The purinergic receptor P2X₃, expressed in the central terminals of primary nociceptive neurons in the brainstem, plays an important role in pathological pain. However, little is known about expression of P2X₃ in the brainstem astrocytes and its involvement in craniofacial pathologic pain. To address this issue, we investigated the expression of P2X₃ in astrocytes in the trigeminal caudal nucleus (Vc) in a rat model of craniofacial neuropathic pain, chronic constriction injury of infraorbital nerve (CCI-ION). We found that 1) P2X₃-immunoreactivity is observed in the brainstem astrocytes, preferentially in their fine processes, 2) the number of P2X₃-positive fine astrocytic processes and the density of P2X₃ in these processes were increased significantly in CCI-ION rats, compared to control rats, and 3) administration of MPEP, a specific mGluR5 antagonist, alleviated the mechanical allodynia and abolished the increase in density of P2X₃ in fine astrocytic processes caused by CCI-ION. These findings reveal preferential expression of P2X₃ in the fine astrocytic processes in the brainstem, propose a novel role of P2X₃ in the fine astrocytic process in the mechanism of craniofacial neuropathic pain, and suggest that the expression of astrocytic P2X₃ may be regulated by astrocytic mGluR5.