• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Nov 1, 1997; 327(Pt 3): 637–642.
PMCID: PMC1218838

GLUT4 vesicle dynamics in living 3T3 L1 adipocytes visualized with green-fluorescent protein.


Insulin stimulates glucose uptake into its target cells by a process which involves the translocation of the GLUT4 isoform of glucose transporter from an intracellular vesicular compartment(s) to the plasma membrane. The step(s) at which insulin acts in the vesicle trafficking pathway (e.g. vesicle movement or fusion with the plasma membrane) is not known. We expressed a green-fluorescent protein-GLUT4 (GFP-GLUT4) chimaera in 3T3 L1 adipocytes. The chimaera was expressed in vesicles located throughout the cytoplasm and also close to the plasma membrane. Insulin promoted a substantial translocation of GFP-GLUT4 to the plasma membrane. Time-lapse confocal microscopy demonstrated that the majority of GFP-GLUT4-containing vesicles in the basal state were relatively static, as if tethered (or attached) to an intracellular structure. A proportion (approx. 5%) of the vesicles spontaneously lost their tether, and were observed to move rapidly within the cell. Other vesicles appear to be tethered only on one edge and were observed in a rapid stretching motion. The data support a model in which GLUT4-containing vesicles are tightly tethered to an intracellular structure(s), and indicate that a primary site of insulin action must be to release these vesicles, allowing them to then translocate to and fuse with the plasma membrane.

Full Text

The Full Text of this article is available as a PDF (459K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Holman GD, Cushman SW. Subcellular localization and trafficking of the GLUT4 glucose transporter isoform in insulin-responsive cells. Bioessays. 1994 Oct;16(10):753–759. [PubMed]
  • Gould GW, Holman GD. The glucose transporter family: structure, function and tissue-specific expression. Biochem J. 1993 Oct 15;295(Pt 2):329–341. [PMC free article] [PubMed]
  • Slot JW, Geuze HJ, Gigengack S, Lienhard GE, James DE. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J Cell Biol. 1991 Apr;113(1):123–135. [PMC free article] [PubMed]
  • Slot JW, Garruti G, Martin S, Oorschot V, Posthuma G, Kraegen EW, Laybutt R, Thibault G, James DE. Glucose transporter (GLUT-4) is targeted to secretory granules in rat atrial cardiomyocytes. J Cell Biol. 1997 Jun 16;137(6):1243–1254. [PMC free article] [PubMed]
  • Martin S, Tellam J, Livingstone C, Slot JW, Gould GW, James DE. The glucose transporter (GLUT-4) and vesicle-associated membrane protein-2 (VAMP-2) are segregated from recycling endosomes in insulin-sensitive cells. J Cell Biol. 1996 Aug;134(3):625–635. [PMC free article] [PubMed]
  • Livingstone C, James DE, Rice JE, Hanpeter D, Gould GW. Compartment ablation analysis of the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 adipocytes. Biochem J. 1996 Apr 15;315(Pt 2):487–495. [PMC free article] [PubMed]
  • Clarke JF, Young PW, Yonezawa K, Kasuga M, Holman GD. Inhibition of the translocation of GLUT1 and GLUT4 in 3T3-L1 cells by the phosphatidylinositol 3-kinase inhibitor, wortmannin. Biochem J. 1994 Jun 15;300(Pt 3):631–635. [PMC free article] [PubMed]
  • Martin SS, Haruta T, Morris AJ, Klippel A, Williams LT, Olefsky JM. Activated phosphatidylinositol 3-kinase is sufficient to mediate actin rearrangement and GLUT4 translocation in 3T3-L1 adipocytes. J Biol Chem. 1996 Jul 26;271(30):17605–17608. [PubMed]
  • Tanti JF, Grémeaux T, Grillo S, Calleja V, Klippel A, Williams LT, Van Obberghen E, Le Marchand-Brustel Y. Overexpression of a constitutively active form of phosphatidylinositol 3-kinase is sufficient to promote Glut 4 translocation in adipocytes. J Biol Chem. 1996 Oct 11;271(41):25227–25232. [PubMed]
  • Tanti JF, Grillo S, Grémeaux T, Coffer PJ, Van Obberghen E, Le Marchand-Brustel Y. Potential role of protein kinase B in glucose transporter 4 translocation in adipocytes. Endocrinology. 1997 May;138(5):2005–2010. [PubMed]
  • Kohn AD, Summers SA, Birnbaum MJ, Roth RA. Expression of a constitutively active Akt Ser/Thr kinase in 3T3-L1 adipocytes stimulates glucose uptake and glucose transporter 4 translocation. J Biol Chem. 1996 Dec 6;271(49):31372–31378. [PubMed]
  • Cheatham B, Volchuk A, Kahn CR, Wang L, Rhodes CJ, Klip A. Insulin-stimulated translocation of GLUT4 glucose transporters requires SNARE-complex proteins. Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15169–15173. [PMC free article] [PubMed]
  • Volchuk A, Wang Q, Ewart HS, Liu Z, He L, Bennett MK, Klip A. Syntaxin 4 in 3T3-L1 adipocytes: regulation by insulin and participation in insulin-dependent glucose transport. Mol Biol Cell. 1996 Jul;7(7):1075–1082. [PMC free article] [PubMed]
  • Tamori Y, Hashiramoto M, Araki S, Kamata Y, Takahashi M, Kozaki S, Kasuga M. Cleavage of vesicle-associated membrane protein (VAMP)-2 and cellubrevin on GLUT4-containing vesicles inhibits the translocation of GLUT4 in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 1996 Mar 27;220(3):740–745. [PubMed]
  • Olson AL, Knight JB, Pessin JE. Syntaxin 4, VAMP2, and/or VAMP3/cellubrevin are functional target membrane and vesicle SNAP receptors for insulin-stimulated GLUT4 translocation in adipocytes. Mol Cell Biol. 1997 May;17(5):2425–2435. [PMC free article] [PubMed]
  • Dobson SP, Livingstone C, Gould GW, Tavaré JM. Dynamics of insulin-stimulated translocation of GLUT4 in single living cells visualised using green fluorescent protein. FEBS Lett. 1996 Sep 16;393(2-3):179–184. [PubMed]
  • Siemering KR, Golbik R, Sever R, Haseloff J. Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol. 1996 Dec 1;6(12):1653–1663. [PubMed]
  • Calderhead DM, Kitagawa K, Tanner LI, Holman GD, Lienhard GE. Insulin regulation of the two glucose transporters in 3T3-L1 adipocytes. J Biol Chem. 1990 Aug 15;265(23):13801–13808. [PubMed]
  • Satoh S, Nishimura H, Clark AE, Kozka IJ, Vannucci SJ, Simpson IA, Quon MJ, Cushman SW, Holman GD. 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. 1993 Aug 25;268(24):17820–17829. [PubMed]
  • Lang T, Wacker I, Steyer J, Kaether C, Wunderlich I, Soldati T, Gerdes HH, Almers W. Ca2+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy. Neuron. 1997 Jun;18(6):857–863. [PubMed]
  • Rapp S, Saffrich R, Anton M, Jäkle U, Ansorge W, Gorgas K, Just WW. Microtubule-based peroxisome movement. J Cell Sci. 1996 Apr;109(Pt 4):837–849. [PubMed]
  • Wiemer EA, Wenzel T, Deerinck TJ, Ellisman MH, Subramani S. Visualization of the peroxisomal compartment in living mammalian cells: dynamic behavior and association with microtubules. J Cell Biol. 1997 Jan 13;136(1):71–80. [PMC free article] [PubMed]
  • Navé BT, Haigh RJ, Hayward AC, Siddle K, Shepherd PR. Compartment-specific regulation of phosphoinositide 3-kinase by platelet-derived growth factor and insulin in 3T3-L1 adipocytes. Biochem J. 1996 Aug 15;318(Pt 1):55–60. [PMC free article] [PubMed]
  • Heller-Harrison RA, Morin M, Guilherme A, Czech MP. Insulin-mediated targeting of phosphatidylinositol 3-kinase to GLUT4-containing vesicles. J Biol Chem. 1996 Apr 26;271(17):10200–10204. [PubMed]
  • Ricort JM, Tanti JF, Van Obberghen E, Le Marchand-Brustel Y. Different effects of insulin and platelet-derived growth factor on phosphatidylinositol 3-kinase at the subcellular level in 3T3-L1 adipocytes. A possible explanation for their specific effects on glucose transport. Eur J Biochem. 1996 Jul 1;239(1):17–22. [PubMed]
  • Isakoff SJ, Taha C, Rose E, Marcusohn J, Klip A, Skolnik EY. The inability of phosphatidylinositol 3-kinase activation to stimulate GLUT4 translocation indicates additional signaling pathways are required for insulin-stimulated glucose uptake. Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10247–10251. [PMC free article] [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...