Glutamate (GLU) associated with glycine, act as co-transmitter at the N-methyl-D-aspartate/glycine-B (NMDA/GLY(B)) receptor. Dorsal periaqueductal gray (dPAG) neurons express NMDA/GLY(B) receptors suggesting a GLU physiological role in mediating the responses elicited by stimulation of this area. Immunohistochemical data provided evidence of a possible correlation among elevated plus-maze (EPM), fear-like defensive behavior, and dPAG activity. The present data show that whereas the NMDA/GLY(B) receptor agonists increased the open-arm avoidance responses in the EPM, the antagonists had the opposite effects. Microinjection of NMDA/GLY(B) receptor agonists within the dPAG during test sessions in the EPM resulted in an enduring learned fear response detected in the retest. Therefore, in addition to the proposed role for the dPAG in panic attacks (escape), these findings suggest that the dPAG can also participate in more subtle anxiety-like behaviors.