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Rat pancreatic beta and alpha 2 cells were purified by autofluorescence-activated cell sorting and used for electrophysiological patch clamp studies and measurements of the initial uptake of 45Ca. Both beta and alpha 2 cells were electrically active, the action potentials of the latter cells also were detected in the absence of glucose. Furthermore, alpha 2 cells differed from beta cells in lacking a glucose-sensitive K+ channel with a single conductance of 50-60 pS (in symmetric 140 mM K+ solutions). The rate of Ca2+ entry into the alpha 2 cells was slower than that into the beta cells, being equivalent to 0.2 mmol, kg-1 dry wt min-1. Whereas raising the glucose concentration to 20 mM significantly increased the amount of Ca2+ entering the beta cells, the sugar was without effect on Ca2+ entry into the alpha 2 cells.