The interrelationship between arginine and calcium in glucagon release was studied using the in vitro perfused rat pancreas. In the presence of a normal 2 m M calcium concentration, 10 m M arginine provoked biphasic glucagon release, the amplitude of such a response being inversely related to the glucose concentration of the perfusate. At the lowest 3.3 m M glucose concentration, and after prolonged (40 min) as well as short (10 min) periods of calcium deprivation, arginine-induced glucagon release was inhibited. These results clearly indicate that calcium is necessary for either the recognition or effectiveness of arginine as a stimulus to the α-cell. Taking into account this, as well as other data presently available in the literature, it is suggested that calcium plays a versatile role – permissive, inhibitory and stimulatory – in glucagon secretion. The intimate mechanisms by which calcium exerts such versatile actions in the α-cell remain to be elucidated.