Using the double whole-cell recording technique we measured the gap junctional conductance (gj), while simultaneously monitoring the cytoplasmic free calcium concentration ([Ca2+i]) in isolated acinar cell pairs from the pancreas of mouse. Physiological concentrations of the secretagogue cholecystokinin-octapeptide (CCK-OP) elicited [Ca2+i] oscillations and after a delay in the minute range reduced gj. The results provide the first evidence suggesting that gj regulates the phase difference (delta phi) of [Ca2+i] oscillations between neighbouring cells not in an all or none process, but with decreasing absolute value of gj delta phi monotonically increases. CCK-OP induced electrical uncoupling as well as the corresponding increase of delta phi could be blocked completely by the protein kinase C inhibitor polymyxin B. The data support the hypothesis that Ca2+ flow through gap junctions synchronizes [Ca2+i] oscillations between jap junctional coupled cells.