Although the involvement of nitric oxide (NO) in mediating pain and neurovascular coupling is well established, the precise mechanisms sustaining these effects are still unclear. Cyclic GMP (cGMP) probably represents the main effector of the biological effects of NO at the vascular and neuronal levels. Nitroglycerin is a NO donor, which easily crosses the blood brain barrier. Several reports have suggested that the study of nitroglycerin effects upon neuronal and cerebrovascular elements is a useful animal model for investigating the pathophysiological mechanisms underlying migraine. In this study, the anatomic distribution of cGMP in the rat brain was evaluated at serial time-points after systemic administration of nitroglycerin or vehicle. The results show an increase in cGMP immunoreactivity in the nucleus trigeminalis caudalis and in the superficial cortical arterioles 2, 3 and 4h after the drug administration. The data obtained sustains the idea that cGMP is an important mediator of nitroglycerin effect in vascular and neuronal structures that are critical elements for the transmission of cephalic pain.