Experimental and clinical studies suggest that the low-affinity N-methyl-d-aspartate (NMDA) receptor open-channel blockers Mg and memantine are effective in reducing trigeminal nociceptive activation. The aim of the present study was to investigate the apparent effectiveness of these channel blockers using a model of trigeminal activation in vivo. Rats were anesthetized before electrically stimulating the dura mater adjacent the middle meningeal artery. Neurons responding to stimulation were recorded extracellularly using electrophysiological methods while l-glutamate or NMDA and Mg , memantine, or sodium controls were applied locally using microiontophoresis. Microiontophoretic application of Mg or memantine into the trigeminocervical complex inhibited mechanically and electrically-stimulated craniovascular afferent, l-glutamate, or NMDA-evoked neuronal activity at the second order trigeminal synapse of craniovascular afferents. By contrast, intravenous administration of MgSO (100 mg/kg) or memantine (10 mg/kg) did not significantly affect electrically-stimulated afferent-evoked activity within the trigeminocervical complex. The Mg and memantine concentrations achieved after systemic administration may not effectively inhibit activation of the trigeminocervical complex, perhaps providing an explanation for the relatively poor efficacy of these NMDA receptor open-channel blockers for headache treatment in clinical studies. Nevertheless, the present results suggest blocking of NMDA-receptor open channels inhibits nociceptive activation of the trigeminocervical complex. Further exploration of such channel blockers as a therapeutic strategy for primary head pain is warranted. This article is protected by copyright. All rights reserved.