Electron microscope autoradiographs were prepared from diabetic rat pancreatic acini that had been incubated with 125I-insulin. Distribution histograms of the distance of the 125I-insulin silver grains from the nearest plasma membrane were prepared and compared with a histogram of an 125I line source. After 3 min of incubation, insulin was located predominately on the plasma membrane, but even at this early time 15% of the grains had an intracellular location. After 30 min of incubation, there was a decrease in grains on the plasma membrane and an increase (to 45%) in grains localized in the cell. At both times of incubation, a comparison of the distribution of the insulin grains overlying subcellular organelles to a theoretical random grain distribution pattern indicated that the intracellular distribution of insulin grains was nonrandom. At 3 min, there was a relative concentration of grains over the plasma membrane and vesicles with an average diameter of 100 nm. At 30 min, there was a concentration of grains over the plasma membrane, 100-nm vesicle, and Golgi. These studies suggest that 125I-insulin is internalized into pancreatic acinar cells in a time-dependent manner and then is nonrandomly distributed inside the cell.
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