The Journal of Cell Biology, Vol 113, 123-135, Copyright © 1991 by The Rockefeller University Press

ARTICLES

Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat

JW Slot, HJ Geuze, S Gigengack, GE Lienhard and DE James
Department of Cell Biology, Medical School, University of Utrecht, The Netherlands.

Antibodies specific for the insulin-regulatable glucose transporter (GLUT 4) were used to immunolocalize this protein in brown adipose tissue from basal- and insulin-treated rats. Cryosections of fixed tissue were incubated with antibodies, which were subsequently labeled with Protein A/gold and examined by EM. Antibodies against albumin and cathepsin D were also used with gold particles of different sizes to identify early and late endosomes, respectively. Under basal conditions 99% of the GLUT 4 labeling was located within the cell. Labeling was predominantly in the trans-Golgi reticulum and tubulo-vesicular structures elsewhere in the cytoplasm. In insulin-stimulated cells approximately 40% of the GLUT 4 labeling was at the cell surface, where it was randomly distributed, except for occasional clustering in coated pits. Moreover, after insulin treatment, GLUT 4 was also enriched in early endosomes. We conclude that translocation of GLUT 4 to the cell surface is the major mechanism by which insulin increases glucose transport. In addition, these results suggest that in the presence of insulin GLUT 4 recycles from the cell surface, probably via the coated pit-endosome pathway that has been characterized for cell surface receptors, and also that insulin causes the redistribution of GLUT 4 by stimulating exocytosis from GLUT 4-containing tubulo-vesicular structures, rather than by slowing endocytosis of GLUT 4.
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• Clemens, D. L., Lee, B.-Y., Horwitz, M. A. (2002). The Mycobacterium tuberculosis Phagosome in Human Macrophages Is Isolated from the Host Cell Cytoplasm. Infect. Immun. 70: 5800-5807 [Abstract] [Full Text]  
• Udby, L., Calafat, J., Sorensen, O. E., Borregaard, N., Kjeldsen, L. (2002). Identification of human cysteine-rich secretory protein 3 (CRISP-3) as a matrix protein in a subset of peroxidase-negative granules of neutrophils and in the granules of eosinophils. J. Leukoc. Biol. 72: 462-469 [Abstract] [Full Text]  
• Konrad, D., Bilan, P. J., Nawaz, Z., Sweeney, G., Niu, W., Liu, Z., Antonescu, C. N., Rudich, A., Klip, A. (2002). Need for GLUT4 Activation to Reach Maximum Effect of Insulin-Mediated Glucose Uptake in Brown Adipocytes Isolated From GLUT4myc-Expressing Mice. Diabetes 51: 2719-2726 [Abstract] [Full Text]  
• Mohrmann, K., Gerez, L., Oorschot, V., Klumperman, J., van der Sluijs, P. (2002). rab4 Function in Membrane Recycling from Early Endosomes Depends on a Membrane to Cytoplasm Cycle. J. Biol. Chem. 277: 32029-32035 [Abstract] [Full Text]  
• Pearson, M. M., Lafontaine, E. R., Wagner, N. J., St. Geme III, J. W., Hansen, E. J. (2002). A hag Mutant of Moraxella catarrhalis Strain O35E Is Deficient in Hemagglutination, Autoagglutination, and Immunoglobulin D-Binding Activities. Infect. Immun. 70: 4523-4533 [Abstract] [Full Text]  
• Oorschot, V., de Wit, H., Annaert, W. G., Klumperman, J. (2002). A Novel Flat-embedding Method to Prepare Ultrathin Cryosections from Cultured Cells in Their In Situ Orientation. J. Histochem. Cytochem. 50: 1067-1080 [Abstract] [Full Text]  
• Sancho, M. C., Schleich, S., Griffiths, G., Krijnse-Locker, J. (2002). The Block in Assembly of Modified Vaccinia Virus Ankara in HeLa Cells Reveals New Insights into Vaccinia Virus Morphogenesis. J. Virol. 76: 8318-8334 [Abstract] [Full Text]  
• Bulow, E., Bengtsson, N., Calafat, J., Gullberg, U., Olsson, I. (2002). Sorting of neutrophil-specific granule protein human cathelicidin, hCAP-18, when constitutively expressed in myeloid cells. J. Leukoc. Biol. 72: 147-153 [Abstract] [Full Text]  
• Canonne-Hergaux, F., Calafat, J., Richer, E., Cellier, M., Grinstein, S., Borregaard, N., Gros, P. (2002). Expression and subcellular localization of NRAMP1 in human neutrophil granules. Blood 100: 268-275 [Abstract] [Full Text]  
• Huber, D., Cramer, E. M., Kaufmann, J. E., Meda, P., Masse, J.-M., Kruithof, E. K. O., Vischer, U. M. (2002). Tissue-type plasminogen activator (t-PA) is stored in Weibel-Palade bodies in human endothelial cells both in vitro and in vivo. Blood 99: 3637-3645 [Abstract] [Full Text]  
• Keller, S. R., Davis, A. C., Clairmont, K. B. (2002). Mice Deficient in the Insulin-regulated Membrane Aminopeptidase Show Substantial Decreases in Glucose Transporter GLUT4 Levels but Maintain Normal Glucose Homeostasis. J. Biol. Chem. 277: 17677-17686 [Abstract] [Full Text]  
• Kupriyanova, T. A., Kandror, V., Kandror, K. V. (2002). Isolation and Characterization of the Two Major Intracellular Glut4 Storage Compartments. J. Biol. Chem. 277: 9133-9138 [Abstract] [Full Text]  

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