Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1988 Aug;82(2):691–699. doi: 10.1172/JCI113649

Divergent mechanisms for the insulin resistant and hyperresponsive glucose transport in adipose cells from fasted and refed rats. Alterations in both glucose transporter number and intrinsic activity.

B B Kahn 1, I A Simpson 1, S W Cushman 1
PMCID: PMC303565  PMID: 3403723

Abstract

The effects of fasting and refeeding on the glucose transport response to insulin in isolated rat adipose cells have been examined using 3-O-methylglucose transport in intact cells and cytochalasin B binding and Western blotting in subcellular membrane fractions. After a 72-h fast, basal glucose transport activity decreases slightly and insulin-stimulated activity decreases greater than 85%. Following 48 h of fasting, insulin-stimulated glucose transport activity is diminished from 3.9 +/- 0.5 to 1.3 +/- 0.3 fmol/cell per min (mean +/- SEM). Similarly, the concentrations of glucose transporters are reduced with fasting in both the plasma membranes from insulin-stimulated cells from 38 +/- 5 to 18 +/- 3 pmol/mg of membrane protein and the low density microsomes from basal cells from 68 +/- 8 to 34 +/- 9 pmol/mg of membrane protein. Ad lib. refeeding for 6 d after a 48-h fast results in up to twofold greater maximally insulin-stimulated glucose transport activity compared with the control level (7.1 +/- 0.4 vs. 4.5 +/- 0.2 fmol/cell per min), before returning to baseline at 10 d. However, the corresponding concentration of glucose transporters in the plasma membranes is restored only to the control level (45 +/- 5 vs. 50 +/- 5 pmol/mg of membrane protein). Although the concentration of glucose transporters in the low density microsomes of basal cells remains decreased, the total number is restored to the control level due to an increase in low density microsomal protein. Thus, the insulin-resistant glucose transport in adipose cells from fasted rats can be explained by a decreased translocation of glucose transporters to the plasma membrane due to a depleted intracellular pool. In contrast, the insulin hyperresponsive glucose transport observed with refeeding appears to result from (a) a restored translocation of glucose transporters to the plasma membrane from a repleted intracellular pool and (b) enhanced plasma membrane glucose transporter intrinsic activity.

Full text

PDF
691

Images in this article

696Image
on p.696
696Image
on p.696
696Image
on p.696

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avruch J., Wallach D. F. Preparation and properties of plasma membrane and endoplasmic reticulum fragments from isolated rat fat cells. Biochim Biophys Acta. 1971 Apr 13;233(2):334–347. doi: 10.1016/0005-2736(71)90331-2. [DOI] [PubMed] [Google Scholar]
  2. Baldwin J. M., Gorga J. C., Lienhard G. E. The monosaccharide transporter of the human erythrocyte. Transport activity upon reconstitution. J Biol Chem. 1981 Apr 25;256(8):3685–3689. [PubMed] [Google Scholar]
  3. Björntorp P., Enzi G., Karlsson M., Kral J., Larsson B., Sjöström L., Smith U. Effects of refeeding on adipocyte metabolism in the rat. Int J Obes. 1980;4(1):11–19. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Carter-Su C., Okamoto K. Inhibition of hexose transport in adipocytes by dexamethasone: role of protein synthesis. Am J Physiol. 1985 Feb;248(2 Pt 1):E215–E223. doi: 10.1152/ajpendo.1985.248.2.E215. [DOI] [PubMed] [Google Scholar]
  6. Cushman S. W., Salans L. B. Determinations of adipose cell size and number in suspensions of isolated rat and human adipose cells. J Lipid Res. 1978 Feb;19(2):269–273. [PubMed] [Google Scholar]
  7. Cushman S. W. Structure-function relationships in the adipose cell. I. Ultrastructure of the isolated adipose cell. J Cell Biol. 1970 Aug;46(2):326–341. doi: 10.1083/jcb.46.2.326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cushman S. W., Wardzala L. J. Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. Apparent translocation of intracellular transport systems to the plasma membrane. J Biol Chem. 1980 May 25;255(10):4758–4762. [PubMed] [Google Scholar]
  9. Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. II. Synthesis of constitutive microsomal enzymes in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):97–117. doi: 10.1083/jcb.30.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fleischer B. Isolation and characterization of Golgi apparatus and membranes from rat liver. Methods Enzymol. 1974;31:180–191. doi: 10.1016/0076-6879(74)31020-8. [DOI] [PubMed] [Google Scholar]
  11. Flier J. S., Mueckler M. M., Usher P., Lodish H. F. Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes. Science. 1987 Mar 20;235(4795):1492–1495. doi: 10.1126/science.3103217. [DOI] [PubMed] [Google Scholar]
  12. Guerre-Millo M., Lavau M., Horne J. S., Wardzala L. J. Proposed mechanism for increased insulin-mediated glucose transport in adipose cells from young, obese Zucker rats. Large intracellular pool of glucose transporters. J Biol Chem. 1985 Feb 25;260(4):2197–2201. [PubMed] [Google Scholar]
  13. HUBER A. M., GERSHOFF S. N., ANTONIADES H. N. THE EFFECT OF FASTING AND REFEEDING ON ISOLATED RAT ADIPOSE TISSUE ACTIVITY IN THE PRESENCE OF CRYSTALLINE AND "BOUND" INSULIN. Metabolism. 1965 May;14:619–624. doi: 10.1016/s0026-0495(65)80024-5. [DOI] [PubMed] [Google Scholar]
  14. Hirsch J., Gallian E. Methods for the determination of adipose cell size in man and animals. J Lipid Res. 1968 Jan;9(1):110–119. [PubMed] [Google Scholar]
  15. Hissin P. J., Foley J. E., Wardzala L. J., Karnieli E., Simpson I. A., Salans L. B., Cushman S. W. Mechanism of insulin-resistant glucose transport activity in the enlarged adipose cell of the aged, obese rat. J Clin Invest. 1982 Oct;70(4):780–790. doi: 10.1172/JCI110674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hissin P. J., Karnieli E., Simpson I. A., Salans L. B., Cushman S. W. A possible mechanism of insulin resistance in the rat adipose cell with high-fat/low-carbohydrate feeding. Depletion of intracellular glucose transport systems. Diabetes. 1982 Jul;31(7):589–592. doi: 10.2337/diab.31.7.589. [DOI] [PubMed] [Google Scholar]
  17. Horuk R., Rodbell M., Cushman S. W., Wardzala L. J. Proposed mechanism of insulin-resistant glucose transport in the isolated guinea pig adipocyte. Small intracellular pool of glucose transporters. J Biol Chem. 1983 Jun 25;258(12):7425–7429. [PubMed] [Google Scholar]
  18. Joost H. G., Weber T. M., Cushman S. W., Simpson I. A. Insulin-stimulated glucose transport in rat adipose cells. Modulation of transporter intrinsic activity by isoproterenol and adenosine. J Biol Chem. 1986 Aug 5;261(22):10033–10036. [PubMed] [Google Scholar]
  19. Kahn B. B., Cushman S. W. Mechanism for markedly hyperresponsive insulin-stimulated glucose transport activity in adipose cells from insulin-treated streptozotocin diabetic rats. Evidence for increased glucose transporter intrinsic activity. J Biol Chem. 1987 Apr 15;262(11):5118–5124. [PubMed] [Google Scholar]
  20. Karnieli E., Armoni M., Cohen P., Kanter Y., Rafaeloff R. Reversal of insulin resistance in diabetic rat adipocytes by insulin therapy. Restoration of pool of glucose transporters and enhancement of glucose-transport activity. Diabetes. 1987 Aug;36(8):925–931. doi: 10.2337/diab.36.8.925. [DOI] [PubMed] [Google Scholar]
  21. Karnieli E., Barzilai A., Rafaeloff R., Armoni M. Distribution of glucose transporters in membrane fractions isolated from human adipose cells. Relation to cell size. J Clin Invest. 1986 Oct;78(4):1051–1055. doi: 10.1172/JCI112660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Karnieli E., Hissin P. J., Simpson I. A., Salans L. B., Cushman S. W. A possible mechanism of insulin resistance in the rat adipose cell in streptozotocin-induced diabetes mellitus. Depletion of intracellular glucose transport systems. J Clin Invest. 1981 Sep;68(3):811–814. doi: 10.1172/JCI110318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Karnieli E., Zarnowski M. J., Hissin P. J., Simpson I. A., Salans L. B., Cushman S. W. Insulin-stimulated translocation of glucose transport systems in the isolated rat adipose cell. Time course, reversal, insulin concentration dependency, and relationship to glucose transport activity. J Biol Chem. 1981 May 25;256(10):4772–4777. [PubMed] [Google Scholar]
  24. Kasuga M., Akanuma Y., Iwamoto Y., Kosaka K. Effects of fasting and refeeding of insulin receptors and glucose metabolism in rat adipocytes. Endocrinology. 1977 May;100(5):1384–1390. doi: 10.1210/endo-100-5-1384. [DOI] [PubMed] [Google Scholar]
  25. Kono T., Suzuki K., Dansey L. E., Robinson F. W., Blevins T. L. Energy-dependent and protein synthesis-independent recycling of the insulin-sensitive glucose transport mechanism in fat cells. J Biol Chem. 1981 Jun 25;256(12):6400–6407. [PubMed] [Google Scholar]
  26. Kuroda M., Honnor R. C., Cushman S. W., Londos C., Simpson I. A. Regulation of insulin-stimulated glucose transport in the isolated rat adipocyte. cAMP-independent effects of lipolytic and antilipolytic agents. J Biol Chem. 1987 Jan 5;262(1):245–253. [PubMed] [Google Scholar]
  27. Lienhard G. E., Kim H. H., Ransome K. J., Gorga J. C. Immunological identification of an insulin-responsive glucose transporter. Biochem Biophys Res Commun. 1982 Apr 14;105(3):1150–1156. doi: 10.1016/0006-291x(82)91090-7. [DOI] [PubMed] [Google Scholar]
  28. MOORE R. O. INFLUENCE OF FASTING AND REFEEDING ON RESPONSE OF ADIPOSE TISSUE TO INSULIN. Am J Physiol. 1963 Aug;205:222–224. doi: 10.1152/ajplegacy.1963.205.2.222. [DOI] [PubMed] [Google Scholar]
  29. Mosher K. M., Young D. A., Munck A. Evidence for irreversible, actinomycin D-sensitive, and temperature-sensitive steps following binding of cortisol to glucocorticoid receptors and preceding effects on glucose metabolism in rat thymus cells. J Biol Chem. 1971 Feb 10;246(3):654–659. [PubMed] [Google Scholar]
  30. Olefsky J. M. Effects of fasting on insulin binding, glucose transport, and glucose oxidation in isolated rat adipocytes: relationships between insulin receptors and insulin action. J Clin Invest. 1976 Dec;58(6):1450–1460. doi: 10.1172/JCI108601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Owens J. L., Thompson D., Shah N., DiGirolamo M. Effects of fasting and refeeding in the rat on adipocyte metabolic functions and response to insulin. J Nutr. 1979 Sep;109(9):1584–1591. doi: 10.1093/jn/109.9.1584. [DOI] [PubMed] [Google Scholar]
  32. Pedersen O., Hjøllund E., Sørensen N. S. Insulin receptor binding and insulin action in human fat cells: effects of obesity and fasting. Metabolism. 1982 Sep;31(9):884–895. doi: 10.1016/0026-0495(82)90177-9. [DOI] [PubMed] [Google Scholar]
  33. RODBELL M. METABOLISM OF ISOLATED FAT CELLS. I. EFFECTS OF HORMONES ON GLUCOSE METABOLISM AND LIPOLYSIS. J Biol Chem. 1964 Feb;239:375–380. [PubMed] [Google Scholar]
  34. Simpson I. A., Sonne O. A simple, rapid, and sensitive method for measuring protein concentration in subcellular membrane fractions prepared by sucrose density ultracentrifugation. Anal Biochem. 1982 Jan 15;119(2):424–427. doi: 10.1016/0003-2697(82)90608-x. [DOI] [PubMed] [Google Scholar]
  35. Simpson I. A., Yver D. R., Hissin P. J., Wardzala L. J., Karnieli E., Salans L. B., Cushman S. W. Insulin-stimulated translocation of glucose transporters in the isolated rat adipose cells: characterization of subcellular fractions. Biochim Biophys Acta. 1983 Dec 19;763(4):393–407. doi: 10.1016/0167-4889(83)90101-5. [DOI] [PubMed] [Google Scholar]
  36. Smith M. M., Robinson F. W., Watanabe T., Kono T. Partial characterization of the glucose transport activity in the Golgi-rich fraction of fat cells. Biochim Biophys Acta. 1984 Aug 22;775(2):121–128. doi: 10.1016/0005-2736(84)90162-7. [DOI] [PubMed] [Google Scholar]
  37. Smith U., Kuroda M., Simpson I. A. Counter-regulation of insulin-stimulated glucose transport by catecholamines in the isolated rat adipose cell. J Biol Chem. 1984 Jul 25;259(14):8758–8763. [PubMed] [Google Scholar]
  38. Suzuki K., Kono T. 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. 1980 May;77(5):2542–2545. doi: 10.1073/pnas.77.5.2542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Timmers K., Knittle J. L. Effects of undernutrition and refeeding on enzyme activities and rates of glucose catabolism in rat epididymal adipose tissue. J Nutr. 1980 Jun;110(6):1176–1184. doi: 10.1093/jn/110.6.1176. [DOI] [PubMed] [Google Scholar]
  40. Vinten J., Petersen N., Sonne B., Galbo H. Effect of physical training on glucose transporters in fat cell fractions. Biochim Biophys Acta. 1985 Aug 16;841(2):223–227. doi: 10.1016/0304-4165(85)90025-x. [DOI] [PubMed] [Google Scholar]
  41. Wardzala L. J., Cushman S. W., Salans L. B. Mechanism of insulin action on glucose transport in the isolated rat adipose cell. Enhancement of the number of functional transport systems. J Biol Chem. 1978 Nov 25;253(22):8002–8005. [PubMed] [Google Scholar]
  42. Wheeler T. J., Simpson I. A., Sogin D. C., Hinkle P. C., Cushman S. W. Detection of the rat adipose cell glucose transporter with antibody against the human red cell glucose transporter. Biochem Biophys Res Commun. 1982 Mar 15;105(1):89–95. doi: 10.1016/s0006-291x(82)80014-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES