Insulin action on whole body glucose utilization and on muscle glucose transporter translocation in mice

Murine models of insulin resistance and diabetes are versatile and have been used to investigate genetic and metabolic disorders. However, the principal assays to assess insulin action, i.e., the euglycemic-hyperinsulinemic clamp and subcellular distribution of glucose transporters, have not been implemented in this species. Here we describe procedures which allow these methods to be adapted to mice. When normal C57bl/6j mice were infused with graded doses of insulin (1, 3, 10 or 30 mU/kg/min) during a euglycemic-hyerinsulinemic clamp, the glucose infusion rate necessary to maintain euglycemia increased in a dose-dependent manner (7.4 +/- 1.7, 13.1 +/- 3.6, 24.1 +/- 2.3 or 34.8 +/- 7.5 mg/kg/min), respectively. Hindlimb muscles were isolated and samples of 2-3 g were subjected to subcellular fractionation finalizing on 25%, 30% and 35% sucrose gradients. Fraction F25 (plasma membranes) was enriched in alpha 2 Na+/K(+)-ATPase and GLUT1 glucose transporters, whereas fraction F35 (intracellular membranes) was enriched in Ca(2+)-ATPase and GLUT4 glucose transporters. Following insulin treatment, GLUT4 increased in F25 and decreased in F35. Insulin treatment had no effect on GLUT1 in F25. However, unlike in rat skeletal muscle, GLUT1 was detectable in F35 and its content decreased in this fraction following insulin treatment. The results demonstrate that whole-body glucose utilization can be assessed in mice using euglycemic-hyperinsulinemic clamps and demonstrate how subcellular fractionation procedures can be applied to murine muscle. Murine muscle GLUT4 translocates from an intracellular storage site to the plasma membrane in response to insulin.     

Acute and chronic effects of leptin on glucose utilization in lean mice

Experiments described here show that in vivo glucose uptake is impaired in mice given 30 micrograms leptin by intraperitoneal injection 2 hours before an oral glucose tolerance test (GTT). When mice were infused for 7 days with 10 micrograms/day leptin, the 4-fold increase in circulating leptin caused a transient hypophagia, a sustained weight loss and significantly inhibited insulin release in response to an oral GTT. Adipocytes from these mice were not insulin responsive whereas insulin-stimulated muscle and liver glycogen synthesis were increased. In contrast, leptin added to 2 hour in vitro incubations had an insulin-like effect on muscle glucose utilization and augmented insulin stimulation of adipocyte lipid synthesis. Thus, normal mice treated chronically with leptin develop tissue specific changes in insulin sensitivity and compensate for inhibition of glucose-stimulated insulin release. The contrasting response to acute leptin exposure suggests these changes are not a direct effect of the protein.     

Decreased cerebral glucose utilization in rats during the ebb phase of thermal injury

OBJECTIVE: Thermal injury is associated with the development of encephalopathy. The mechanism(s) for the development of this condition have not been established. In the present study, the effects of thermal injury were determined on rat brain glucose utilization (Rg), using 2-[18F]fluoro-2-deoxy-D-glucose (18FDG). DESIGN: Four types of studies were performed. In one group of rats, the effect of thermal injury on total rat brain glucose utilization (Rg) was determined at 6 hours, 24 hours, and 3 weeks after injury. The brains of thermally injured rats were also assayed for hexokinase and glucose-6-phosphatase activities, since these enzyme activities are responsible for the phosphorylation and dephosphorylation of the 18FDG. We also measured total body oxygen consumption in the thermally injured rats. We wanted to compare the changes produced by thermal injury on rat brain glucose utilization (Rg) with the effects produced by compounds known to modify energy metabolism and/or rat brain glucose utilization (Rg). For that reason, in a second group of rats, an inflammatory state was produced by lipopolysaccharide injection, and rat brain glucose utilization (Rg) was determined. In the third group of rats, overall metabolism in rats was reduced by pentobarbital injection, followed by hypothermia, and rat brain glucose utilization (Rg) was determined. In the fourth group of rats, overall metabolism in rats was stimulated by 2,4-dinitrophenol injection, and rat brain glucose utilization (Rg) was determined. MATERIALS AND METHODS: Glucose utilization (Rg) by the brains of these treated rats was determined using 18FDG. Oxygen consumption in vivo, as well as glucose-6-phosphatase and hexokinase activity in vitro, were measured by standard procedures. MEASUREMENTS AND MAIN RESULTS: Glucose utilization (Rg) by rat brain was significantly reduced (p < 0.01) at 6 and 24 hours after injury, but returned to normal values 3 weeks after injury. These reductions were associated with decreases in rat brain hexokinase activity, increases in rat brain glucose-6-phosphatase activity, and decreased oxygen consumption by rats in vivo. Pentobarbital injection followed by hypothermia reduced rat brain glucose utilization (Rg) (p < 0.01), while 2,4-dinitrophenol treatment elevated rat brain glucose utilization (Rg) (p < 0.01). In contrast, LPS treatment had no effect on rat brain glucose utilization (Rg). CONCLUSIONS: These data indicate that thermal injury decreases glucose utilization (Rg) in rat brain during the hypometabolic phase. This effect can be explained, at least in part, by alterations in hexokinase and glucose-6-phosphatase activities, as well as reductions in oxygen consumption. Thus, the changes in brain glucose utilization (Rg) appear to be associated with the ebb phase of the thermal injury. The present results observed in burned rats may provide evidence to explain the encephalopathy observed in burned patients.     

Intracerebroventricular administration of leptin markedly enhances insulin sensitivity and systemic glucose utilization in conscious rats

This study examines the acute, subacute (overnight), and chronic (7-day) effects of intracerebroventricular (i.c.v.) administration of r-metMuLeptin on insulin sensitivity and systemic glucose turnover in conscious unrestrained rats (body weight, 250 to 300 g). Under postabsorptive conditions, acute i.c.v. leptin ([AL] 10 microg bolus) did not affect tracer (3-(3)H-glucose)-determined glucose production (GP) and utilization (GU) rates during the 2-hour hyperinsulinemic (2 mU x kg(-1) x min(-1)) euglycemic clamp. Chronic i.c.v. leptin ([CL] 10 microg/d for 7 days) administered by osmotic pumps markedly reduced the daily food consumption (P < .05), body weight (P < .05), and postabsorptive basal plasma glucose level (P < .01). During the glucose clamp, GP was markedly suppressed (55%) with CL (P < .001 v vehicle and pair-fed control groups). The insulin-induced increment in GU was significantly greater with CL (23.3 +/- 1.8 mg(-1) x kg(-1) x min(-1)) than with vehicle (16.9 +/- 0.2) and pair-feeding (17.1 +/- 0.6, both P < .001). Subacute i.c.v. leptin ([SL] 10 microg bolus) moderately but insignificantly decreased overnight food consumption (-18%) and body weight (-2.5 +/- 1.5 g). The glucose infusion rate during the final 60 minutes of the glucose clamp was 43% greater than for the vehicle group (P < .0001). SL also significantly increased GU (P < .005) and suppressed GP (P < .05) during the glucose clamp. Thus, we conclude that i.c.v. administered leptin has strong actions on the central nervous system that result in significant increases in insulin sensitivity and systemic GU, and these effects are achieved as early as overnight after leptin administration.     

d-Amphetamine attenuates decreased cerebral glucose utilization after unilateral sensorimotor cortex contusion in rats

Unilateral contusion injury to the sensorimotor cortex causes, among other symptoms, a transient contralateral hindlimb hemiparesis in rats. A single i.p. 2 mg/kg dose of d-amphetamine (d-AMPH) 24 h after injury accelerates spontaneous recovery from this particular deficit. The mechanism(s) of spontaneous and d-AMPH enhanced recovery are unknown but alleviation of a neuronal depression has been proposed. This quantitative CMRglu study was designed to determine effects of cortical contusion injury and d-AMPH on CMRglu in cortical and subcortical structures. At 2 days after injury, CMRglu was significantly reduced compared to sham-operated controls only in structures ipsilateral to contusion. Affected structures included the caudate putamen, medial geniculate nucleus, lateral geniculate nucleus and the parietal cortex immediately posterior to injury. By 6 days post-contusion, the hypometabolism partially reversed in all structures. A single low dose of d-AMPH significantly alleviated the post-traumatic CMRglu reduction at 2 days after injury. Importantly, while this alleviation was not significant for any single structure, the main effect of treatment was highly significant. d-AMPH increased CMRglu at 2 days post-injury by 18-33% compared to contused/saline-treated rats. These results suggest that alleviation of neuronal metabolic depression may contribute to spontaneous and d-AMPH enhanced recovery.     

Suppression of glycolysis is associated with an increase in glucose cycling in hepatocytes from diabetic rats

Rates of cycling between glucose and glucose 6-phosphate and between glucose and pyruvate, and the effects of these cycles on glucose metabolism, were compared in hepatocytes isolated from fasted normal or streptozotocin-induced diabetic rats. In diabetic hepatocytes the rate of glucose phosphorylation was 30% lower than that in normal hepatocytes, and there was a doubling of the rate of glucose/glucose 6-phosphate cycling. In addition, the rate of glycolysis was 60% lower in diabetic hepatocytes. This inhibition of glycolysis and stimulation of glucose/glucose 6-phosphate cycling appeared to be a consequence of the elevated rates of endogenous fatty acid oxidation observed in diabetic hepatocytes. The proportion of glycolytically derived pyruvate that was recycled to glucose was more than doubled in hepatocytes from diabetic rats compared with normal animals. This increase also appeared to be linked to the high rates of endogenous fatty acid oxidation in diabetic cells. As a consequence of the increased rates of both these cycles, 85% of all glucose molecules taken up by diabetic hepatocytes were recycled to glucose, compared with only 50% in normal hepatocytes. Glucose cycling is therefore likely to make a substantial contribution to the hyperglycemia of diabetes.     

Medial forebrain bundle lesions: Specific loss of feeding to decreased glucose utilization in rats.

Found that bilateral destruction of the medial forebrain bundle in female Sherman rats eliminated feeding to decreased intracellular glucose utilization (glucoprivation). The deficit was specific. Feeding was enhanced by dietary dilution and reduced by dietary concentration. More was eaten in the cold and less in the heat. The Ss were not differentially sensitive to quinine adulteration. They returned to normal body weight following regimens of gavage or of restricted feeding. Moreover, they did not differ from normal Ss in drinking to various thirst stimuli. It is suggested that the glucoprivic mechanism makes but a minor contribution to the initiation of spontaneous feeding. (39 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effects of the self-administration of ethanol and ethanol/sucrose on rates of local cerebral glucose utilization in rats

In a previous study, the voluntary ingestion of ethanol by rats was found to be associated with a discrete pattern of changes in functional activity that included the nucleus accumbens, medial prefrontal cortex, basolateral and central nuclei of the amygdala, as well as the ventral midbrain. Rats in this study, however, consumed a combination of ethanol in a sucrose vehicle. The purpose of the present experiment was to characterize the role of sucrose in determining the effects of orally self-administered ethanol using the quantitative autoradiographic 2-[14C]deoxyglucose (2DG) method for measurement of rates of local cerebral glucose utilization. A modified sucrose-substitution procedure was employed to train three groups of Wistar rats to self-administer either water, 10% ethanol (10E), or a 10% ethanol/2% sucrose solution (10E/2S) in daily sessions. An additional group of rats was trained using a modified acclimation procedure (home cage) in order to determine if any exposure to sucrose would alter rates of glucose utilization. Once stable rates of consumption were established, the 2DG method was applied immediately following completion of the final test session. Rats received a dose of ethanol equivalent to 0.5 g kg-1 on the day of the procedure or a comparable volume of water. Rates of energy metabolism were significantly increased in all three groups of rats that consumed ethanol (10E/2S, 10E, and home cage), as compared to rates in rats that consumed water. The areas of significant change included the rostral pole and posterior shell of the nucleus accumbens, medial prefrontal cortex, the basolateral and central nuclei of the amygdala, the ventral tegmental area, and the substantia nigra pars compacta. Thus, the pattern of changes in functional brain activity that accompanies voluntary ingestion of ethanol is independent of the vehicle in which the ethanol is presented or the procedures used to initiate consumption. Furthermore, these data demonstrate that it is the simultaneous activation of an interrelated network of limbic brain regions that serves as the substrate of the effects of ethanol self-administration.     

Cerebral utilization of glucose, ketone bodies and oxygen in starving infant rats and the effect of intrauterine growth retardation

Cerebral arteriovenous differences of acetoacetate, D-beta-hydroxybutyrate, glucose, lactate and oxygen and brain DNA content was measured at 20 days of age in intrauterine growth retarded (IUGR) rats and normal littermates after 48 and 72 h of starvation. Cerebral blood flow (CBF) was measured with labeled microspheres in other comparable groups of IUGR and control rats. CBF was similar in IUGR and normal littermates (0.57+/-0.09 and 0.58+/-0.10 ml/min respectively). After 48 h of starvation, arterial glucose was significantly lower in IUGR than control animals but the arterial concentrations of ketone bodies were similar. After 48 h of starvation, cerebral arteriovenous difference of beta-hydroxybutyrate was significantly higher in control than IUGR rats also when expressed per mg brain DNA as was the fractional uptake of D-beta-hydroxybutyrate. After 72 h of starvation, arterial concentrations of ketone bodies were significantly lower in IUGR rats than controls but the fractional uptake of D-beta-hydroxybutyrate was increased compared to IUGR rats starved for 48 h. The average percentage of calculated total substrate uptake (mumol/min) accounted for by ketone bodies increased in control animals from 31.1% after 48 h of starvation to 41.0% after 72 h of starvation. In IUGR rats these percentage values were 26.5 and 25.7 respectively. After 72 h of starvation the fraction of total cerebral uptake of substrates accounted for by ketone bodies was significantly higher in control that IUGR rats. As total cerebral uptake of substrates was similar between IUGR and control animals it is concluded that IUGR rats are more dependent on glucose as a substrate for the brain during starvation.     

Fuel utilization and glucose hyperalimentation after liver resection

Clinical studies and experiments in rats were carried out to elucidate changes in fuel utilization after hepatectomy. In addition, the effect of glucose hyperalimentation on energy metabolism in the liver remnant was studied. Respiratory quotient (RQ) and substrate oxidation rate for fat and glucose were evaluated by indirect calorimetry in eight patients who had undergone liver resection. Patients had a reduced nonprotein RQ of approximately 0.85 and a reduced ratio of glucose to fat oxidation of approximately 2.0 on the 1st and 2nd postoperative days. After 80% hepatectomy, rats received either 30 kcal.kg-1.day-1 (group 1) or 200 kcal.kg-1.day-1 (group 2) of glucose for 48 h. In both rat groups, hepatic mitochondrial ATP synthesis 12 and 24 h after hepatectomy was accelerated when palmitic acid was used as the substrate and suppressed when pyruvate was used compared with sham-operated groups. This suggests that the energy substrate of the remnant liver was principally fatty acids rather than glucose, which seems to occur also in humans. Hepatic energy charge was within normal limits in group 1 (0.862 +/- 0.008) but decreased significantly in group 2 (0.818 +/- 0.006, p < 0.01) 12 h after hepatectomy. An abundance of glucose in the early postoperative period therefore caused a hepatic energy derangement by suppressing endogenous fat oxidation. This suppression was corroborated by the findings of lower immunoreactive glucagon and nonesterified fatty acid concentration in group 2. Therefore, glucose hyperalimentation in the early postoperative period after liver resection is not recommended.     

Control of glucose utilization in working perfused rat heart

Metabolic control analyses of glucose utilization were performed for four groups of working rat hearts perfused with Krebs-Henseleit buffer containing 10 mM glucose only, or with the addition of 4 mM D-beta-hydroxybutyrate/1 mM acetoacetate, 100 nM insulin (0.05 unit/ml), or both. Net glycogen breakdown occurred in the glucose group only and was converted to net glycogen synthesis in the presence of all additions. The flux of [2-3H]glucose through P-glucoisomerase (EC 5.3.1.9) was reduced with ketones, elevated with insulin, and unchanged with the combination. Net glycolytic flux was reduced in the presence of ketones and the combination. The flux control coefficients were determined for the portion of the pathway involving glucose transport to the branches of glycogen synthesis and glycolysis. Major control was divided between the glucose transporter and hexokinase (EC 2.7.1.1) in the glucose group. The distribution of the control was slightly shifted to hexokinase with ketones, and control at the glucose transport step was abolished in the presence of insulin. Analysis of the pathway from 3-P-glycerate to pyruvate determined that the major control was shared by enolase (EC 4.2.1.1) and pyruvate kinase (EC 2.7.1.40) in the glucose group. Addition of ketones, insulin, or the combination shifted the control to P-glycerate mutase (EC 5.4.2.1) and pyruvate kinase. These results illustrate that the control of the metabolic flux in glucose metabolism of rat heart is not exerted by a single enzyme but variably distributed among enzymes depending upon substrate availability, hormonal stimulation, or other changes of conditions.     

Influence of diet pellet hardness and particle size on food utilization by mice, rats and hamsters

Increasing hardness of diet pellets reduced food wastage by each species. Also, less wastage occurred when pellets made from finely ground materials were given, an effect that was not related to hardness. The hardest diet reduced growth of the mice by reducing true food consumption and a poorer food conversion efficiency (true food consumption/growth) was obtained. Apparent food consumption increased with the softness of the diet and food utilization (apparent food consumption/growth) of the softest diets was less efficient than those of the others. Grinding of the raw materials prior to pelleting had no effect on food conversion, but food utilization was less efficient because of the greater wastage of pellets from coarsely ground materials and consequent apparent food comsumption.     

Leptin increases glucose transport and utilization in skeletal muscle in vitro

1. The present study examines the effect of leptin on glucose transport and metabolism in incubated soleus muscle from male lean albino rats. 2. Insulin (100 microU/ml) increased glucose uptake by twofold while the leptin group (100 nmol/l) reached 75% of the insulin response after 1 hr of incubation. However, leptin did not potentiate the insulin effect on glucose uptake in soleus muscle. 3. Leptin elicited a significant increase (27.7%) in total lactate production, accompanied by a three-fold increment in glycogen synthesis from [U-14C]D-glucose. 4. Insulin raised glycogen synthesis by sixfold. The leptin plus insulin group increased glycogen synthesis by eightfold, which is equivalent to the sum of the separated leptin and insulin groups. 5. Leptin per se exerts an insulin-like effect stimulating glucose uptake, glycogen synthesis, and lactate formation and also seems to potentiate the effect of insulin on glucose incorporation into glycogen in incubated soleus muscle.     

Trifluoperazine inhibition of insulin-induced increase in skeletal muscle glucose uptake

Transplacental passage of fluorides was studied in 25 randomly selected neonates. Blood samples collected simultaneously from the mother and the umbilical cord showed that average fluoride concentration in the cord blood was 60% of that in mother's blood. When concentration in the mother's blood exceeded 0.4 ppm, the placenta acted as a selective barrier.     

Local cerebral blood flow, local cerebral glucose utilization, and flow-metabolism coupling during sevoflurane versus isoflurane anesthesia in rats

Antepartum plasma hepatitis C virus (HCV) RNA was quantified in 155 mothers coinfected with HCV and human immunodeficiency virus type 1 (HIV-1), and HCV RNA was serially assessed in their infants. Of 155 singleton infants born to HCV antibody-positive mothers, 13 (8.4%) were HCV infected. The risk of HCV infection was 3.2-fold greater in HIV-1-infected infants compared with HIV-1-uninfected infants (17.1% of 41 vs. 5.4% of 112, P = .04). The median concentration of plasma HCV RNA was higher among the 13 mothers with HCV-infected infants (2.0 x 10(6) copies/mL) than among the 142 mothers with HCV-negative infants (3.5 x 10(5) copies/mL; P < .001), and there were no instances of HCV transmission from 40 mothers with HCV RNA concentrations of < 10(5) copies/mL. Women dually infected with HIV-1 and HCV but with little or no detectable HCV RNA should be reassured that the risk of perinatal transmission of HCV is exceedingly low.     

Role of platelet-activating factor in glucose uptake and utilization of different tissues

The particle transport characteristics of two ventilation configurations commonly used in hospital operating rooms (ORs), cross-flow and impinging-flow ventilation, were investigated. The computational fluid dynamics software FLUENT was used to simulate turbulent airflow with mixed convection in a three-dimensional, rectangular OR. Two OR personnel, a patient, OR spotlights, an anesthetics cart, and an operating table were represented in the room. Heat loads from the personnel, patient, and lights affected the airflow through buoyancy. Particles produced at the operation site with various sizes and initial conditions were tracked through the room. A stochastic model was used to include the random effects of turbulence on particle trajectories. Simulation results show that heat loads from the personnel, patient, and OR spotlights had an important effect on the airflow through natural convection. Particle trajectories were influenced greatly by the flow field structure, particle launch position, and turbulence in the flow, and somewhat by particle size. However, particle paths were insensitive to the launch velocity. Virtually identical trajectories were obtained for particles with launch velocities ranging from 0 to 1 m/sec in magnitude. Changes in ventilation configuration dramatically affected particle transport. The cross-flow ventilation configuration performed better, based on the criteria of removing particles from the breathing zone of room occupants. Proper flow field design and contaminant source placement can be used to control particle transport. Numerical simulations allow quick and inexpensive comparisons between room designs and provide details about airflow and contaminant transport.     

Cerebral glucose utilization is reduced in second test session

Cerebral glucose utilization was higher during the first positron emission tomography (PET) session than during the second session, as assayed using the PET [18F]fluorodeoxyglucose method in male human volunteers. This difference was due largely to data from subjects with low-trait anxiety, since subjects with high anxiety showed similar metabolism in both PET sessions. High-anxiety subjects showed greater right/ left ratios of cerebral metabolism than low-anxiety subjects, particularly during the second PET session. These findings suggest that the level of anxiety may be an important variable to consider in PET studies using multiple sessions.