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Insulin-regulated membrane aminopeptidase (IRAP) translocates to the plasma membrane with glucose transporter-4 (GLUT4) on insulin stimulation. However, this may be impaired in patients at risk of diabetes. Recently a novel technique has been developed to assess cell surface IRAP activity dynamically using a fluorogenic membrane impermeable substrate. In this study we measured the cell surface IRAP activity and 3-O-[methyl-14C]-D-Glucose uptake in adipocytes isolated from Otsuka Long Evans Tokushima Fatty rats (OLETF), developed as a model of type 2 diabetes mellitus, to evaluate whether the translocation of GLUT4/IRAP vesicles is affected. On the addition of insulin, the cell surface IRAP activity promptly increased to reach equilibrium in a hormone dose-dependent manner. OLETF rats showed significantly lower equilibrium activity than control rats (P<0.01). Time to reach the equilibrium was also significantly longer in the OLETF case, and adipocytes isolated from OLETF rats demonstrated both a delay and a reduction in 3-O-[methyl-14C]-D-Glucose uptake. This impairment in all parameters was alleviated by treatment with pioglitazone. Continuous measurement of cell surface IRAP activity allowed accurate evaluations of GLUT4/IRAP vesicle translocation and of the establishment of defects in OLETF rats.

Impaired insulin-regulated membrane aminopeptidase translocation to the plasma membrane in adipocytes of Otsuka Long Evans Tokushima Fatty rats.