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Insulin-Stimulated Glucose Uptake Involves the Transition of Glucose Transporters to a Caveolae-Rich Fraction within the Plasma Membrane: Implications for Type II Diabetes

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Abstract

Background

Adipose and muscle tissues express an insulin-sensitive glucose transporter (GLUT4). This transporter has been shown to translocate from intracellular stores to the plasma membrane following insulin stimulation. The molecular mechanisms signalling this event and the details of the translocation pathway remain unknown. In type II diabetes, the cellular transport of glucose in response to insulin is impaired, partly explaining why blood-glucose levels in patients are not lowered by insulin as in normal individuals.

Materials and Methods

Isolated rat epididymal adipocytes were stimulated with insulin and subjected to subcellular fractionation and to measurement of glucose uptake. A caveolae-rich fraction was isolated from the plasma membranes after detergent solubilization and ultracentrifugal floatation in a sucrose gradient. Presence of GLUT4 and caveolin was determined by immunoblotting after SDS-PAGE.

Results

In freshly isolated adipocytes, insulin induced a rapid translocation of GLUT4 to the plasma membrane fraction, which was followed by a slower transition of the transporter into a detergent resistant caveolae-rich region of the plasma membrane. The insulin-stimulated appearance of transporters in the caveolae-rich fraction occurred in parallel with enhanced glucose uptake by cells. Treatment with isoproterenol plus adenosine deaminase rapidly inhibited insulin-stimulated glucose transport by 40%, and at the same time GLUT4 disappeared from the caveolae-rich fraction and from plasma membranes as a whole.

Conclusions

Insulin stimulates glucose uptake in adipocytes by rapidly translocating GLUT4 from intracellular stores to the plasma membrane. This is followed by a slower transition of GLUT4 to the caveolae-rich regions of the plasma membrane, where glucose transport appears to take place. These results have implications for an understanding of the defect in glucose transport involved in type II diabetes.

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References

  1. Cushman SW, Wardzala LJ. (1980) Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. J. Biol Chem. 255: 4758–4762.

    PubMed  CAS  Google Scholar 

  2. Suzuki K, Kono T. (1980) Evidence that insulin causes translocation of glucose transport activity to the plasma membrane from an intracellular storage site. Proc. Natl Acad. Sci U.S.A. 77: 2542–2545.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Karnieli E, Zarnowski M, Hissin PJ, Simpson IA, Salans LB, Cushman SW. (1981) Insulinstimulated translocation of glucose transport systems in the isolated rat adipose cell. J. Biol. Chem. 256: 4772–4777.

    PubMed  CAS  Google Scholar 

  4. Satoh S, Nishimura H, Clark A, et al. (1993) Use of bis-mannitol photolabel to elucidate insulin-regulated GLUT-4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action. J. Biol Chem. 268: 17820–17829.

    PubMed  CAS  Google Scholar 

  5. Gibbs EM, Lienhard GE, Gould GW. (1988) Insulin-induced translocation of glucose transporters to the plasma membrane precedes full stimulation of hexose transport. Biochemistry 27: 6681–6685.

    Article  CAS  PubMed  Google Scholar 

  6. Joost HG, Weber TM, Cushman SW. (1988) Qualitative and quantitative comparison of glucose transport activity and glucose transporter concentration in plasma membranes from basal and insulin-stimulated rat adipose cells. Biochem. J. 249: 155–161.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Lange K, Brandt U. (1990) Insulin-responsive glucose transporters are concentrated in a cell surface-derived membrane fraction of 3T3-L1 adipocytes. FEBS Lett. 261: 459–463.

    Article  CAS  PubMed  Google Scholar 

  8. Clark AE, Holman GD, Kozka IJ. (1991) Determination of the rates of appearance and loss of glucose transporters at the cell surface of rat adipose cells. Biochem. J. 278: 235–241.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Smith RM, Charron MJ, Shah N, Lodish HF, Jarett L. (1991) Immunoelectron microscopic demonstration of insulin-stimulated translocation of glucose transporters to the plasma membrane of isolated rat adipocytes and masking of the carboxyl-terminal epitope of intracellular GLUT4. Proc. Natl. Acad. sci. U.S.A. 88: 6893–6897.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Vannucci SJ, Nishimura H, Satoh S, Cushman SW, Holman GD, Simpson IA. (1992) Cell surface accessibility of GLUT4 glucose transporters in insulin-stimulated rat adipose cells, modulation by isoprenaline and adenosine. Biochem. J. 288: 325–330.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Holman GD, Leggio LL, Cushman SW. (1994) Insulin-stimulated GLUT4 glucose transporter recycling. J. Biol. Chem. 269: 17516–17524.

    PubMed  CAS  Google Scholar 

  12. Scherer PE, Lisanti MP, Baldini G, Sargiacomo M, Mastick CC, Lodish HF. (1994) Induction of caveolin during adipogenesis and association of GLUT4 with cavolin-rich vesicles. J. Cell Biol. 127: 1233–1243.

    Article  CAS  PubMed  Google Scholar 

  13. Kandror KV, Stephens JM, Pilch PF. (1995) Expression and compartmentalisation of caveolin in adipose cells: co-ordinate regulation with and structural segregation from GLUT4. J. Cell Biol 129: 999–1006.

    Article  CAS  PubMed  Google Scholar 

  14. Parpal S, Gustavsson J, Strâlfors P. (1995) Isolation of phosphooligosaccharide/phosphoinositol glycan from caveolae and from cytosol of insulin-stimulated cells. J. Cell Biol. 131: 125–135.

    Article  CAS  PubMed  Google Scholar 

  15. Strålfors P, Honnor RC. (1989) Insulin-induced dephosphorylation of hormone-sensitive lipase, correlation with lipolysis and cAMP-dependent protein kinase activity. Eur. J. Biochem. 182: 379–385.

    Article  PubMed  Google Scholar 

  16. Strålfors P. (1988) Insulin stimulation of glucose uptake can be mediated by diacylglycerol in adipocytes. Nature (Lond.) 335: 554–556.

    Article  Google Scholar 

  17. Oka Y, Czech M. (1984) Photoaffinity labelling of insulin-sensitive hexose transporters in intact rat adipocytes, direct evidence that latent transporters become exposed to the extracellular space in response to insulin. J. Biol. Chem. 259: 8125–8133.

    PubMed  CAS  Google Scholar 

  18. McKeel DW, Jarett L. (1970) Preparation and characterisation of a plasma membrane fraction from isolated fat cells. J. Cell Biol. 44: 417–432.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Chang WJ, Ying YS, Rothberg KG, et al. (1994) Purification and characterisation of smooth muscle cell caveolae. J. Cell Biol. 126: 127–138.

    Article  CAS  PubMed  Google Scholar 

  20. Sargiacomo M, Sudol M, Tang ZL, Lisanti MP. (1993) Signal transducing molecules and glycosyl-phosphatidylinositol-linked proteins form a caveolin-rich insoluble complex in MDCK cells. J. Cell Biol. 122: 789–807.

    Article  CAS  PubMed  Google Scholar 

  21. Parton GP, Simons K. (1995) Digging into caveolae. Science 269: 1398–1399.

    Article  CAS  PubMed  Google Scholar 

  22. Smart EJ, Ying YS, Mineo C, Anderson RGW. (1995) A detergent-free method for purifying caveolae membrane from tissue culture cells. Proc. Natl. Acad. sci. U.S.A. 92: 10104–10108.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Department of Cell Biology, Faculty of Health Sciences, University of Linköping, S-58185, Linköping, Sweden

    Johanna Gustavsson, Santiago Parpal & Peter Strålfors

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  1. Johanna Gustavsson
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  2. Santiago Parpal
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  3. Peter Strålfors
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Gustavsson, J., Parpal, S. & Strålfors, P. Insulin-Stimulated Glucose Uptake Involves the Transition of Glucose Transporters to a Caveolae-Rich Fraction within the Plasma Membrane: Implications for Type II Diabetes. Mol Med 2, 367–372 (1996). https://doi.org/10.1007/BF03401634

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