Intra-acinar zymogen activation is an early and critical feature of acute pancreatitis. High sustained calcium elevations within the pancreatic acinar cell precede pathologic zymogen activation. However, the determinants of the calcium signal are not clear. We have previously shown that cAMP sensitizes the pancreatic acinar cell to zymogen activation. In this study, we examined the role of cAMP in shaping the dynamics of the calcium signal. We hypothesized that cAMP regulates intra-acinar calcium channel opening in response to calcium-evoking agonists. In these experiments, isolated acinar cells were loaded with the calcium dye fluo4 and imaged using a confocal laser scanning microscope. They were stimulated with the calcium-evoking agonist carbachol (1 uM) in the absence or presence of the cAMP analog 8-Br-cAMP (100 uM). We noted that the rise time after carbachol stimulation was reduced by 55% (p < 0.005) in the basal region of the cell and to a lesser degree in the apical region. In addition, the slope of the calcium rise was increased 2.8 and 1.9 fold in the basal and apical regions, respectively, with cAMP pretreatment (p < 0.01). Furthermore, the speed of the apical-to-basolateral calcium wave increased from 8 to 21 um/sec (p < 0.05). Neither the baseline calcium wave speed nor the cAMP-mediated increase was affected by calcium-free media with EGTA (2 mM). However, dantrolene (100 uM), an inhibitor of the intra-cellular calcium channel the ryanodine receptor, abrogated the cAMP-mediated increase in wave speed (p < 0.05). No differences were noted in the peak calcium response in the apical or basal regions. These data suggest that cAMP affects the speed of the apical to basal calcium wave. Further, the effects of cAMP on the acinar cell calcium wave appear to be dependent on ryanodine receptor opening. We conclude that cAMP appears to regulate ryanodine receptor calcium opening in pancreatic acinar cells.