The importance of glial cells in the generation and maintenance of neuropathic pain is becoming widely accepted. We examined the role of glial-specific gap junctions in nociception in the rat trigeminal ganglion in nerve injured and uninjured states. The connexin 43 (Cx43) gap junction subunit was found to be confined to the satellite glial cells (SGC) that tightly envelop primary sensory neurons in the trigeminal ganglion and we therefore used Cx43 RNA interference (RNAi) to alter gap junction function in SGCs. Using behavioral evaluation, together with immunocytochemical and Western blot monitoring, we show that Cx43 increased in the trigeminal ganglion in rats with a chronic constriction injury (CCI) of the infraorbital nerve. Reducing Cx43 expression using RNAi in CCI rats reduced pain-like behavior whereas in non CCI rats, reducing Cx43 expression increased pain-like behavior. The degree of pain-like behavior in CCI rats and intact, Cx43-silenced, rats was similar but systemic morphine produced analgesia only in the Cx43 RNAi treated rats. Our results support previous suggestions that increases in glial gap junctions after nerve injury increases nociceptive behavior but paradoxically the reduction of gap junctions in normal ganglia also increases nociceptive behavior possibly a reflection of the multiple functions performed by glia.