Summary
Glycogen phosphorylase regulates the breakdown of glycogen into glucose, but as previous studies have demonstrated, the control of glycogen metabolism becomes deregulated in diabetes mellitus. Messenger RNA levels encoding several different proteins are altered in skeletal muscle biopsies of patients with insulin-dependent and non-insulin-dependent diabetes. The possible alteration of expression of the gene encoding the skeletal muscle isoform of glycogen phosphorylase during diabetes has not previously been investigated. We examined the effect of streptozotocin-induced diabetes and insulin treatment on glycogen phosphorylase mRNA in rat skeletal muscle; glycogen phosphorylase mRNA levels were elevated in diabetic rat muscle tissue, but were partially suppressed in diabetic rat muscle following insulin treatment. To distinguish between the effects of insulin and counter-regulatory hormones on glycogen phosphorylase mRNA levels, we employed differentiating rat L6 myoblasts in culture. Insulin stimulated the accumulation of glycogen phosphorylase mRNA as determined by Northern blot analysis. Moreover, insulin and dibutyryl cAMP stimulated expression of a transiently transfected chloramphenicol acetyl transferase reporter gene under the control of the muscle glycogen phosphorylase promoter in differentiating myotubes in culture, suggesting that the effects of insulin and counter-regulatory hormones on glycogen phosphorylase mRNA are at the level of transcription. These results suggest that insulin and epinephrine may participate in the induction of the glycogen phosphorylase gene during myogenesis; moreover, activation of this gene in muscle tissue may be a contributing factor in impaired glycogen storage during uncontrolled diabetes.
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Abbreviations
- CAT:
-
Chloramphenicol acetyl transferase
- GP:
-
glycogen phosphorylase
- GS:
-
Glycogen synthase
- STZ:
-
streptozotocin
- cAMP:
-
cyclic AMP
- db cAMP:
-
dibutyryl cAMP
- hGH:
-
human growth hormone
- EF1γ:
-
elongation factor gamma
- bp:
-
base pair
- MEM:
-
modified essential medium
- DMEM:
-
Dulbecco's modified essential medium
- CREB:
-
cAMP Response element binding protein
References
Kelley D, Mitrakou A, Marsh H et al (1988) Skeletal muscle glycolysis, oxidation, and storage of an oral glucose load. J Clin Invest 81: 1563–1571
DeFronzo R, Gunnarsson R, Bjorkman O, Olsson M, Wahren J (1985) Effects of insulin on peripheral and splanchnic glucose metabolism in noninsulin-dependent (type II) diabetes mellitus. J Clin Invest 76: 149–155
Sutherland EW, Oye I, Butcher RW (1965) The action of epinephrine and the role of the adenyl cyclase system in hormone action. Recent Progr Hormone Res 21: 623–646
Craig JW, Larner J (1964) Influence of epinephrine and insulin on uridine diphosphate glucose-alpha-glucan transferase and phosphorylase in muscle. Nature 202: 971–973
Johnson AB, Argyraki M, Thow JC, Broughton D, Jones IR, Taylor R (1990) Effects of intensive dietary treatment on insulin-stimulated skeletal muscle glycogen synthase activation and insulin secretion in newly presenting type 2 diabetic patients. Diabet Med 7: 420–428
Bogardus C, Lillioja S, Stone K, Mott D (1984) Correlation between muscle glycogen synthase activity and in vivo insulin action in man. J Clin Invest 73: 1185–1190
Thorburn AW, Gumbiner B, Bulacan F, Brechtel G, Henry RR (1991) Multiple defects in muscle glycogen synthase activity contribute to reduced glycogen synthesis in non-insulin dependent diabetes mellitus. J Clin Invest 87: 489–495
Bjorbaek C, Echwald SM, Hubricht P et al (1994) Genetic variants in promoters and coding regions of the muscle glycogen synthase and the insulin-responsive GLUT 4 genes in NIDDM. Diabetes 43: 976–983
Freymond D, Bogardus C, Okubo M, Stone K, Mott D (1988) Impaired insulin-stimulated muscle glycogen synthase activation in vivo in man is related to low fasting glycogen synthase phosphatase activity. J Clin Invest 82: 1503–1509
Kida Y, Esposito-Del Puente A, Bogardus C, Mott DM (1990) Insulin resistance is associated with reduced fasting and insulin-stimulated glycogen synthase phosphatase activity in human skeletal muscle. J Clin Invest 85: 476–481
Walkenbach RJ, Hazen R, Larner J (1980) Hormonal regulation of glycogen synthase. Insulin decreases protein kinase sensitivity to cyclic AMP. Biochim Biophys Acta 629: 421–430
Okubo M, Bogardus C, Lillioja S, Mott DM (1989) Adenosine 3′, 5′-monophosphate-dependent protein kinase activity decreases in human muscle after insulin infusion. J Clin Endocrinol Metab 69: 798–803
Torres HN, Marechal LR, Bernard E, Belocopitow E (1968) Control of muscle glycogen phosphorylase activity by insulin. Biochim Biophys Acta 156: 206–209
Shen LC, Villar-Palisi C, Larner J (1970) Hormonal alteration of protein kinase sensitivity to 3′ 5′-cyclic AMP. Physiol Chem Phys 2: 536–544
Kida Y, Nyomba BL, Bogardus C, Mott DM (1991) Defective insulin response of cyclic adenosine monophosphate-dependent protein kinase in insulin-resistant humans. J Clin Invest 87: 673–679
Yaffe D (1968) Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci USA 61: 477–483
Beguinot F, Kahn CR, Moses AC, Smith RJ (1986) The development of insulin receptors and responsiveness is an early marker of differentiation in the muscle cell line L6. Endocrinology 118: 446–455
Loeken MR, Khoury G, Brady J (1986) Stimulation of the adenovirus E2 promoter by simian virus 40 T antigen or E1A occurs by different mechanisms. Mol Cell Biol 6: 2020–2026
Loeken MR (1993) Effects of mutation of the CREB binding site of the somatostatin promoter on cyclic AMP responsiveness in CV-1 cells. Gene Expression 3: 253–264
Selden RF (1987) Assay for human growth hormone. Current protocols in molecular biology. Greene Publishing Associates, New York, 9.7. 1–2
Gorman CM, Moffat LF, Howard BH (1983) Recombinant genomes which express chloramphenicol acetyl transferase in mammalian cells. Mol Cell Biol 2: 1044–1051
Saad MJA, Araki E, Miralpeix M, Rothenberg PL, White MF, Kahn CR (1992) Regulation of insulin receptor substrate-1 in liver and muscle of animal models of insulin resistance. J Clin Invest 90: 1839–1849
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by guanidinium thiocyanate-pnenol-chloroform extraction. Anal Biochem 162: 156–159
Reynet C, Kahn CR (1993) Rad: a member of the ras family overexpressed in muscle of type II diabetic humans. Science 262: 1441–1444
Hwang PK, See YP, Vincentini AM, Powers MA, Fletterick RJ, Crerar MM (1985) Comparative sequence analysis of rat, rabbit, and human muscle glycogen phosphorylase cDNAs. Eur J Biochem 152: 267–274
Lockyer JM, McCracken Jr JB (1991) Identification of a tissue-specific regulatory element within the human muscle glycogen phosphorylase gene. J Biol Chem 266: 20262–20269
Selden RF, Burke-Howie K, Rowe ME, Goodman HM, Moore DD (1986) Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol Cell Biol 6: 3173–3179
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Kahn CR, Reynet C (1994) Identification of diabetes-related genes by subtraction cloning. Molecular biology of diabetes. Humana Press, Totowa, New Jersey, pp 51–77
David E, Crerar MM (1986) Quantitation of muscle glycogen phosphorylase mRNA and enzyme amounts in adult rat tissues. Biochim Biophys Acta 880: 78–90
Ewton DA, Falen SL, Florini JR (1987) The type II insulinlike growth factor (IGF) receptor has low affinity for IGF-I analogs: pleiotypic actions of IGFs on myoblasts are apparently mediated by the type I receptor. Endocrinology 120: 115–123
Iujvidin S, Fuchs O, Nudel U, Yaffe D (1990) SV40 immortalizes myogenic cells: DNA synthesis and mitosis in differentiating myotubes. Differentiation 43: 192–203
O'Brien RM, Bonovich MT, Forest CD, Granner DK (1991) Signal transduction convergence: phorbol esters and insulin inhibit phosphoenolpyruvate carboxykinase gene transcription through the same 10-base-pair sequence. Proc Natl Acad Sci USA 88: 6580–6584
Magnuson MA, Andreone TL, Printz RL, Kach S, Granner DK (1989) The glucokinase gene: structure and regulation by insulin. Proc Natl Acad Sci USA 86: 4838–4842
Hoeffler JP, Meyer TE, Yun Y, Jameson JL, Habener JF (1988) Cyclic AMP-responsive DNA-binding protein: structure based on a cloned placental cDNA. Science 242: 1430–1433
Gonzalez GA, Yamamoto KK, Fischer WH et al (1989) A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence. Nature 337: 749–752
Quinn PG (1994) Inhibition by insulin of protein kinase A-induced transcription of the phosphoenolpyruvate carboxykinase gene-mediation by the activation domain of cAMP response element-binding protein (CREB) and factors bound to the TATA box. J Biol Chem 269: 14375–14378
Montminy MR, Sevarino KA, Wagner JA, Mandel G, Goodman RH (1986) Identification of a cyclic-AMP-responsive element within the rat somatostatin gene. Proc Natl Acad Sci USA 83: 6682–6686
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Reynet, C., Kahn, C.R. & Loeken, M.R. Expression of the gene encoding glycogen phosphorylase is elevated in diabetic rat skeletal muscle and is regulated by insulin and cyclic AMP. Diabetologia 39, 183–189 (1996). https://doi.org/10.1007/BF00403961
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DOI: https://doi.org/10.1007/BF00403961