Summary
Observations in vivo suggest that insulin acts as a long-term regulator of hexose uptake in fat cells. In the present study, we examined the long-term effect of insulin on hexose uptake in vitro. Exposure of fully differentiated mouse 3T3-L1 adipocytes to insulin induced a time-, concentration-, and protein synthesis-dependent increase in basal 2-deoxyglucose uptake (up to 40%) and a decrease in the ‘acute’ insulin response. The decrease in insulin effect was due to post-receptor alterations, since insulin binding was not substantially altered. The increase in basal 2-deoxyglucose uptake was due to an increase in the apparent Vmax of the transport system rather than to the observed increase (30%) in hexokinase activity, since the concentration of non-phosphorylated 2-deoxyglucose inside the cell was far below the extracellular concentration. The increase in apparent Vmax was most likely due to a protein synthesis-dependent increase in de novo synthesis of hexose transporters. Glucose was not essential for the effect. The mechanism responsible for the loss in insulin response remains to be solved. It can be concluded that insulin has the ability to act as a long-term regulator of hexose uptake in fat cells in vitro.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
DeFronzo RA, Hendler R, Simonson D (1982) Insulin resistance is a prominent factor of insulin-dependent diabetes. Diabetes 31: 795–801
Pedersen O, Hjøllund E (1982) Insulin receptor binding to fat and blood cells and insulin action in fat cells from insulin dependent diabetics. Diabetcs 31: 706–715
Wieringa Tj, Krans HMJ (1978) Reduced glucose transport and increased binding of insulin in adipocytes from diabetic and fasted rats. Biochim Biophys Acta 538: 563–570
Radder JK, Wieringa Tj, Bos MP, Krans HMJ (1984) The effect of normalization of blood glucose levels in streptozotocin-diabetic rats with continuous intravenous insulin infusion on insulin binding and glucose transport of isolated adipocytes. In: Shafrir E, Renold AE (eds) Lessons from animal diabetes. John Libbey, London, 622–626
Kasuga M, Akanuma Y, Iwamoto Y, Kosaka K (1978) Insulin binding and glucose metabolism in adipocytes of streptozotocin diabetic rats. Am J Physiol 235: E175-E183
Kobayashi M, Olefsky JM (1979) Effects of streptozotocin induced diabetes on insulin binding, glucose transport and intracellular glucose metabolism in isolated rat adipocytes. Diabetes 28: 87–95
Olefsky JM (1976) Effects of fasting on insulin binding, glucose transport and glucose oxidation in isolated rat adipocytes. J Clin Invest 58: 1450–1460
Oka Y, Akanuma Y, Kasuga M, Kosaka K (1980) Effect of a high glucose diet on insulin binding and insulin action in rat adipocytes. Diabetologia 19: 468–474
Saekow M, Olefsky JM (1980) Effect of a high carbohydrate diet on adipocyte glucose metabolism in spontaneously obese rats and insulin deficient diabetic rats. Endocrinology 107: 2004–2010
Ip C, Tepperman HM, deWitt J, Tepperman J (1977) The effect of diet fat on rat adipocyte glucose transport. Horm Metab Res 9: 218–222
Kobayashi M, Olefsky JM (1978) Long-term regulation of adipocyte glucose transport capacity by circulating insulin in rats. J Clin Invest 62: 73–81
Livingston JN, Purvis BJ, Lockwood DH (1978) Insulin-induced changes in insulin binding and insulin sensitivity of adipocytes. Metabolism 27: 2009–2014
Livingston JN, Purvis BJ, Lockwood DH (1978) Insulin-dependent regulation of insulin sensitivity of adipocytes. Nature 273: 394–396
Marshall S, Olefsky JM (1980) Effects of insulin incubation on insulin binding, glucose transport and insulin degradation by isolated rat adipocytes. J Clin Invest 66: 763–772
Vega FV, Kono T (1979) Sugar transport in fat cells: effect of mechanical agitation, cell-bound insulin and temperature. Arch Biochem Biophys 192: 120–127
Kikuchi K, Schwartz C, Creacy S, Larner J (1981) Independent control of selected insulin sensitive cell membrane and intracellular functions — the linkage of cell membrane and intracellular events controlled by insulin. Mol Cell Biochem 37: 125–130
El-Allaway RMM, Gliemann J (1972) Trypsin treatment of adipocytes: effect on sensitivity to insulin. Biochim Biophys Acta 271: 97–109
Green H, Kehinde O (1974) Sublines of mouse 3T3 cells that accumulate lipid. Cell 1: 113–116
Green H (1979) Adipose conversion: a program of differentiation. In: Ailhaud G (ed) Obesity, cellular and molecular aspects. Inserm, Nice 87: 15–24
Rubin CS, Hirsch A, Fung C, Rosen OM (1978) Development of hormone receptors and hormonal responsiveness in vitro. J Biol Chem 253: 7570–7578
Reed BC, Lane MD (1980) Insulin receptor synthesis and turnover in differentiating 3T3-L1 preadipocytes. Proc Natl Acad Sci USA 77: 285–289
Sedmak JJ, Grossberg SE (1977) A rapid, sensitive and versatile assay for protein using Coomassie Brilliant Blue G 250. Anal Biochem 79: 544–553
Wieringa Tj, van Putten JPM, Krans HMJ (1981) Rapid phloretin induced dephosphorylation of 2-deoxyglucose 6-phosphate in rat adipocytes. Biochem Biophys Res Commun 103: 841–847
Gliemann J, Osterlind K, Vinten J, Gammeltoft S (1972) A procedure for measurement of distribution spaces in isolated fat cells. Biochim Biophys Acta 286: 1–9
Freychet P, Kahn CR, Roth J, Neville DM Jr (1972) Insulin interactions with liver plasma membrane. J Biol Chem 247: 3953–3961
Resh MD (1982) Development of insulin responsiveness of the glucose transporter and the (Na+, K+)-adenosine triphosphatase during in vitro adipocyte differentiation. J Biol Chem 257: 6978–6986
Karlsson FA, Grunfeld C, Kahn CR, Roth J (1979) Regulation of insulin receptors and insulin responsiveness in 3T3-L1 fatty fibroblasts. Endocrinology 104: 1383–1392
Rosen OM, Smith CJ, Fung C, Rubin CS (1979) Development of hormone receptors and hormone responsiveness in vitro. J Biol Chem 253: 7579–7583
Reed BC, Ronnett GV, Clements PR, Lane MD (1981) Regulation of insulin receptor metabolism. J Biol Chem 256: 3917–3925
Ronnett GV, Knutson VP, Lane MD (1982) Insulin-induced down-regulation in 3T3-L1 adipocytes. J Biol Chem 257: 4285–4291
Ronnett GV, Tennekoon G, Knutson VP, Lane MD (1983) Kinetics of insulin receptor transit and removal from the plasma membrane. J Biol Chem 258: 283–290
Chang T, Polakis SE (1978) Differentiation of 3T3-L1 fibroblasts to adipocytes. J Biol Chem 253: 4693–4696
Kahn CR, Baird K, Flier JS, Jarrett DB (1977) Effects of autoantibodies to the insulin receptor on isolated adipocytes. J Clin Invest 60: 1094–1106
Karlsson FA, van Obberghen E, Grunfeld C, Kahn CR (1979) Desensitization of the insulin receptor at an early post receptor step by prolonged exposure to anti-receptor antibody. Proc Natl Acad Sci USA 76: 809–813
Foley JE, Foley R, Gliemann J (1980) Rate-limiting steps of 2-deoxyglucose uptake in rat adipocytes. Biochim Biophys Acta 599: 689–698
de Jonge PC, Wieringa Tj, van Putten JPM, Krans HMJ, van Dam K (1983) Phloretin — an uncoupler and inhibitor of mitochondrial oxidative phosphorylation. Biochim Biophys Acta 722: 219–225
Ciaraldi TP, Kolterman OG, Scarlett JA, Kao M, Olefsky JM (1982) Role of glucose transport in postreceptor defect in non-insulin-dependent diabetes mellitus. Diabetes 31: 1016–1022
Scarlett JA, Kolterman OG, Ciaraldi TP, Kao M, Olefsky JM (1983) Insulin treatment reverses the postreceptor defect in adipocyte 3-O-methylglucose transport in type II diabetes mellitus. J Clin Endocrinol Metab 56: 1195–1201
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van Putten, J.P.M., Wieringa, T. & Krans, H.M.J. Long-term regulation of hexose transport by insulin in cultured mouse (3T3) adipocytes. Diabetologia 28, 51–56 (1985). https://doi.org/10.1007/BF00277000
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00277000