Dissecting GLUT4 traffic components in L6 myocytes by fluorescence-based, single-cell assays.
Postprandial blood glucose homeostasis is regulated by an insulin-stimulated increase in glucose transport into muscle and fat tissues via glucose transporter isoform 4 (GLUT4). In the basal state, this constitutively recycling membrane protein predominantly resides intracellularly. In order to achieve the insulin-stimulated increase in glucose flux, GLUT4 increases its cell surface abundance at the expense of preformed intracellular depots. By confocal microscopy of cultured L6 muscle cells stably expressing myc-tagged GLUT4 (L6-GLUT4myc), we can visualize the two arms of GLUT4 traffic: exocytosis (movement to the cell surface) and endocytosis (internalization from the cell surface).
Antonescu
Costin N
CN
Hospital for Sick Children, Toronto, Ontario, Canada.
Randhawa
Varinder K
VK
Klip
Amira
A
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004705:Endocytosis / Q000187:drug effects; D005453:Fluorescence; D005455:Fluorescent Antibody Technique; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D032342:Muscle Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors
D051275:Glucose Transporter Type 4; D007328:Insulin; D011993:Recombinant Fusion Proteins