Summary
Glucose is the main metabolic fuel in mammalian cells. Glucose entry into cells is facilitated by a family of ubiquitously expressed glucose transporter proteins. Typically, glucose transporters are localized on the plasma membrane. One notable exception is the glucose transporter isoform 4 (Glut4), which is specifically expressed in insulin sensitive tissues, i.e., skeletal muscle, heart muscle, and fat, and is responsible for the insulin effect on blood glucose clearance (1). Under basal conditions, Glut4 is compartmentalized in intracellular membrane vesicles and thus has no access to the extracellular space. Upon insulin administration, Glut4-containing vesicles fuse with the plasma membrane and deliver the transporter to its site of action. As a result, Glut4 content on the plasma membrane is increased, and glucose uptake in the cell is significantly elevated. Here, we describe two complementary techniques. The first one uses tritiated 2-deoxyglucose and is designed to measure insulin-stimulated glucose transport into cultured adipose cells. The second allows one to quantify the degree of Glut4 translocation from an intracellular compartment to the plasma membrane.
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Saltiel AR and Kahn CR (2001) Insulin signalling and the regulation of glucose and lipid metabolism. Nature 414:799–806
Kandror KV (2003) A long search for Glut4 activation. Sci STKE 2003:PE5
Shi J, Kandror KV (2005) Sortilin is essential and sufficient for the formation of Glut4-stor-age vesicles in 3T3-L1 adipocytes. Dev Cell 9:99–108
Carvalho E, Schellhorn SE, Zabolotny JM, Martin S, Tozzo E, Peroni OD, Houseknecht KL, Mundt A, James DE, Kahn BB (2004) GLUT4 overexpression or deficiency in adipocytes of transgenic mice alters the composition of GLUT4 vesicles and the subcellular localization of GLUT4 and insulin-responsive aminopeptidase. J Biol Chem 279:21598–21605
Kanai F, Nishioka Y, Hayashi H, Kamohara S, Todaka M, Ebina Y (1993) Direct demonstration of insulin-induced GLUT4 translocation to the surface of intact cells by insertion of a c-myc epitope into an exofacial GLUT4 domain. J Biol Chem 268:14523–14526
Kishi K, Muromoto N, Nakaya Y, Miyata I, Hagi A, Hayashi H, Ebina Y (1998) Bradykinin directly triggers GLUT4 translocation via an insulin-independent pathway. Diabetes 47:550–558
Ueyama A, Yaworsky KL, Wang Q, Ebina Y, Klip A (1999) GLUT-4 myc ectopic expression in L6 myoblasts generates a GLUT-4-specific pool conferring insulin sensitivity. Am J Physiol 277:E572–E578
Karylowski O, Zeigerer A, Cohen A, McGraw TE (2004) GLUT4 is retained by an intracel-lular cycle of vesicle formation and fusion with endosomes. Mol Biol Cell 15:870–882
Govers R, Coster AC, James DE (2004) Insulin increases cell surface GLUT4 levels by dose dependently discharging GLUT4 into a cell surface recycling pathway. Mol Cell Biol 24:6456–6466
Bogan JS, McKee AE and Lodish HF (2001) Insulin-responsive compartments containing Glut4 in 3T3-L1 and CHO cells: regulation by amino acid concentrations. Mol Cell Biol 21:4785–4806
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Shi, J., Kandror, K.V. (2008). Study of Glucose Uptake in Adipose Cells. In: Yang, K. (eds) Adipose Tissue Protocols. Methods in Molecular Biology™, vol 456. Humana Press. https://doi.org/10.1007/978-1-59745-245-8_23
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DOI: https://doi.org/10.1007/978-1-59745-245-8_23
Publisher Name: Humana Press
Print ISBN: 978-1-58829-916-1
Online ISBN: 978-1-59745-245-8
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