Rapid insulin-induced exocytosis in white rat adipocytes

Insulin is believed to increase glucose permeability of adipocytes by regulating the incorporation of glucose transporters into the plasma membrane by exocytosis. This process involves fusion of membrane-bound cellular compartments with the plasma membrane, thus influencing the plasma membrane area. However, insulin-induced changes in plasma membrane area have not yet been demonstrated. In the present study we monitored fluorescence intensity with a confocal microscope to study the effect of insulin on adipocyte plasma membrane area. After cell isolation and adhesion to a glass cover-slip, adipocytes were stained with the dye FM1-43, a membrane area reporter. At rest, the rate of fluorescence intensity increase was initially high, but gradually stabilized at 2%/min. This steady increase in fluorescence is due to a slow rate of exocytosis coupled to endocytosis, since the removal of FM1-43 from the bath did not abolish FM1-43 fluorescence. Insulin addition caused an abrupt increase of fluorescence intensity of 4%/min, which was significantly higher than in controls. These results suggest rapid, insulin-induced incorporation of new membrane into the plasma membrane by exocytosis.