Stimulating actions of imidazolines on the adrenal medulla were demonstrated by several laboratories. As only a few data about signal transduction exist, the aim of the present study is to establish a cellular model on which both subtypes of imidazoline receptors are present. Binding studies using [3H]clonidine for I1-sites and [3H]idazoxan for I2-sites were performed on bovine chromaffin cells and a PC12 cell line. The intracellular calcium signal was determined by a Fura-2 signal using a fluorescence microscope. Both subtypes of imidazoline binding sites are present on either crude membrane fractions, purified plasma membranes, and mitochondrial membranes of the adrenal medulla. Although the density of I1- and I2-sites on the plasma membrane fraction is almost equal, on the mitochondrial membrane fraction Bmax of I2-binding sites was double that of the I1-binding number. An increase in intracellular calcium signal could be obtained during stimulation of chromaffin cells by various I1- and I2-receptor modulators. Because a saturation of I2-binding could not be obtained in PC12 cells, adrenal medullary chromaffin cells may be a more suitable model for investigating imidazoline receptor signal transduction.