We studied the effects of prostaglandin E2 (PGE2) on hyaluronan synthesis in rabbit pericardial mesothelial cells, and the following results were obtained. (1) PGE2 (10-1000 ng/ml) stimulated hyaluronan synthesis and the level of hyaluronan synthase activity in a dose- and time-dependent manner, but PGF2 alpha did not. (2) Cyclic AMP (cAMP) levels in the cells peaked (about a 7-fold increase) at 5-10 min after adding PGE2 (1000 ng/ml). (3) Increased hyaluronan synthesis induced by PGE2 was significantly inhibited after pretreatment with either an adenylate cyclase inhibitor (2',5'-dideoxyadenosine) or a cAMP-dependent protein kinase inhibitor (PKI 5-24), but there was no inhibition with the protein kinase C inhibitor H-7. (4) When the intracellular cAMP level was raised by manipulating the levels of dibutyryl cyclic AMP or forskolin, hyaluronan synthesis and the level of hyaluronan synthase activity were also stimulated. These results suggest that PGE2 produced by cells stimulates hyaluronan synthesis in rabbit pericardial cells and that the stimulation mechanism involves the cAMP-mediated protein kinase signal transduction process.