Abundance, localization, and insulin-induced translocation of glucose transporters in red and white muscle

1992 ◽  
Vol 263 (2) ◽  
pp. C443-C452 ◽  
Author(s):  
A. Marette ◽  
J. M. Richardson ◽  
T. Ramlal ◽  
T. W. Balon ◽  
M. Vranic ◽  
...  
Keyword(s):  
Muscle Cell ◽  
Plasma Membranes ◽  
White Muscle ◽  
Glucose Transporter ◽  
Glucose Transporters ◽  
Muscle Spindles ◽  
Molar Ratio ◽  
Type I ◽  
Slow Twitch ◽  
Fast Twitch

D-Glucose protectable cytochalasin B (CB) binding to subcellular membrane fractions was used to determine glucose transporter number in red (quadriceps-gastrocnemius-soleus) and white (quadriceps-gastrocnemius) rat muscle. CB binding was twofold higher in isolated plasma membranes of red than of white muscle. In contrast, the number of transporters in an isolated insulin-sensitive intracellular membrane organelle was similar in the two muscle groups. Immunoblotting and immunofluorescence microscopy with anti-GLUT4 and anti-GLUT1 antibodies indicated that both GLUT1 and GLUT4 transporter isoforms account for the higher abundance of CB binding sites in plasma membranes of red than of white muscle. Immunofluorescence localized GLUT4 to both the surface and the interior of the muscle cell and demonstrated that type I (slow twitch oxidative) and type IIa (fast twitch oxidative-glycolytic) fibers are enriched in GLUT4 protein compared with type IIb (fast twitch glycolytic) fibers. In contrast, GLUT1 reactivity was restricted to the surface of the muscle cell and was also highly enriched in the perineurial sheaths of peripheral nerves and the capsules of muscle spindles present in both red and white muscles. Insulin caused a twofold increase in CB binding in isolated plasma membranes of red or white muscles with a corresponding 40-50% decrease in CB binding in isolated intracellular membranes. These changes in CB binding were paralleled by similar changes in the membrane distribution of the GLUT4 glucose transporter isoform and in glucose transport activity measured after insulin perfusion of hindquarter muscles. In contrast, insulin did not change the distribution of either GLUT1 glucose transporters or Na(+)-K(+)-ATPase alpha 1-subunits. The molar ratio of GLUT4 to GLUT1 in red and white muscle plasma membranes was found to be 4:1 in the basal state and 7:1 in the insulin-stimulated state. These results indicate that red muscle contains a higher amount of GLUT1 and GLUT4 transporters at the plasma membrane than white muscle in the basal and insulin-stimulated states but that GLUT4 translocation does not differ between muscle types. In addition, GLUT4 expression correlates with the metabolic nature (oxidative vs. glycolytic) of skeletal muscle fibers, rather than with their contractile properties (slow twitch vs. fast twitch).

scholarly journals Cycloheximide decreases glucose transporters in rat adipocyte plasma membranes without affecting insulin-stimulated glucose transport

1988 ◽  
Vol 251 (2) ◽  
pp. 491-497 ◽  
Author(s):  
S Matthaei ◽  
J M Olefsky ◽  
E Karnieli
Keyword(s):  
Protein Synthesis ◽  
Glucose Transport ◽  
Plasma Membranes ◽  
Glucose Transporter ◽  
Cytochalasin B ◽  
Glucose Transporters ◽  
Electrophoretic Analysis ◽  
Adipocyte Plasma Membranes ◽  
Inhibitor Of Protein Synthesis ◽  
Adipose Cells

This study examines the relationship between insulin-stimulated glucose transport and insulin-induced translocation of glucose transporters in isolated rat adipocytes. Adipose cells were incubated with or without cycloheximide, a potent inhibitor of protein synthesis, for 60 min and then for an additional 30 min with or without insulin. After the incubation we measured 3-O-methylglucose transport in the adipose cells, and subcellular membrane fractions were prepared. The numbers of glucose transporters in the various membrane fractions were determined by the cytochalasin B binding assay. Basal and insulin-stimulated 3-O-methylglucose uptakes were not affected by cycloheximide. Furthermore, cycloheximide affected neither Vmax. nor Km of insulin-stimulated 3-O-methylglucose transport. In contrast, the number of glucose transporters in plasma membranes derived from cells preincubated with cycloheximide and insulin was markedly decreased compared with those from cells incubated with insulin alone (10.5 +/- 0.8 and 22.2 +/- 1.8 pmol/mg of protein respectively; P less than 0.005). The number of glucose transporters in cells incubated with cycloheximide alone was not significantly different compared with control cells. SDS/polyacrylamide-gel-electrophoretic analysis of [3H]cytochalasin-B-photolabelled plasma-membrane fractions revealed that cycloheximide decreases the amount of labelled glucose transporters in insulin-stimulated membranes. However, the apparent molecular mass of the protein was not changed by cycloheximide treatment. The effect of cycloheximide on the two-dimensional electrophoretic profile of the glucose transporter in insulin-stimulated low-density microsomal membranes revealed a decrease in the pI-6.4 glucose-transporter isoform, whereas the insulin-translocatable isoform (pI 5.6) was decreased. Thus the observed discrepancy between insulin-stimulated glucose transport and insulin-induced translocation of glucose transporters strongly suggests that a still unknown protein-synthesis-dependent mechanism is involved in insulin activation of glucose transport.

Stimulation of glucose transport by thyroid hormone in 3T3-L1 adipocytes: increased abundance of GLUT1 and GLUT4 glucose transporter proteins

2000 ◽  
Vol 164 (2) ◽  
pp. 187-195 ◽  
Author(s):  
R Romero ◽  
B Casanova ◽  
N Pulido ◽  
AI Suarez ◽  
E Rodriguez ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Glucose Uptake ◽  
Glucose Transport ◽  
Plasma Membranes ◽  
Glucose Transporter ◽  
Fold Increase ◽  
Glucose Transporters ◽  
Insulin Binding ◽  
Total Protein Content ◽  
Glucose Transporter Proteins

In 3T3-L1 adipocytes we have examined the effect of tri-iodothyronine (T(3)) on glucose transport, total protein content and subcellular distribution of GLUT1 and GLUT4 glucose transporters. Cells incubated in T(3)-depleted serum were used as controls. Cells treated with T(3) (50 nM) for three days had a 3.6-fold increase in glucose uptake (P<0.05), and also presented a higher insulin sensitivity, without changes in insulin binding. The two glucose carriers, GLUT1 and GLUT4, increased by 87% (P<0.05) and 90% (P<0. 05), respectively, in cells treated with T(3). Under non-insulin-stimulated conditions, plasma membrane fractions obtained from cells exposed to T(3) were enriched with both GLUT1 (3. 29+/-0.69 vs 1.20+/-0.29 arbitrary units (A.U.)/5 microg protein, P<0.05) and GLUT4 (3.50+/-1.16 vs 0.82+/-0.28 A.U./5 microg protein, P<0.03). The incubation of cells with insulin produced the translocation of both glucose transporters to plasma membranes, and again cells treated with T(3) presented a higher amount of GLUT1 and GLUT4 in the plasma membrane fractions (P<0.05 and P<0.03 respectively). These data indicate that T(3) has a direct stimulatory effect on glucose transport in 3T3-L1 adipocytes due to an increase in GLUT1 and GLUT4, and by favouring their partitioning to plasma membranes. The effect of T(3) on glucose uptake induced by insulin can also be explained by the high expression of both glucose transporters.

scholarly journals Thyroid receptor plasticity in striated muscle types: effects of altered thyroid state

1998 ◽  
Vol 274 (6) ◽  
pp. E1018-E1026 ◽  
Author(s):  
Fadia Haddad ◽  
Anqi X. Qin ◽  
Samuel A. McCue ◽  
Kenneth M. Baldwin
Keyword(s):  
Skeletal Muscle ◽  
Mrna Expression ◽  
Type I ◽  
Thyroid State ◽  
Muscle Types ◽  
Slow Twitch ◽  
Thyroid Receptor ◽  
Altered Thyroid ◽  
Fast Twitch ◽  
Skeletal Muscle Types

This study examined nuclear thyroid receptor (TR) maximum binding capacity (Bmax), dissociation constant ( K d), and TR isoform (α1, α2, β1) mRNA expression in rodent cardiac, “fast-twitch white,” “fast-twitch red,” and “slow-twitch red” muscle types as a function of thyroid state. These analyses were performed in the context of slow-twitch type I myosin heavy-chain (MHC) expression, a 3,5,3′-triiodothyronine (T3)-regulated gene that displays varying responsiveness to T3 in the above tissues. Nuclear T3 binding analyses show that the skeletal muscle types express more TRs per unit DNA than cardiac muscle, whereas the latter has a lower K d than the former. Altered thyroid state had little effect on either cardiac Bmax or K d, whereas hypothyroidism increased Bmax in the skeletal muscle types without affecting its K d. Cardiac muscle demonstrated the greatest mRNA signal of TR-β1 compared with the other muscle types, whereas the TR-α1mRNA signals were more abundant in the skeletal muscle types, especially fast-twitch red. Hyperthyroidism increased the ratio of β1 to α1 and decreased the ratio of α2- to α1+β1-mRNA signal across the muscle types, whereas hypothyroidism caused the opposite effects. The nuclear T3affinity correlated significantly with the TR-β1 mRNA expression but not with TR-α1 mRNA expression. Collectively, these findings suggest that, despite a divergent pattern of TR mRNA expression in the different muscle types, these patterns follow similar qualitative changes under altered thyroid state. Furthermore, TR expression pattern cannot account for the quantitative and qualitative changes in type I MHC expression that occur in the different muscle types.

scholarly journals Effects of spaceflight and thyroid deficiency on rat hindlimb development. II. Expression of MHC isoforms

2000 ◽  
Vol 88 (3) ◽  
pp. 904-916 ◽  
Author(s):  
G. R. Adams ◽  
F. Haddad ◽  
S. A. McCue ◽  
P. W. Bodell ◽  
M. Zeng ◽  
...  
Keyword(s):  
Skeletal Muscles ◽  
Weight Bearing ◽  
Type I ◽  
Thyroid State ◽  
Laboratory Rodents ◽  
Normal Expression ◽  
Slow Type ◽  
Slow Twitch ◽  
Fast Twitch ◽  
Fast Muscles

Both slow-twitch and fast-twitch muscles are undifferentiated after birth as to their contractile protein phenotype. Thus we examined the separate and combined effects of spaceflight (SF) and thyroid deficiency (TD) on myosin heavy chain (MHC) gene expression (protein and mRNA) in muscles of neonatal rats (7 and 14 days of age at launch) exposed to SF for 16 days. Spaceflight markedly reduced expression of the slow, type I MHC gene by ∼55%, whereas it augmented expression of the fast IIx and IIb MHCs in antigravity skeletal muscles. In fast muscles, SF caused subtle increases in the fast IIb MHC relative to the other adult MHCs. In contrast, TD prevented the normal expression of the fast MHC phenotype, particularly the IIb MHC, whereas TD maintained expression of the embryonic/neonatal MHC isoforms; this response occurred independently of gravity. Collectively, these results suggest that normal expression of the type I MHC gene requires signals associated with weight-bearing activity, whereas normal expression of the IIb MHC requires an intact thyroid state acting independently of the weight-bearing activities typically encountered during neonatal development of laboratory rodents. Finally, MHC expression in developing muscles is chiefly regulated by pretranslational processes based on the tight relationship between the MHC protein and mRNA data.

scholarly journals Strenuous Acute Exercise Induces Slow and Fast Twitch-Dependent NADPH Oxidase Expression in Rat Skeletal Muscle

Antioxidants ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 57 ◽  
Author(s):  
Juliana Osório Alves ◽  
Leonardo Matta Pereira ◽  
Igor Cabral Coutinho do Rêgo Monteiro ◽  
Luiz Henrique Pontes dos Santos ◽  
Alex Soares Marreiros Ferraz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Nadph Oxidase ◽  
Strenuous Exercise ◽  
Exercise Session ◽  
Type I ◽  
Fiber Types ◽  
Mrna Levels ◽  
Slow Twitch ◽  
Fast Twitch

The enzymatic complex Nicotinamide Adenine Dinucleotide Phosphate (NADPH) oxidase (NOx) may be the principal source of reactive oxygen species (ROS). The NOX2 and NOX4 isoforms are tissue-dependent and are differentially expressed in slow-twitch fibers (type I fibers) and fast-twitch fibers (type II fibers) of skeletal muscle, making them different markers of ROS metabolism induced by physical exercise. The aim of this study was to investigate NOx signaling, as a non-adaptive and non-cumulative response, in the predominant fiber types of rat skeletal muscles 24 h after one strenuous treadmill exercise session. The levels of mRNA, reduced glycogen, thiol content, NOx, superoxide dismutase, catalase, glutathione peroxidase activity, and PPARGC1α and SLC2A4 gene expression were measured in the white gastrocnemius (WG) portion, the red gastrocnemius (RG) portion, and the soleus muscle (SOL). NOx activity showed higher values in the SOL muscle compared to the RG and WG portions. The same was true of the NOX2 and NOX4 mRNA levels, antioxidant enzymatic activities, glycogen content. Twenty-four hours after the strenuous exercise session, NOx expression increased in slow-twitch oxidative fibers. The acute strenuous exercise condition showed an attenuation of oxidative stress and an upregulation of antioxidant activity through PPARGC1α gene activity, antioxidant defense adaptations, and differential gene expression according to the predominant fiber type. The most prominent location of detoxification (indicated by NOX4 activation) in the slow-twitch oxidative SOL muscle was the mitochondria, while the fast-twitch oxidative RG portion showed a more cytosolic location. Glycolytic metabolism in the WG portion suggested possible NOX2/NOX4 non-regulation, indicating other possible ROS regulation pathways.

Seasonal changes in plasma membrane glucose transporters enhance cryoprotectant distribution in the freeze-tolerant wood frog

1995 ◽  
Vol 73 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Patricia A. King ◽  
Mary N. Rosholt ◽  
Kenneth B. Storey
Keyword(s):  
Skeletal Muscle ◽  
Plasma Membrane ◽  
Glucose Transport ◽  
Seasonal Changes ◽  
Plasma Membranes ◽  
Glucose Transporter ◽  
Glucose Transporters ◽  
Wood Frog ◽  
Maximal Velocity ◽  
Liver Membranes

One of the critical adaptations for freeze tolerance by the wood frog, Rana sylvatica, is the production of large quantities of glucose as an organ cryoprotectant during freezing exposures. Glucose export from the liver, where it is synthesized, and its uptake by other organs is dependent upon carrier-mediated transport across plasma membranes by glucose-transporter proteins. Seasonal changes in the capacity to transport glucose across plasma membranes were assessed in liver and skeletal muscle of wood frogs; summer-collected (June) frogs were compared with autumn-collected (September) cold-acclimated (5 °C for 3–4 weeks) frogs. Plasma membrane vesicles prepared from liver of autumn-collected frogs showed 6-fold higher rates of carrier-mediated glucose transport than vesicles from summer-collected frogs, maximal velocity (Vmax) values for transport being 72 ± 14 and 12.0 + 2.9 nmol∙mg protein−1∙s−1, respectively (at 10 °C). However, substrate affinity constants for carrier-mediated glucose transport (K1/2) did not change seasonally. The difference in transport rates was due to greater numbers of glucose transporters in liver plasma membranes from autumn-collected frogs. The total number of transporter sites, as determined by cytochalasin B binding, was 8.5-fold higher in autumn than in summer. Glucose transporters in wood frog liver membranes cross-reacted with antibodies to the rat GluT-2 glucose transporter (the mammalian liver isoform), and Western blots further confirmed a large increase in transporter numbers in liver membranes from autumn- versus summer-collected frogs. By contrast with the liver, however, there were no seasonal changes in glucose-transporter activity or numbers in plasma membranes isolated from skeletal muscle. We conclude that an enhanced capacity for glucose transport across liver, but not muscle, plasma membranes during autumn cold-hardening is an important adaptation that anticipates the need for rapid export of cryoprotectant from liver during natural freezing episodes.

In vivo Ca2+ buffering capacity and microvascular oxygen pressures following muscle contractions in diabetic rat skeletal muscles: fiber-type specific effects

2015 ◽  
Vol 309 (2) ◽  
pp. R128-R137 ◽  
Author(s):  
Hiroaki Eshima ◽  
David C. Poole ◽  
Yutaka Kano
Keyword(s):  
Fiber Type ◽  
Recovery Period ◽  
Muscle Contractions ◽  
Diabetic Rat ◽  
Type I ◽  
Type Ii ◽  
Protein Levels ◽  
Slow Twitch ◽  
Fast Twitch

In Type 1 diabetes, skeletal muscle resting intracellular Ca2+ concentration ([Ca2+]i) homeostasis is impaired following muscle contractions. It is unclear to what degree this behavior is contingent upon fiber type and muscle oxygenation conditions. We tested the hypotheses that: 1) the rise in resting [Ca2+]i evident in diabetic rat slow-twitch (type I) muscle would be exacerbated in fast-twitch (type II) muscle following contraction; and 2) these elevated [Ca2+]i levels would relate to derangement of microvascular partial pressure of oxygen (PmvO2) rather than sarcoplasmic reticulum dysfunction per se. Adult male Wistar rats were divided randomly into diabetic (DIA: streptozotocin ip) and healthy (CONT) groups. Four weeks later extensor digitorum longus (EDL, predominately type II fibers) and soleus (SOL, predominately type I fibers) muscle contractions were elicited by continuous electrical stimulation (120 s, 100 Hz). Ca2+ imaging was achieved using fura 2-AM in vivo (i.e., circulation intact). DIA increased fatigability in EDL ( P < 0.05) but not SOL. In recovery, SOL [Ca2+]i either returned to its resting baseline within 150 s (CONT 1.00 ± 0.02 at 600 s) or was not elevated in recovery at all (DIA 1.03 ± 0.02 at 600 s, P > 0.05). In recovery, EDL CONT [Ca2+]i also decreased to values not different from baseline (1.06 ± 0.01, P > 0.05) at 600 s. In marked contrast, EDL DIA [Ca2+]i remained elevated for the entire recovery period (i.e., 1.23 ± 0.03 at 600 s, P < 0.05). The inability of [Ca2+]i to return to baseline in EDL DIA was not associated with any reduction of SR Ca2+-ATPase (SERCA) 1 or SERCA2 protein levels (both increased 30–40%, P < 0.05). However, PmvO2 recovery kinetics were markedly slowed in EDL such that mean PmvO2 was substantially depressed (CONT 27.9 ± 2.0 vs. DIA 18.4 ± 2.0 Torr, P < 0.05), and this behavior was associated with the elevated [Ca2+]i. In contrast, this was not the case for SOL ( P > 0.05) in that neither [Ca2+]i nor PmvO2 were deranged in recovery with DIA. In conclusion, recovery of [Ca2+]i homeostasis is impaired in diabetic rat fast-twitch but not slow-twitch muscle in concert with reduced PmvO2 pressures.

scholarly journals Myosin content of individual human muscle fibers isolated by laser capture microdissection

2016 ◽  
Vol 310 (5) ◽  
pp. C381-C389 ◽  
Author(s):  
Charles A. Stuart ◽  
William L. Stone ◽  
Mary E. A. Howell ◽  
Marianne F. Brannon ◽  
H. Kenton Hall ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Light Chain ◽  
Type I ◽  
Fiber Types ◽  
Myosin Heavy Chains ◽  
Heavy Chains ◽  
Slow Twitch ◽  
Type I Fibers ◽  
Fast Twitch ◽  
Laser Capture

Muscle fiber composition correlates with insulin resistance, and exercise training can increase slow-twitch (type I) fibers and, thereby, mitigate diabetes risk. Human skeletal muscle is made up of three distinct fiber types, but muscle contains many more isoforms of myosin heavy and light chains, which are coded by 15 and 11 different genes, respectively. Laser capture microdissection techniques allow assessment of mRNA and protein content in individual fibers. We found that specific human fiber types contain different mixtures of myosin heavy and light chains. Fast-twitch (type IIx) fibers consistently contained myosin heavy chains 1, 2, and 4 and myosin light chain 1. Type I fibers always contained myosin heavy chains 6 and 7 (MYH6 and MYH7) and myosin light chain 3 (MYL3), whereas MYH6, MYH7, and MYL3 were nearly absent from type IIx fibers. In contrast to cardiomyocytes, where MYH6 (also known as α-myosin heavy chain) is seen solely in fast-twitch cells, only slow-twitch fibers of skeletal muscle contained MYH6. Classical fast myosin heavy chains (MHC1, MHC2, and MHC4) were present in variable proportions in all fiber types, but significant MYH6 and MYH7 expression indicated slow-twitch phenotype, and the absence of these two isoforms determined a fast-twitch phenotype. The mixed myosin heavy and light chain content of type IIa fibers was consistent with its role as a transition between fast and slow phenotypes. These new observations suggest that the presence or absence of MYH6 and MYH7 proteins dictates the slow- or fast-twitch phenotype in skeletal muscle.

Adaptive responses of hypertrophying skeletal muscle to endurance training

1996 ◽  
Vol 81 (2) ◽  
pp. 665-672 ◽  
Author(s):  
J. Stone ◽  
T. Brannon ◽  
F. Haddad ◽  
A. Qin ◽  
K. M. Baldwin
Keyword(s):  
Endurance Training ◽  
Citrate Synthase ◽  
Compensatory Hypertrophy ◽  
Type I ◽  
Adaptive Responses ◽  
Endurance Running ◽  
Proportional Increase ◽  
Muscle Types ◽  
Slow Twitch ◽  
Fast Twitch

We determined the effects of 6 wk of endurance running on citrate synthase (CS) activity and myosin heavy chain (MHC) expression in bilateral surgical-overloaded rodent fast-twitch plantaris and slow-twitch soleus muscles. The overload induced significant hypertrophy in both muscle types, and this response was enhanced by endurance training. The overload-induced compensatory hypertrophy was accompanied by a proportional increase in muscle CS content. Although endurance training produced significant increases in CS concentration in either muscle type of the normal-trained groups (P < 0.05), it was not effective in causing similar changes in the overloaded trained muscles. Also, overload of either the sedentary or trained groups produced an increase in slower MHC isoforms (i.e., type I in the soleus and types I and IIa in the plantaris) and a concomitant decrease in the faster MHC isoforms (type IIa in the soleus and IIb in the plantaris; P < 0.05), whereas endurance training alone produced the opposite effect, especially in the plantaris. Collectively, these data suggest that 1) increments in muscle oxidative enzyme content due to endurance training are compromised when a hypertrophying process is occurring concomitantly; and 2) the relative loading state imposed on the muscle during repetitive locomotor activity is critical in regulating the pattern of MHC plasticity.

Low sulphur content in atrophied human skeletal muscle: an electron probe X-ray microanalytical study

1978 ◽  
Vol 36 (2) ◽  
pp. 634-635
Author(s):  
R. Wróblewski ◽  
W. Gremski ◽  
G. M. Roomans ◽  
R. Nordemar ◽  
L. Edström
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Control Group ◽  
Muscle Fibres ◽  
Type I ◽  
Tissue Preparation ◽  
Type Ii ◽  
Slow Twitch ◽  
Fibre Typing ◽  
Fast Twitch

Many diseases of the human skeletal muscle involve an atrophy of the muscle fibres. In some cases mainly one of the fibre types is affected. The fibre typing system used in this study is that of Padykula and Herman, 1955 and distinguishes between type I fibres which presumably correspond to the slow-twitch fibres and type II fibres which are the fast-twitch fibres. The type II fibres can be divided into type II A, II B and II C fibres. Recent advances in instrumentation and tissue preparation have permitted an investigation of the elemental composition of individual muscle fibres of known fibre type with the aim of comparing healthy and atrophied muscle fibres.In this study we have examined ten patients suffering from rheumatoid arthritis, two patients suffering from Parkinson's disease and two patients with upper motor lesions. As a control group we have examined muscles from eight healthy controls of the same age.

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