Abstract
In this chapter we detail methods for the systematic dissection of GLUT4 trafficking. The methods described have been optimized for cultured 3T3-L1 adipocytes, but can be readily adapted to other cell types.
References
Burchfield JG, Lopez JA, Mele K, Vallotton P, Hughes WE (2010) Exocytotic vesicle behaviour assessed by total internal reflection fluorescence microscopy. Traffic 11(4):429–439. https://doi.org/10.1111/j.1600-0854.2010.01039.x
Burchfield JG, Lu J, Fazakerley DJ, Tan SX, Ng Y, Mele K, Buckley MJ, Han W, Hughes WE, James DE (2013) Novel systems for dynamically assessing insulin action in live cells reveals heterogeneity in the insulin response. Traffic 14(3):259–273. https://doi.org/10.1111/tra.12035
Martin S, Millar CA, Lyttle CT, Meerloo T, Marsh BJ, Gould GW, James DE (2000) Effects of insulin on intracellular GLUT4 vesicles in adipocytes: evidence for a secretory mode of regulation. J Cell Sci 113(Pt 19):3427–3438
Stenkula KG, Lizunov VA, Cushman SW, Zimmerberg J (2010) Insulin controls the spatial distribution of GLUT4 on the cell surface through regulation of its postfusion dispersal. Cell Metab 12(3):250–259. https://doi.org/10.1016/j.cmet.2010.08.005
Muretta JM, Romenskaia I, Mastick CC (2008) Insulin releases Glut4 from static storage compartments into cycling endosomes and increases the rate constant for Glut4 exocytosis. J Biol Chem 283(1):311–323. https://doi.org/10.1074/jbc.M705756200
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(14):6456–6466. https://doi.org/10.1128/MCB.24.14.6456-6466.2004
James DE, Brown R, Navarro J, Pilch PF (1988) Insulin-regulatable tissues express a unique insulin-sensitive glucose transport protein. Nature 333(6169):183–185. https://doi.org/10.1038/333183a0
Martin OJ, Lee A, McGraw TE (2006) GLUT4 distribution between the plasma membrane and the intracellular compartments is maintained by an insulin-modulated bipartite dynamic mechanism. J Biol Chem 281(1):484–490. https://doi.org/10.1074/jbc.M505944200
Fazakerley DJ, Holman GD, Marley A, James DE, Stockli J, Coster AC (2010) Kinetic evidence for unique regulation of GLUT4 trafficking by insulin and AMP-activated protein kinase activators in L6 myotubes. J Biol Chem 285(3):1653–1660. https://doi.org/10.1074/jbc.M109.051185
Wijesekara N, Tung A, Thong F, Klip A (2006) Muscle cell depolarization induces a gain in surface GLUT4 via reduced endocytosis independently of AMPK. Am J Physiol Endocrinol Metab 290(6):E1276–E1286. https://doi.org/10.1152/ajpendo.00573.2005
Brewer PD, Habtemichael EN, Romenskaia I, Mastick CC, Coster AC (2014) Insulin-regulated Glut4 translocation: membrane protein trafficking with six distinctive steps. J Biol Chem 289(25):17280–17298. https://doi.org/10.1074/jbc.M114.555714
Davey JR, Humphrey SJ, Junutula JR, Mishra AK, Lambright DG, James DE, Stockli J (2012) TBC1D13 is a RAB35 specific GAP that plays an important role in GLUT4 trafficking in adipocytes. Traffic 13(10):1429–1441. https://doi.org/10.1111/j.1600-0854.2012.01397.x
Coster AC, Govers R, James DE (2004) Insulin stimulates the entry of GLUT4 into the endosomal recycling pathway by a quantal mechanism. Traffic 5(10):763–771. https://doi.org/10.1111/j.1600-0854.2004.00218.x
Jhun BH, Rampal AL, Liu H, Lachaal M, Jung CY (1992) Effects of insulin on steady state kinetics of GLUT4 subcellular distribution in rat adipocytes. Evidence of constitutive GLUT4 recycling. J Biol Chem 267(25):17710–17715
Satoh S, Nishimura H, Clark AE, Kozka IJ, Vannucci SJ, Simpson IA, Quon MJ, Cushman SW, Holman GD (1993) Use of bismannose photolabel to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action. J Biol Chem 268(24):17820–17829
Yang J, Holman GD (1993) Comparison of GLUT4 and GLUT1 subcellular trafficking in basal and insulin-stimulated 3T3-L1 cells. J Biol Chem 268(7):4600–4603
Burchfield JG, Lopez JA, Hughes WE (2012) Using total internal reflection fluorescence microscopy (TIRFM) to visualise insulin action. In: Badoer E (ed) T visualization techniques, Neuromethods, vol 70. Humana Press, New York, pp 97–109
Schaffer DV, Koerber JT, Lim KI (2008) Molecular engineering of viral gene delivery vehicles. Annu Rev Biomed Eng 10:169–194. https://doi.org/10.1146/annurev.bioeng.10.061807.160514
Sinn PL, Sauter SL, PB MC Jr (2005) Gene therapy progress and prospects: development of improved lentiviral and retroviral vectors--design, biosafety, and production. Gene Ther 12(14):1089–1098. https://doi.org/10.1038/sj.gt.3302570
Chen C, Smye SW, Robinson MP, Evans JA (2006) Membrane electroporation theories: a review. Med Biol Eng Comput 44(1–2):5–14. https://doi.org/10.1007/s11517-005-0020-2
Sukharev SI, Klenchin VA, Serov SM, Chernomordik LV, Chizmadzhev Yu A (1992) Electroporation and electrophoretic DNA transfer into cells. The effect of DNA interaction with electropores. Biophys J 63(5):1320–1327. https://doi.org/10.1016/S0006-3495(92)81709-5
van den Hoff MJ, Moorman AF, Lamers WH (1992) Electroporation in ‘intracellular’ buffer increases cell survival. Nucleic Acids Res 20(11):2902
McKeel DW, Jarett L (1970) Preparation and characterization of a plasma membrane fraction from isolated fat cells. J Cell Biol 44(2):417–432
Simpson IA, Yver DR, Hissin PJ, Wardzala LJ, Karnieli E, Salans LB, Cushman SW (1983) Insulin-stimulated translocation of glucose transporters in the isolated rat adipose cells: characterization of subcellular fractions. Biochim Biophys Acta 763(4):393–407
Nikfarjam L, Farzaneh P (2012) Prevention and detection of mycoplasma contamination in cell culture. Cell J 13(4):203–212
Olarerin-George AO, Hogenesch JB (2015) Assessing the prevalence of mycoplasma contamination in cell culture via a survey of NCBI’s RNA-seq archive. Nucleic Acids Res 43(5):2535–2542. https://doi.org/10.1093/nar/gkv136
MacLeod RA, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG (1999) Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int J Cancer 83(4):555–563
Masters JR (2002) HeLa cells 50 years on: the good, the bad and the ugly. Nat Rev Cancer 2(4):315–319. https://doi.org/10.1038/nrc775
Shewan AM, Marsh BJ, Melvin DR, Martin S, Gould GW, James DE (2000) The cytosolic C-terminus of the glucose transporter GLUT4 contains an acidic cluster endosomal targeting motif distal to the dileucine signal. Biochem J 350(Pt 1):99–107
Shewan AM, van Dam EM, Martin S, Luen TB, Hong W, Bryant NJ, James DE (2003) GLUT4 recycles via a trans-Golgi network (TGN) subdomain enriched in Syntaxins 6 and 16 but not TGN38: involvement of an acidic targeting motif. Mol Biol Cell 14(3):973–986. https://doi.org/10.1091/mbc.E02-06-0315
Green H, Kehinde O (1975) An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell 5(1):19–27
Green H, Meuth M (1974) An established pre-adipose cell line and its differentiation in culture. Cell 3(2):127–133
Ozturk SS, Palsson BO (1990) Chemical decomposition of glutamine in cell culture media: effect of media type, pH, and serum concentration. Biotechnol Prog 6(2):121–128. https://doi.org/10.1021/bp00002a005
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Norris, D.M., Geddes, T.A., James, D.E., Fazakerley, D.J., Burchfield, J.G. (2018). Glucose Transport: Methods for Interrogating GLUT4 Trafficking in Adipocytes. In: Lindkvist-Petersson, K., Hansen, J. (eds) Glucose Transport. Methods in Molecular Biology, vol 1713. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7507-5_15
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7507-5_15
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7506-8
Online ISBN: 978-1-4939-7507-5
eBook Packages: Springer Protocols