Absence of Insulin Receptor Substrate-1 Expression Does Not Alter GLUT1 or GLUT4 Abundance or Contraction-Stimulated Glucose Uptake by Mouse Skeletal Muscle

The purpose of this study was to determine the influence of insulin receptor substrate-1 (IRS-1) expression on GLUT1 and GLUT4 glucose transporter protein abundance, contraction-stimulated glucose uptake, and contraction-induced glycogen depletion by skeletal muscle. Mice (6 months old) from three genotypes were studied: wild-type (IRS-1^+/+), heterozygous (IRS-1^+/-) for the null allele, and IRS-1 knockouts (IRS-1^-/-) lacking a functional IRS-1 gene. In situ muscle contraction was induced (electrical stimulation of sciatic nerve) in one hindlimb using contralateral muscles as controls. Soleus and extensor digitorum longus were dissected and 2-deoxyglucose uptake was measured in vitro. 2-Deoxyglucose uptake was higher in basal muscles (no contractions) from IRS-1^-/- vs. both other genotypes. Contraction-stimulated 2-deoxyglucose uptake and glycogen depletion did not differ among genotypes. Muscle IRS-1 protein was undetectable for IRS-1^-/- mice, and values were approximately 40 % lower in IRS-1^+/- than in IRS-1^+/+ mice. No difference was found in IRS-1^+/+ compared to IRS-1^-/- groups regarding muscle abundance of GLUT1 and GLUT4. Substantial reduction or elimination of IRS-1 did not alter the hallmark effects of contractions on muscle carbohydrate metabolism - activation of glucose uptake and glycogen depletion.

Exercise - Glucose Transport - Insulin Signaling - Contractile Activity - Glycogen

References

• 1 Goodyear L J, Kahn B B. Exercise, glucose transport, and insulin sensitivity.  Annu Rev Med. 1998;  49 235-261
• 2 Holloszy J O, Hansen P A. Regulation of glucose transport into skeletal muscle.  Rev Physiol, Biochem Pharmacol. 1996;  128 99-193
• 3 Kahn C R. Insulin action, diabetogenes, and the cause of type II diabetes.  Diabetes. 1994;  43 1066-1084
• 4 Patti M-E, Sun X-J, Bruening J C, Araki E, Lipes M A, White M F, Kahn C R. 4PS/Insulin receptor substrate (IRS)-2 is the alternative substrate of the insulin receptor in IRS-1-deficient mice.  J Biol Chem. 1995;  270 24 670-24 673
• 5 Yeh J-I, Gulve E A, Rameh L, Birnbaum M J. The effects of wortmannin on rat skeletal muscle.  J Biol Chem. 1995;  270 2107-2111
• 6 Gazdag A C, Dumke C L, Kahn C R, Cartee G D. Calorie restriction increases insulin-stimulated glucose transport in skeletal muscle from IRS-1 knockout mice.  Diabetes. 1999;  48 1930-1936
• 7 Lund S, Flyvbjerg A, Holman G D, Larsen F S, Pedersen O, Schmitz O. Comparative effects of IGF-I and insulin on the glucose transporter system in rat muscle.  Am J Physiol. 1994;  267 E461-E466
• 8 Satoh S, Nishimura H, Clark A E, Kozka I J, Vannucci S J, Simpson I A, Quon M J, Cushman S W, Holman G D. Use of bismannose photolabel to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action.  J Biol Chem. 1993;  268 17 820-17 829
• 9 Derave W, Hansen B F, Lund S, Kristiansen S, Richter E A. Muscle glycogen content affects insulin-stimulated glucose transport and protein kinase B activity.  Am J Physiol. 2000;  279 E947-E955
• 10 Araki E, Lipes M A, Patti M-E, Bruning J C, Haag B  III, Johnson R S, Kahn C R. Alternative pathway of insulin signalling in mice with targeted disruption of the IRS-1 gene.  Nature. 1994;  372 186-190
• 11 Gulve E A, Ren J M, Marshall B A, Gao J, Hansen P A, Holloszy J O, Mueckler M. Glucose transport activity in skeletal muscles from transgenic mice overexpressing GLUT1.  J Biol Chem. 1994;  269 18 366-18 370
• 12 Hansen P A, Gulve E A, Holloszy J O. Suitability of 2-deoxyglucose for in vitro measurement of glucose transport activity in skeletal muscle.  J Appl Physiol. 1994;  76 979-985
• 13 Passonneau J V, Lauderdale V R. A comparison of three methods of glycogen measurement in tissues.  Anal Biochem. 1974;  60 405-412
• 14 Dean D J, Brozinick J T , Cushman S W, Cartee G D. Calorie restriction increases cell surface GLUT4 in insulin-stimulated skeletal muscle.  Am J Physiol. 1998;  275 E957-E964
• 15 Yamauchi T, Tobe K, Tamemoto H, Ueki K, Kaburagi Y, Yamamoto-Honda R, Takahashi Y, Yoshizawa F, Aizawa S, Akanuma Y, Soneberg N, Yazaki Y, Kadowaki T. Insulin signalling and insulin actions in the muscles and livers of insulin-resistant, insulin receptor substrate 1-deficient mice.  Mol Cell Biol. 1996;  16 3074-3084
• 16 Goodman M, Dluz S, McElaney M, Belur E, Ruderman N. Glucose uptake and insulin sensitivity in rat muscle: changes during 3 - 96 weeks of age.  Am J Physiol. 1983;  244 E93-E100
• 17 Kulkarni R N, Almind K, Quinn R L, Eisenman D, Kahn C R. Dramatic differences in phenotype in mice double heterozygous for insulin receptor (ER) and insulin receptor substrate-1 (IRS-1) on different genetic backgrounds.  Diabetes. 2001;  50 (Suppl. 2) A317 (Abstract)
• 18 Bohni R, Riesgo-Escovar J, Oldham S, Brogiolo W, Stocker H, Andruss B F, Beckingham K, Hafen E. Autonomous control of cell and organ size by CHICO, a Drosophila homolog of vertebrate IRS-1 - 4.  Cell. 1999;  97 865-875
• 19 Previs S F, Withers D J, Ren J-M, White M F, Shulman G I. Contrasting effects of IRS-1 vs. IRS-2 gene disruption on carbohydrate and lipid metabolism in vitro. .  J Biol Chem. 2000;  275 38 990-38 994
• 20 Higaki Y, Wojtaszewski J FP, Hirshman M F, Withers D J, Towery H, White M F, Goodyear L J. Insulin receptor substrate-2 is not necessary for insulin- and exercise-stimulated glucose transport in skeletal muscle.  J Biol Chem. 1999;  274 20 791-20 795