Muscle-specific PPARgamma-deficient mice develop increased adiposity and insulin resistance but respond to thiazolidinediones.

Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) by thiazolidinediones (TZDs) improves insulin resistance by increasing insulin-stimulated glucose disposal in skeletal muscle. It remains debatable whether the effect of TZDs on muscle is direct or indirect via adipose tissue. We therefore generated mice with muscle-specific PPARgamma knockout (MuPPARgammaKO) using Cre/loxP recombination. Interestingly, MuPPARgammaKO mice developed excess adiposity despite reduced dietary intake. Although insulin-stimulated glucose uptake in muscle was not impaired, MuPPARgammaKO mice had whole-body insulin resistance with a 36% reduction (P < 0.05) in the glucose infusion rate required to maintain euglycemia during hyperinsulinemic clamp, primarily due to dramatic impairment in hepatic insulin action. When placed on a high-fat diet, MuPPARgammaKO mice developed hyperinsulinemia and impaired glucose homeostasis identical to controls. Simultaneous treatment with TZD ameliorated these high fat-induced defects in MuPPARgammaKO mice to a degree identical to controls. There was also altered expression of several lipid metabolism genes in the muscle of MuPPARgammaKO mice. Thus, muscle PPARgamma is not required for the antidiabetic effects of TZDs, but has a hitherto unsuspected role for maintenance of normal adiposity, whole-body insulin sensitivity, and hepatic insulin action. The tissue crosstalk mediating these effects is perhaps due to altered lipid metabolism in muscle.

[1]  Johan Auwerx,et al.  A Pro12Ala substitution in PPARγ2 associated with decreased receptor activity, lower body mass index and improved insulin sensitivity , 1998, Nature Genetics.

[2]  E S Lander,et al.  The common PPARgamma Pro12Ala polymorphism is associated with decreased risk of type 2 diabetes. , 2000, Nature genetics.

[3]  D. James,et al.  A potent in vivo effect of ciglitazone on muscle insulin resistance induced by high fat feeding of rats. , 1989, Metabolism: clinical and experimental.

[4]  J. Morgan Myogenicity in vitro and in vivo of mouse muscle cells separated on discontinuous Percoll gradients , 1988, Journal of the Neurological Sciences.

[5]  B. Wolffenbuttel,et al.  Plasma FFA utilization and fatty acid-binding protein content are diminished in type 2 diabetic muscle. , 2000, American journal of physiology. Endocrinology and metabolism.

[6]  J. Auwerx,et al.  Tissue Distribution and Quantification of the Expression of mRNAs of Peroxisome Proliferator–Activated Receptors and Liver X Receptor-α in Humans: No Alteration in Adipose Tissue of Obese and NIDDM Patients , 1997, Diabetes.

[7]  D. Kelly,et al.  A critical role for the peroxisome proliferator-activated receptor alpha (PPARalpha) in the cellular fasting response: the PPARalpha-null mouse as a model of fatty acid oxidation disorders. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[8]  W. Saris,et al.  Impaired oxidation of plasma-derived fatty acids in type 2 diabetic subjects during moderate-intensity exercise. , 2000, Diabetes.

[9]  J. Flatt,et al.  Effect of dietary fat content on the incidence of obesity among ad libitum fed mice. , 1985, International journal of obesity.

[10]  B. Spiegelman,et al.  PPARgamma : a nuclear regulator of metabolism, differentiation, and cell growth. , 2001, The Journal of biological chemistry.

[11]  S. Mudaliar,et al.  Peroxisome Proliferator-Activated Receptor (PPAR) γ and Retinoid X Receptor (RXR) agonists have complementary effects on glucose and lipid metabolism in human skeletal muscle , 2001, Diabetologia.

[12]  C. Glass,et al.  Conditional Disruption of the Peroxisome Proliferator-Activated Receptor γ Gene in Mice Results in Lowered Expression of ABCA1, ABCG1, and apoE in Macrophages and Reduced Cholesterol Efflux , 2002, Molecular and Cellular Biology.

[13]  G. Cooney,et al.  Peroxisome Proliferator—Activated Receptor (PPAR)-α Activation Lowers Muscle Lipids and Improves Insulin Sensitivity in High Fat—Fed Rats Comparison With PPAR-γ Activation , 2001 .

[14]  K. Umesono,et al.  Troglitazone increases the number of small adipocytes without the change of white adipose tissue mass in obese Zucker rats. , 1998, The Journal of clinical investigation.

[15]  K. Heidenreich,et al.  In vitro studies on the action of CS-045, a new antidiabetic agent. , 1990, Metabolism: clinical and experimental.

[16]  B. Spiegelman,et al.  C/EBPalpha induces adipogenesis through PPARgamma: a unified pathway. , 2002, Genes & development.

[17]  J. Holder,et al.  Insulin and rosiglitazone regulation of lipolysis and lipogenesis in human adipose tissue in vitro. , 2002, Diabetes.

[18]  R. Evans,et al.  Improved insulin-sensitivity in mice heterozygous for PPAR-gamma deficiency. , 2000, The Journal of clinical investigation.

[19]  J. Kolls,et al.  Tumor necrosis factor-alpha induces hepatic insulin resistance in obese Zucker (fa/fa) rats via interaction of leukocyte antigen-related tyrosine phosphatase with focal adhesion kinase. , 2000, Diabetes.

[20]  K. Chien,et al.  PPARγ Is Required for Placental, Cardiac, and Adipose Tissue Development , 1999 .

[21]  E. Ravussin,et al.  Insulin resistance and insulin secretory dysfunction as precursors of non-insulin-dependent diabetes mellitus. Prospective studies of Pima Indians. , 1993, The New England journal of medicine.

[22]  G. Cooney,et al.  Peroxisome proliferator-activated receptor (PPAR)-alpha activation lowers muscle lipids and improves insulin sensitivity in high fat-fed rats: comparison with PPAR-gamma activation. , 2001, Diabetes.

[23]  R. N. Bergman,et al.  Role of glucose and insulin resistance in development of type 2 diabetes mellitus: results of a 25-year follow-up study , 1992, The Lancet.

[24]  W. Wilfinger,et al.  Dissociation and enrichment of rat atrial myocytes containing atrial natriuretic factor (ANF). , 1986, Hormone research.

[25]  S. Mudaliar,et al.  Peroxisome proliferator-activated receptor (PPAR) gamma and retinoid X receptor (RXR) agonists have complementary effects on glucose and lipid metabolism in human skeletal muscle. , 2001, Diabetologia.

[26]  R. Evans,et al.  Improved insulin-sensitivity in mice heterozygous for PPAR-γ deficiency , 2000 .

[27]  R. Larkins,et al.  Evolution of Insulin Resistance in New Zealand Obese Mice , 1991, Diabetes.

[28]  K. Moulder,et al.  Sulfated Glycans Stimulate Endocytosis of the Cellular Isoform of the Prion Protein, PrPC, in Cultured Cells (*) , 1995, The Journal of Biological Chemistry.

[29]  J. Berger,et al.  Role of skeletal muscle in thiazolidinedione insulin sensitizer (PPARgamma agonist) action. , 1998, Endocrinology.

[30]  M. Lazar,et al.  The hormone resistin links obesity to diabetes , 2001, Nature.

[31]  K. Dudzinski,et al.  A technique for the combination of clearing, staining, and injecting small mammals. , 1990, Stain technology.

[32]  B. Brewer,et al.  Liver-specific disruption of PPARgamma in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes. , 2003, The Journal of clinical investigation.

[33]  B. Spiegelman PPAR-gamma: adipogenic regulator and thiazolidinedione receptor. , 1998, Diabetes.

[34]  T. Willson,et al.  Comprehensive Messenger Ribonucleic Acid Profiling Reveals That Peroxisome Proliferator-Activated Receptor γ Activation Has Coordinate Effects on Gene Expression in Multiple Insulin-Sensitive Tissues. , 2001, Endocrinology.

[35]  Margaret S. Wu,et al.  Role of Skeletal Muscle in Thiazolidinedione Insulin Sensitizer ( PPAR g Agonist ) Action , 1998 .

[36]  M. Vranic,et al.  A moderate decline in specific activity does not lead to an underestimation of hepatic glucose production during a glucose clamp. , 1996, Metabolism: clinical and experimental.

[37]  L. Mandarino,et al.  Fuel selection in human skeletal muscle in insulin resistance: a reexamination. , 2000, Diabetes.

[38]  S. O’Rahilly,et al.  Dominant negative mutations in human PPARγ associated with severe insulin resistance, diabetes mellitus and hypertension , 1999, Nature.

[39]  J. Auwerx,et al.  Induction of the Fatty Acid Transport Protein 1 and Acyl-CoA Synthase Genes by Dimer-selective Rexinoids Suggests That the Peroxisome Proliferator-activated Receptor-Retinoid X Receptor Heterodimer Is Their Molecular Target* , 2000, The Journal of Biological Chemistry.

[40]  Y. Terauchi,et al.  The mechanisms by which both heterozygous peroxisome proliferator-activated receptor gamma (PPARgamma) deficiency and PPARgamma agonist improve insulin resistance. , 2001, The Journal of biological chemistry.

[41]  C. Kahn,et al.  Redistribution of substrates to adipose tissue promotes obesity in mice with selective insulin resistance in muscle. , 2000, The Journal of clinical investigation.

[42]  Jens C. Brüning,et al.  Dilated cardiomyopathy and atrioventricular conduction blocks induced by heart-specific inactivation of mitochondrial DNA gene expression , 1999, Nature Genetics.

[43]  M. Jimenez-Linan,et al.  Peroxisome proliferator-activated receptor gene expression in human tissues. Effects of obesity, weight loss, and regulation by insulin and glucocorticoids. , 1997, The Journal of clinical investigation.

[44]  R. Evans,et al.  PPAR gamma is required for placental, cardiac, and adipose tissue development. , 1999, Molecular cell.

[45]  K. Petersen,et al.  Mechanism of troglitazone action in type 2 diabetes. , 2000, Diabetes.

[46]  C. Kahn,et al.  Glucose toxicity and the development of diabetes in mice with muscle-specific inactivation of GLUT4. , 2001, The Journal of clinical investigation.

[47]  J. Auwerx,et al.  Pioglitazone Induces In Vivo Adipocyte Differentiation in the Obese Zucker fa/fa Rat , 1997, Diabetes.

[48]  J. Olefsky,et al.  Skeletal muscle peroxisome proliferator- activated receptor-gamma expression in obesity and non- insulin-dependent diabetes mellitus. , 1998, The Journal of clinical investigation.

[49]  L. Hennighausen,et al.  Loss of the Peroxisome Proliferation-activated Receptor gamma (PPARγ) Does Not Affect Mammary Development and Propensity for Tumor Formation but Leads to Reduced Fertility* , 2002, The Journal of Biological Chemistry.

[50]  Tatsuya Hayashi,et al.  Evidence for 5′AMP-Activated Protein Kinase Mediation of the Effect of Muscle Contraction on Glucose Transport , 1998, Diabetes.

[51]  J. Peters,et al.  Altered Constitutive Expression of Fatty Acid-metabolizing Enzymes in Mice Lacking the Peroxisome Proliferator-activated Receptor α (PPARα)* , 1998, The Journal of Biological Chemistry.

[52]  M. Vranic,et al.  Determinants of glucose turnover in the pathophysiology of diabetes: an in vivo analysis in diabetic dogs. , 1996, Diabetes & metabolism.

[53]  C. Kahn,et al.  Targeted disruption of the glucose transporter 4 selectively in muscle causes insulin resistance and glucose intolerance , 2000, Nature Medicine.

[54]  J. Filmus,et al.  Transforming Growth Factor-α Prevents Detachment-induced Inhibition of c-Src Kinase Activity, Bcl-XLDown-regulation, and Apoptosis of Intestinal Epithelial Cells* , 2001, The Journal of Biological Chemistry.

[55]  J. Lehmann,et al.  An Antidiabetic Thiazolidinedione Is a High Affinity Ligand for Peroxisome Proliferator-activated Receptor γ (PPARγ) (*) , 1995, The Journal of Biological Chemistry.

[56]  W. Kraus,et al.  Fatty Acid Homeostasis and Induction of Lipid Regulatory Genes in Skeletal Muscles of Peroxisome Proliferator-activated Receptor (PPAR) α Knock-out Mice , 2002, The Journal of Biological Chemistry.

[57]  M. Lazar,et al.  Resistin and obesity-associated insulin resistance , 2002, Trends in Endocrinology & Metabolism.

[58]  J. Peters,et al.  Altered constitutive expression of fatty acid-metabolizing enzymes in mice lacking the peroxisome proliferator-activated receptor alpha (PPARalpha). , 1998, The Journal of biological chemistry.

[59]  A. Pfeiffer,et al.  Obesity associated with a mutation in a genetic regulator of adipocyte differentiation. , 1998, The New England journal of medicine.

[60]  J. Lehmann,et al.  An antidiabetic thiazolidinedione is a high affinity ligand for peroxisome proliferator-activated receptor gamma (PPAR gamma). , 1995, The Journal of biological chemistry.

[61]  B. Spiegelman,et al.  PPARγ: a Nuclear Regulator of Metabolism, Differentiation, and Cell Growth* , 2001, The Journal of Biological Chemistry.

[62]  M. Matsuda,et al.  PPARgamma ligands increase expression and plasma concentrations of adiponectin, an adipose-derived protein. , 2001, Diabetes.

[63]  L. Chao,et al.  Adipose tissue is required for the antidiabetic, but not for the hypolipidemic, effect of thiazolidinediones. , 2000, The Journal of clinical investigation.

[64]  T. Ishihara,et al.  Actions of novel antidiabetic thiazolidinedione, T‐174, in animal models of non‐insulin‐dependent diabetes mellitus (NIDDM) and in cultured muscle cells , 1998, British journal of pharmacology.

[65]  R. DeFronzo,et al.  The Effect of Insulin on the Disposal of Intravenous Glucose: Results from Indirect Calorimetry and Hepatic and Femoral Venous Catheterization , 1981, Diabetes.

[66]  D. Archer,et al.  Monoamine oxidase activities in dissociated cell fractions from rat skeletal muscle , 1986, The Journal of pharmacy and pharmacology.

[67]  P. Scherer,et al.  The adipocyte-secreted protein Acrp30 enhances hepatic insulin action , 2001, Nature Medicine.

[68]  C. Kahn,et al.  A muscle-specific insulin receptor knockout exhibits features of the metabolic syndrome of NIDDM without altering glucose tolerance. , 1998, Molecular cell.

[69]  P. Chambon,et al.  Conditional site-specific recombination in mammalian cells using a ligand-dependent chimeric Cre recombinase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[70]  G. Cooney,et al.  Expression of genes involved in lipid metabolism correlate with peroxisome proliferator-activated receptor gamma expression in human skeletal muscle. , 2000, The Journal of clinical endocrinology and metabolism.

[71]  Y. Terauchi,et al.  The Mechanisms by Which Both Heterozygous Peroxisome Proliferator-activated Receptor γ (PPARγ) Deficiency and PPARγ Agonist Improve Insulin Resistance* , 2001, The Journal of Biological Chemistry.

[72]  J. McGarry,et al.  Prolonged inhibition of muscle carnitine palmitoyltransferase-1 promotes intramyocellular lipid accumulation and insulin resistance in rats. , 2001, Diabetes.

[73]  C. Burant,et al.  Troglitazone action is independent of adipose tissue. , 1997, The Journal of clinical investigation.

[74]  G R Stark,et al.  Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[75]  M. Loeken,et al.  Reduced expression of Pax-3 is associated with overexpression of cdc46 in the mouse embryo , 1998, Development Genes and Evolution.

[76]  A. Hevener,et al.  Thiazolidinedione treatment prevents free fatty acid-induced insulin resistance in male wistar rats. , 2001, Diabetes.