Background Prostaglandins (PGs), particularly prostaglandin D₂ (PGD₂), E₂ (PGE₂), and I₂ (PGI₂), are considered to play a role in migraine pain. In humans, infusion of PGD₂ causes lesser headache as compared to infusion of PGE₂ and PGI₂. Follow-up studies in rats have shown that infusion of PGE₂ and PGI₂ dilate the middle meningeal artery (MMA), and mRNA for PGE₂ and PGI₂ receptors is present in rat trigeminovascular system (TVS) and in the brain structures associated with pain. In the present study, we have characterized the dilatory effect of PGD₂ on rat MMA and studied the relative mRNA expression of PGD₂ receptors and lipocalin-type of PGD₂ synthase (L-PGDS). Method Rat closed-cranial window (CCW) model was used to study the effect of the DP₁ receptor antagonist, MK-0524, on PGD₂-induced vasodilation of middle meningeal artery. The qPCR technique was used for mRNA expression analysis. Results PGD₂ infusion evoked a dose-dependent dilation of the rat MMA. The calculated mean pED₅₀ value was 5.23 ± 0.10 and E_max was 103 ± 18% (n = 5). MK-0524 significantly (∼61%, p < 0.05) blocked the PGD₂-induced dilation of MMA. mRNA for the DP₁, DP₂ and L-PGDS were expressed differentially in all tested tissues. DP₁ receptor mRNA was expressed maximally in trigeminal ganglion (TG) and in cervical dorsal root ganglion (DRG). Conclusions High expression of DP₁ mRNA in the TG and DRG suggest that PGD₂ might play a role in migraine pathophysiology. Activation of the DP₁ receptor in MMA was mainly responsible for vasodilation induced by PGD₂ infusion.