ACTH sensitivity of adenyl cyclase in mitochondrial and microsomal fractions from bovine adrenal cortex and in ghosts from rat fat cells can be abolished by the Ca complexing agent EGTA and restored by addition of Ca. EGTA also increases basal levels of cyclase activity in both systems and the degree of epinephrine stimulation in ghosts. Added Ca ion is inhibitory for basal and hormone stimulated activity in both systems. Ca may thus regulate 3′,5′-AMP formation both in the absence and presence of hormones. © 1969.