strains; Starvation; Vanadates; Vanadium

The effect of ischaemia on the concentration of active pyruvate dehydrogenase complex has been investigated in glucose perfused hearts of normal rats fed a normal diet or a high fat diet or starved for 48 h; and in hearts from alloxan-diabetic rats. Global ischaemia induced by low flow (approx. 1 ml/min) lowered the concentration of active complex under most of the experimental conditions employed. Parallel studies showed that anoxia and K+ arrest of the heart had effects similar to that of ischaemia and suggested that hypoxia and decreased mechanical activity of the heart may be responsible for effects of low flow ischaemia. Evidence is reviewed that the effects of low flow ischaemia, K+ arrest and anoxia may be mediated through activation of pyruvate dehydrogenase kinase by increased reduction of mitochondrial NAD+. In hearts of normal rats on a normal diet, global ischaemia induced by zero flow and regional ischaemia induced by coronary artery ligation increased the concentration of active complex. Evidence is given that this may result from a combination of anoxia and acidosis. In aerobic perfusions at 60 mmHg, concentrations of active complex were ranked in the order: normal diet greater than high fat diet greater than 48 h starved greater than alloxan diabetic. This order was maintained when the concentration of active complex was increased by perfusion at 120 mmHg or lowered by global ischaemia induced by zero flow.

Topics: Adenosine Triphosphate; Animals; Coronary Disease; Diabetes Mellitus, Experimental; Dietary Fats; Heart Arrest; Lactates; Lactic Acid; Male; Myocardium; Oxygen Consumption; Perfusion; Phosphocreatine; Pyruvate Dehydrogenase Complex; Rats; Rats, Inbred Strains; Starvation

The rate of protein synthesis was measured in muscles of the perfused rat hemicorpus, and values were compared with rates obtained in whole animals. In gastrocnemius muscle of fed rats the rate of synthesis measured in the hemicorpus was the same as that in the whole animal. However, in plantaris, quadriceps and soleus muscles rates were higher in the hemicorpus than those in vivo. In the hemicorpus, starvation for 1 day decreased the rate of protein synthesis in gastrocnemius and plantaris muscles, in parallel with decreases in the RNA content, but the soleus remained unaffected. Similar effects of starvation were observed in vivo, so that the relationships between rates in vivo and in the hemicorpus were the same as those in fed rats. Proteins of quadriceps and plantaris muscles were separated into sarcoplasmic and myofibrillar fractions. The rate of synthesis in the sarcoplasmic fraction of the hemicorpus from fed rats was similar to that in vivo, but synthesis in the myofibrillar fraction was greater. In the plantaris of starved rats the rates of synthesis in both fractions were lower, but the relationships between rates measured in vivo and in the perfused hemicorpus were similar to those seen in fed rats. The addition of insulin to the perfusate of the hemicorpus prepared from 1-day-starved animals increased the rates of protein synthesis per unit of RNA in gastrocnemius and plantaris muscles to values above those seen in fed animals when measured in vivo or in the hemicorpus. Insulin had no effect on the soleus. Overall, the rates of protein synthesis in the hemicorpus differed from those in vivo. However, the effect of starvation when measured in the whole animal was very similar to that measured in the isolated rat hemicorpus when insulin was omitted from the perfusate.

Topics: Adenine Nucleotides; Animals; Food; Kinetics; Male; Muscle Proteins; Muscles; Myofibrils; Perfusion; Phosphocreatine; Rats; Rats, Inbred Strains; RNA; Sarcoplasmic Reticulum; Starvation; Tyrosine

Topics: Adenine Nucleotides; Animals; Blood-Brain Barrier; Brain; Diabetes Mellitus; Diabetes Mellitus, Experimental; Diabetic Coma; Diabetic Ketoacidosis; Energy Metabolism; Glucose; Humans; Ketone Bodies; Oxygen Consumption; Phosphocreatine; Rats; Starvation

Topics: Animals; Aurintricarboxylic Acid; Creatine Kinase; Guanine Nucleotides; Guanosine Triphosphate; Kinetics; Magnesium; Methionine; Muscles; Myocardium; Organ Specificity; Peptide Initiation Factors; Phosphocreatine; Rats; RNA, Transfer; Spermidine; Spermine; Starvation

1. The effects of starvation and diabetes on brain fuel metabolism were examined by measuring arteriovenous differences for glucose, lactate, acetoacetate and 3-hydroxybutyrate across the brains of anaesthetized fed, starved and diabetic rats. 2. In fed animals glucose represented the sole oxidative fuel of the brain. 3. After 48h of starvation, ketone-body concentrations were about 2mm and ketone-body uptake accounted for 25% of the calculated O(2) consumption: the arteriovenous difference for glucose was not diminished, but lactate release was increased, suggesting inhibition of pyruvate oxidation. 4. In severe diabetic ketosis, induced by either streptozotocin or phlorrhizin (total blood ketone bodies >7mm), the uptake of ketone bodies was further increased and accounted for 45% of the brain's oxidative metabolism, and the arteriovenous difference for glucose was decreased by one-third. The arteriovenous difference for lactate was increased significantly in the phlorrhizin-treated rats. 5. Infusion of 3-hydroxybutyrate into starved rats caused marked increases in the arteriovenous differences for lactate and both ketone bodies. 6. To study the mechanisms of these changes, steady-state concentrations of intermediates and co-factors of the glycolytic pathway were determined in freeze-blown brain. 7. Starved rats had increased concentrations of acetyl-CoA. 8. Rats with diabetic ketosis had increased concentrations of fructose 6-phosphate and decreased concentrations of fructose 1,6-diphosphate, indicating an inhibition of phosphofructokinase. 9. The concentrations of acetyl-CoA, glycogen and citrate, a potent inhibitor of phosphofructokinase, were increased in the streptozotocin-treated rats. 10. The data suggest that cerebral glucose uptake is decreased in diabetic ketoacidosis owing to inhibition of phosphofructokinase as a result of the increase in brain citrate. 11. The inhibition of brain pyruvate oxidation in starvation and diabetes can be related to the accelerated rate of ketone-body metabolism; however, we found no correlation between the decrease in glucose uptake in the diabetic state and the arteriovenous difference for ketone bodies. 12. The data also suggest that the rates of acetoacetate and 3-hydroxybutyrate utilization by brain are governed by their concentrations in plasma. 13. The finding of very low concentrations of acetoacetate and 3-hydroxybutyrate in brain compared with plasma suggests that diffusion across the blood-brain barrier

Topics: Acetoacetates; Acetyl Coenzyme A; Adenine Nucleotides; Animals; Biological Transport; Blood Glucose; Blood-Brain Barrier; Brain; Citrates; Diabetes Mellitus; Female; Glucose; Glycogen; Hydroxybutyrates; Ketone Bodies; Lactates; Pentobarbital; Phlorhizin; Phosphocreatine; Rats; Spectrometry, Fluorescence; Starvation; Streptozocin; Time Factors

Topics: Adenosine Triphosphate; Animals; Bone and Bones; Brain Chemistry; Drug Stability; Liver; Methanol; Methods; Perchlorates; Phosphates; Phosphocreatine; Rats; Starvation

Topics: Adenine Nucleotides; Animals; Calcium; Dogs; Energy Metabolism; Glucose; Heart Failure; Hypoxia; In Vitro Techniques; Male; Mitochondria, Muscle; Myocardial Contraction; Myocardium; Phosphocreatine; Rats; Sarcoplasmic Reticulum; Starvation

Topics: Adenosine Triphosphate; Animals; Chlorothiazide; Diabetes Mellitus, Experimental; Glucose; Glycogen; Hexosephosphates; Insulin; Ischemia; Kidney; Lactates; Male; Organomercury Compounds; Phlorhizin; Phosphocreatine; Rats; Starvation