Skip to main content
Log in

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

Journal of Comparative Physiology B Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Abbreviations

13C NMR:

13C nuclear magnetic resonance spectroscopy

GPase:

glycogen phosphorylase

GSase:

glycogen synthase

LDH:

lactate dehydrogenase

PCA:

perchloric acid

References

  • 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: 833–836

    Google Scholar 

  • Buddington RK, Hilton JW (1987) Intestinal adaptations of rainbow trout to changes in dietary carbohydrate. Am J Physiol 253: G489-G496

    CAS  PubMed  Google Scholar 

  • Canioni P, Alger JR, Shulman RG (1983) Natural abundance 13C spectroscopy of liver and adipose tissue of the living rat. Biochemistry 22: 4974–4980

    Article  CAS  PubMed  Google Scholar 

  • Cohen SM (1984) Application of 13C and 31P NMR to the study of hepatic metabolism. Fed Proc 43: 2657–2662

    CAS  PubMed  Google Scholar 

  • 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: 463–470

    CAS  PubMed  Google Scholar 

  • 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: 1195–1200

    Article  CAS  PubMed  Google Scholar 

  • Foster GD, Moon TW (1987) Metabolism in sea raven (Hemitripterus americanus) hepatocytes: the effects of insulin and glucagon. Gen Comp Endocrinol 66: 102–115

    Article  CAS  PubMed  Google Scholar 

  • Foster GD, Moon TW (1989) Insulin and the regulation of glycogen metabolism and gluconeogenesis in American eel hepatocytes. Gen Comp Endocrinol 73: 374–381

    CAS  PubMed  Google Scholar 

  • 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: 299–309

    CAS  Google Scholar 

  • Hemas DA, Whitton PD (1980) Control of hepatic glycogenolysis. Physiol Rev 60: 1–50

    Google Scholar 

  • Hers DA (1976) The control of glycogen metabolism in the liver. Annu Rev Biochem 45:167–189

    Article  CAS  PubMed  Google Scholar 

  • 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: 597–607

    Article  CAS  PubMed  Google Scholar 

  • Mommsen TP (1986) Comparative gluconcogenesis in hepatocytes from salmonid fishes. Can J Zool 64: 1110–1115

    CAS  Google Scholar 

  • Mommsen TP, Andrews PC, Plisetskaya EM (1987) Glucagon-like peptides activate hepatic gluconeogenesis. FEBS Lett 219: 227–232

    Article  CAS  PubMed  Google Scholar 

  • Mommsen TP, Plisetskaya EM (1991) Insulin in fish and agnathans: history, structure and metabolic regulation. Rev Aquat Sci 4: 225–259

    CAS  Google Scholar 

  • Moon TW, Walsh PJ, Mommsen TP (1985) Fish hepatocytes: a model metabolic system. Can J Fish Aquat Sci 42: 1772–1782

    CAS  Google Scholar 

  • 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: 2189–2193

    CAS  Google Scholar 

  • 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: 231–236

    CAS  PubMed  Google Scholar 

  • Nutall FQ, Gannon MC (1993) Allosteric regulation of glycogen synthase in liver: a physiological dilemma. J Biol Chem 268: 13286–13290

    Google Scholar 

  • Nyfeler F, Fasel P, Walter P (1981) Short-term stimulation of net glycogen production by insulin in rat hepatocytes. Biochim Biophys Acta 675: 17–23

    CAS  PubMed  Google Scholar 

  • 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: 528–541

    Google Scholar 

  • 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)

  • 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: 439–447

    Article  CAS  PubMed  Google Scholar 

  • 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: 773–778

    CAS  Google Scholar 

  • Rognstad R (1991) Possible futile glycogen cycling in hepatocytes. Biochem Arch 7: 221–226

    CAS  Google Scholar 

  • 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: 189–195

    Google Scholar 

  • Shulman GI, Landau BR (1992) Pathways of glycogen repletion. Physiol Rev 72: 1019–1035

    CAS  PubMed  Google Scholar 

  • 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: 1229–1236

    CAS  PubMed  Google Scholar 

  • Sillerud LO, Shulman RG (1983) Structure and metabolism of mammalian liver glycogen monitored by 13C NMR. Biochemistry 22: 1087–1094

    Article  CAS  PubMed  Google Scholar 

  • Spence JT, Koudelka AP (1985) Pathway of glycogen synthesis from glucose in hepatocytes maintained in primary culture. J Biol Chem 260: 1521–1526

    CAS  PubMed  Google Scholar 

  • 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: 9–12

    Google Scholar 

  • Stalmans W, de Wulf H, Hers HG (1971) The control of liver glycogen synthase phosphatase by phosphorylase. Eur J Biochem 18: 51–97

    Article  Google Scholar 

  • 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: 127–134

    Article  CAS  Google Scholar 

  • Stalmans W, Laloux M, Hers HG (1974b) The inactivation of liver phosphorylase a with glucose and AMP. Eur J Biochem 49: 415–427

    Article  CAS  PubMed  Google Scholar 

  • 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: 5963–5973

    CAS  PubMed  Google Scholar 

  • Van de Werve G, Jeanrenaud B (1987) Liver glycogen metabolism: an overview. Diabetes/Metabolism Reviews 3: 47–78

    PubMed  Google Scholar 

  • 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: 143–146

    PubMed  Google Scholar 

  • 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: 327–335

    CAS  Google Scholar 

  • 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: 1676–1682

    CAS  Google Scholar 

  • 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: 2260–2266

    CAS  Google Scholar 

  • Vijayan MM, Pereira C, Moon TW (1994a) Hormonal stimulation of hepatocyte metabolism in rainbow trout following an acute handling stress. Comp Biochem Physiol 108C: 321–329

    Google Scholar 

  • 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: 75–84

    Article  CAS  PubMed  Google Scholar 

  • Witters LA, Avruch L (1978) Insulin regulation of hepatic glycogen synthase and phosphorylase. Biochemistry 17: 406–410

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by L.C.-H. Wang

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pereira, C., Vijayan, M.M., Storey, K.B. et al. Role of glucose and insulin in regulating glycogen synthase and phosphorylase activities in rainbow trout hepatocytes. J Comp Physiol B 165, 62–70 (1995). https://doi.org/10.1007/BF00264687

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00264687

Key words

Navigation