Objective: Migraine is a unilateral, pulsating and very disabling half-sited pain to the head which is in part due to dilatation of the cranial arteries. Clinical trials show that prostaglandin E2 (PGE2) induces headache. Furthermore, during a migraine-attack, PGE2 is found in increased levels in blood and saliva of migraineurs. This study investigates the role of PGE2 receptors (EP1-4) in the large rat cranial arteries in relation to migraine. Methods: We investigated the dilatory effect of PGE2, EP2 and EP4 agonists on the preclinical "closed cranial window" in vivo model on the middle meningeal (MMA) and middle cerebral artery (MCA). Myographs were used for confirmation in vitro. Antagonists for each receptor were used to estimate the contribution of the receptors. RT-PCR and Western blotting were used to quantify and detect the receptors in the arteries. Results: Dilatation to butaprost (EP2 agonist) (Emax 110±18%, pED50 5.0±0.17) was less pronounced compared to PGE2 (Emax 207±43%, pED50 7.0±0.31) and ONO-AE1-329 (EP4 agonist) (127±15%, pED50 7.4±0.09) in middle meningeal artery (MMA) in vivo. BGC20-1531 (EP4 receptor antagonist), AH6809 (EP2 receptor antagonist) and SQ22536 (adenylate cyclase inhibitor) significantly inhibited the PGE2 induced vasodilatory response in rats. Likewise, the used antagonists significantly inhibited the PGE2 relaxation in rat MMA and MCA in vitro. Conventional RT-PCR and Western blotting showed that all PGE2 receptors (EP1-EP4) mRNA were expressed in rat neuronal tissues and cerebral arteries. However, quantification of the mRNA expression profile of the dilatory receptors (EP2 and EP4) showed dominance of these receptors in rat MMA and MCA as compared to the investigated neuronal tissues. Conclusions: PGE2 induced vasodilatory responses and could be inhibited by EP2 and EP4 receptor antagonists, possibly via cAMP mechanisms. The EP2 and EP4 receptors are predominant in the arteries. Thus, these receptors might be a putative target in the development of anti-migraine drugs.