Glucose transporter 4 (GLUT4) is efficiently retained intracellularly. Here, we investigated the insulin-induced reduction of retention. While increasing insulin concentrations led to gradual increases in both the amount of recycling GLUT4 molecules and cell surface GLUT4 levels, the kinetics of the increase in time was independent of insulin concentration. To determine whether there are GLUT4 subpools that have a distinct insulin sensitivity, adipocytes were consecutively stimulated twice with a low concentration of insulin while recycling GLUT4 molecules were continuously labeled. This revealed that not the same pool of GLUT4 molecules was mobilized twice and thus that upon insulin stimulation, GLUT4 is likely to be recruited at random for insertion within the plasma membrane.
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