Insulin recruits GLUT4 from specialized VAMP2-carrying vesicles as well as from the dynamic endosomal/trans-Golgi network in rat adipocytes

G Ramm, J W Slot, D E James, W Stoorvogel

    Research output: Contribution to journalArticleAcademicpeer-review

    Abstract

    Insulin treatment of fat cells results in the translocation of the insulin-responsive glucose transporter type 4, GLUT4, from intracellular compartments to the plasma membrane. However, the precise nature of these intracellular GLUT4-carrying compartments is debated. To resolve the nature of these compartments, we have performed an extensive morphological analysis of GLUT4-containing compartments, using a novel immunocytochemical technique enabling high labeling efficiency and 3-D resolution of cytoplasmic rims isolated from rat epididymal adipocytes. In basal cells, GLUT4 was localized to three morphologically distinct intracellular structures: small vesicles, tubules, and vacuoles. In response to insulin the increase of GLUT4 at the cell surface was compensated by a decrease in small vesicles, whereas the amount in tubules and vacuoles was unchanged. Under basal conditions, many small GLUT4 positive vesicles also contained IRAP (88%) and the v-SNARE, VAMP2 (57%) but not markers of sorting endosomes (EEA1), late endosomes, or lysosomes (lgp120). A largely distinct population of GLUT4 vesicles (56%) contained the cation-dependent mannose 6-phosphate receptor (CD-MPR), a marker protein that shuttles between endosomes and the trans-Golgi network (TGN). In response to insulin, GLUT4 was recruited both from VAMP2 and CD-MPR positive vesicles. However, while the concentration of GLUT4 in the remaining VAMP2-positive vesicles was unchanged, the concentration of GLUT4 in CD-MPR-positive vesicles decreased. Taken together, we provide morphological evidence indicating that, in response to insulin, GLUT4 is recruited to the plasma membrane by fusion of preexisting VAMP2-carrying vesicles as well as by sorting from the dynamic endosomal-TGN system.

    Original languageEnglish
    Pages (from-to)4079-91
    Number of pages13
    JournalMolecular Biology of the Cell
    Volume11
    Issue number12
    Publication statusPublished - Dec 2000

    Keywords

    • Adipocytes
    • Animals
    • Cell Compartmentation
    • Cells, Cultured
    • Endosomes
    • Glucose Transporter Type 4
    • Insulin
    • Male
    • Membrane Proteins
    • Microscopy, Immunoelectron
    • Monosaccharide Transport Proteins
    • Muscle Proteins
    • R-SNARE Proteins
    • Rats
    • Transport Vesicles
    • trans-Golgi Network

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