Effect of PACAP on cAMP level in the rat cerebral microvessels was investigated. The cerebral microvessels were prepared from rat cerebral cortex by albumin flotation and glass beads filtration technique. When the microvessels were incubated with PACAP 27, PACAP 38 and VIP, cAMP in the microvessels was increased rapidly reaching a plateau value within 60 s. PACAP 27, PACAP 38 and VIP increased cAMP level in a dose-dependent manner with EC50 values of 4.7, 7.0 and 34 nM, respectively. These results suggest that PACAPs play a role in the regulation of the cerebral microvessel function.