Journal of Comparative Physiology B

, Volume 165, Issue 1, pp 62–70 | Cite as

Role of glucose and insulin in regulating glycogen synthase and phosphorylase activities in rainbow trout hepatocytes

  • C. Pereira
  • M. M. Vijayan
  • K. B. Storey
  • R. A. Jones
  • T. W. Moon
Original Paper

Abstract

This study, using 13C nuclear magnetic resonance spectroscopy showed enrichment of glycogen carbon (C1) from 13C-labelled (C1) glucose indicating a direct pathway for glycogen synthesis from glucose in rainbow trout (Oncorhynchus mykiss) hepatocytes. There was a direct relationship between hepatocyte glycogen content and total glycogen synthase, total glycogen phosphorylase and glycogen phosphorylase a activities, whereas the relationship was inverse between glycogen content and % glycogen synthase a and glycogen synthase a/glycogen phosphorylase a ratio. Incubation of hepatocytes with glucose (3 or 10 mmol·1-1) did not modify either glycogen synthase or glycogen phosphorylase activities. Insulin (porcine, 10-8 mol·1-1) in the medium significantly decreased total glycogen phosphorylase and glycogen phosphorylase a activities, but had no significant effect on glycogen synthase activities when compared to the controls (absence of insulin). In the presence of 10 mmol·1-1 glucose, insulin increased % glycogen synthase a and decreased % glycogen phosphorylase a activities in trout hepatocytes. Also, the effect of insulin on the activities of % glycogen synthase a and glycogen synthase a/glycogen phosphorylase a ratio were more pronounced at low than at high hepatocyte glycogen content. The results indicate that in trout hepatocytes both the glycogen synthetic and breakdown pathways are active concurrently in vitro and any subtle alterations in the phosphorylase to synthase ratio may determine the hepatic glycogen content. Insulin plays an important role in the regulation of glycogen metabolism in rainbow trout hepatocytes. The effect of insulin on hepatocyte glycogen content may be under the control of several factors, including plasma glucose concentration and hepatocyte glycogen content.

Key words

Glycogen Hepatocyte Insulin 13C NMR Rainbow trout, Oncorhynchus mykiss 

Abbreviations

13C NMR

13C nuclear magnetic resonance spectroscopy

GPase

glycogen phosphorylase

GSase

glycogen synthase

LDH

lactate dehydrogenase

PCA

perchloric acid

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References

  1. Borreback B, Waagbo R, Christophersen B, Tranulis M, Hemre G-I (1993) Adaptable hexokinase with low affinity for glucose in the liver of Atlantic salmon (Salmo salar). Comp Biochem Physiol 106B: 833–836Google Scholar
  2. Buddington RK, Hilton JW (1987) Intestinal adaptations of rainbow trout to changes in dietary carbohydrate. Am J Physiol 253: G489-G496PubMedGoogle Scholar
  3. Canioni P, Alger JR, Shulman RG (1983) Natural abundance 13C spectroscopy of liver and adipose tissue of the living rat. Biochemistry 22: 4974–4980CrossRefPubMedGoogle Scholar
  4. Cohen SM (1984) Application of 13C and 31P NMR to the study of hepatic metabolism. Fed Proc 43: 2657–2662PubMedGoogle Scholar
  5. Cowey CB, Knox D, Walton MJ, Adron JW (1977) The regulation of gluconcogenesis by diet and insulin in rainbow trout (Salmo gairdneri). Br J Nutr 38: 463–470PubMedGoogle Scholar
  6. Ciudad CJ, Carabaza A, Guinovart JJ (1986) Glucose 6-phosphate plays a central role in the activation of glycogen synthase by glucose in hepatocytes. Biochem Biophys Res Comm 141: 1195–1200CrossRefPubMedGoogle Scholar
  7. Foster GD, Moon TW (1987) Metabolism in sea raven (Hemitripterus americanus) hepatocytes: the effects of insulin and glucagon. Gen Comp Endocrinol 66: 102–115CrossRefPubMedGoogle Scholar
  8. Foster GD, Moon TW (1989) Insulin and the regulation of glycogen metabolism and gluconeogenesis in American eel hepatocytes. Gen Comp Endocrinol 73: 374–381PubMedGoogle Scholar
  9. Foster GD, Moon TW (1990) The role of glycogen phosphorylase in the regulation of glycogenolysis by insulin and glucagon in isolated eel (Anguilla rostrata) hepatocytes. Fish Physiol Biochem 8: 299–309Google Scholar
  10. Hemas DA, Whitton PD (1980) Control of hepatic glycogenolysis. Physiol Rev 60: 1–50Google Scholar
  11. Hers DA (1976) The control of glycogen metabolism in the liver. Annu Rev Biochem 45:167–189CrossRefPubMedGoogle Scholar
  12. Hilton JW, Atkinson JL (1982) Response of rainbow trout (Salmo gairdneri) to increased levels of available carbohydrate in practical trout diets. Br J Nutr 47: 597–607CrossRefPubMedGoogle Scholar
  13. Mommsen TP (1986) Comparative gluconcogenesis in hepatocytes from salmonid fishes. Can J Zool 64: 1110–1115Google Scholar
  14. Mommsen TP, Andrews PC, Plisetskaya EM (1987) Glucagon-like peptides activate hepatic gluconeogenesis. FEBS Lett 219: 227–232CrossRefPubMedGoogle Scholar
  15. Mommsen TP, Plisetskaya EM (1991) Insulin in fish and agnathans: history, structure and metabolic regulation. Rev Aquat Sci 4: 225–259Google Scholar
  16. Moon TW, Walsh PJ, Mommsen TP (1985) Fish hepatocytes: a model metabolic system. Can J Fish Aquat Sci 42: 1772–1782Google Scholar
  17. Moon TW, Foster GD, Plisetskaya EM (1989) Changes in peptide hormones and liver enzymes in the rainbow trout deprived of food for 6 weeks. Can J Zool 67: 2189–2193Google Scholar
  18. Mulmed LN, Gannon, MC, Gilboe DP, Tan AWH, Nuttall FQ (1979) Glycogen synthase, synthase phosphatase, and phosphorylase response to glucose in somatostatin-pretreated intact rats. Diabetes 28: 231–236PubMedGoogle Scholar
  19. Nutall FQ, Gannon MC (1993) Allosteric regulation of glycogen synthase in liver: a physiological dilemma. J Biol Chem 268: 13286–13290Google Scholar
  20. Nyfeler F, Fasel P, Walter P (1981) Short-term stimulation of net glycogen production by insulin in rat hepatocytes. Biochim Biophys Acta 675: 17–23PubMedGoogle Scholar
  21. Passoneau JV, Rottenberg DA (1973) An assessment of methods for measurement of glycogen synthetase activity including a new direct one-step assay. Anal Biochem 51: 528–541Google Scholar
  22. Pereira C, Vijayan MM, Moon TW (1995) In vitro metabolism of alanine and glucose and the response to insulin in fed and fasted rainbow trout. J Exp Zool (in press)Google Scholar
  23. Perry SF, Walsh PJ, Mommsen TP, Moon TW (1988) Metabolic consequences of hypercapnia in the rainbow trout, Salmo gairdneri: β-adrenergic effects. Gen Comp Endocrinol 69: 439–447CrossRefPubMedGoogle Scholar
  24. Plisetskaya EM, Bhattacharya S, Dickoff WW, Gorbman A (1984) The effect of insulin on amino acid metabolism and glycogen content in isolated liver cells of juvenile coho salmon (Oncorhynchus kisutch). Comp Biochem Physiol 78A: 773–778Google Scholar
  25. Rognstad R (1991) Possible futile glycogen cycling in hepatocytes. Biochem Arch 7: 221–226Google Scholar
  26. Scapin S, Di Giuseppe G (1994) Seasonal variations of glycogen synthase and phosphorylase activities in the liver of the frog Rana esculenta. Comp Biochem Physiol 107B: 189–195Google Scholar
  27. Shulman GI, Landau BR (1992) Pathways of glycogen repletion. Physiol Rev 72: 1019–1035PubMedGoogle Scholar
  28. Shulman GI, Rothman DL, Smith D, Johnson CM, Blair JB, Shulman RG, Defronzo RA (1985) Mechanism of liver glycogen repletion in vivo by nuclear magnetic resonance spectroscopy. J Clin Invest 76: 1229–1236PubMedGoogle Scholar
  29. Sillerud LO, Shulman RG (1983) Structure and metabolism of mammalian liver glycogen monitored by 13C NMR. Biochemistry 22: 1087–1094CrossRefPubMedGoogle Scholar
  30. Spence JT, Koudelka AP (1985) Pathway of glycogen synthesis from glucose in hepatocytes maintained in primary culture. J Biol Chem 260: 1521–1526PubMedGoogle Scholar
  31. Stalmans W, De Wulf H, Hers HG (1971) The effects of glucose and of a treatment by glucocorticoids on the inactivation in vitro of liver glycogen phosphorylase. Eur J Biochem 15: 9–12Google Scholar
  32. Stalmans W, de Wulf H, Hers HG (1971) The control of liver glycogen synthase phosphatase by phosphorylase. Eur J Biochem 18: 51–97CrossRefGoogle Scholar
  33. Stalmans W, De Wulf H, Hue L, Hers HG (1974a) The sequential inactivation of glycogen synthetase in the liver after the administration of glucose to mice and rats: the mechanism of the hepatic threshold to glucose. Eur J Biochem 41: 127–134CrossRefGoogle Scholar
  34. Stalmans W, Laloux M, Hers HG (1974b) The inactivation of liver phosphorylase a with glucose and AMP. Eur J Biochem 49: 415–427CrossRefPubMedGoogle Scholar
  35. Thomas AP, Martin-Roquero A, Williamson JR (1985) Interactions between insulin and α1 agents in the regulation of glycogen metabolism in isolated hepatocytes. J Biol Chem 260: 5963–5973PubMedGoogle Scholar
  36. Van de Werve G, Jeanrenaud B (1987) Liver glycogen metabolism: an overview. Diabetes/Metabolism Reviews 3: 47–78PubMedGoogle Scholar
  37. Van de Werve G, Stalmans W, Hers HG (1977) The effect of insulin on the glycogenolytic cascade and on the activity of glycogen synthase in the liver of anaesthetized rabbits. Biochem J 162: 143–146PubMedGoogle Scholar
  38. Vijayan MM, Foster GD, Moon TW (1993a) Effects of cortisol on hepatic carbohydrate metabolism and responsiveness to hormones in the sea raven, Hemitripterus americanus. Fish Physiol Biochem 12: 327–335Google Scholar
  39. Vijayan MM, Maule AG, Schreck CB, Moon TW (1993b) Hormonal control of hepatic glycogen metabolism in food-deprived, continously swimming coho salmon (Oncorhynchus kisutch). Can J Fish Aquat Sci 50: 1676–1682Google Scholar
  40. Vijayan MM, Moon TW (1992) Acute handling stress alters hepatic glycogen metabolism in food-deprived rainbow trout (Oncorhynchus mykiss). Can J Fish Aquat Sci 49: 2260–2266Google Scholar
  41. Vijayan MM, Pereira C, Moon TW (1994a) Hormonal stimulation of hepatocyte metabolism in rainbow trout following an acute handling stress. Comp Biochem Physiol 108C: 321–329Google Scholar
  42. Vijayan MM, Reddy PK, Leatherland JF, Moon TW (1994b) The effects of cortisol on hepatocyte metabolism in rainbow trout: a study using the steroid analogue RU486. Gen Comp Endocrinol 96: 75–84CrossRefPubMedGoogle Scholar
  43. Witters LA, Avruch L (1978) Insulin regulation of hepatic glycogen synthase and phosphorylase. Biochemistry 17: 406–410CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • C. Pereira
    • 1
  • M. M. Vijayan
    • 1
  • K. B. Storey
    • 1
  • R. A. Jones
    • 1
  • T. W. Moon
    • 1
  1. 1.Ottawa-Carleton Institute of Biology, Department of BiologyUniversity of OttawaOttawaCanada

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