The possible roles of the NPFF system in pain processing are summarized from the viewpoints of (1) biological activities of NPFF, (2) anatomical distribution of NPFF and its receptor(s) and (3) the regulation of NPFF and receptor(s) in animal models of pain. NPFF and NPFF analogues were found to have analgesic, pronociceptive and morphine modulating activities. Since the isolation of NPFF, several other RF-NH2 peptides have been identified and some of them were found to have nociceptive or morphine modulating activity. Depending on the pharmacological doses and locations of administration, NPFF may exhibit the biological activities of other structurally related RF-NH2 peptides thus complicating NPFF bioactivity studies and their interpretation. Acid sensing ion channels were found to respond to RF-NH2 peptides including NPFF, raising the possibility that interaction of NPFF and acid sensing ion channels can modulate nociceptive activity. NPFF and NPFF receptor mRNAs are highly expressed and localized in the superficial layers of the dorsal cord, the two genes are also in dorsal root ganglia though at much lower level. The spinal NPFF system is up-regulated by peripheral inflammation in the rat. Furthermore, immunohistochemically, NPFF receptor 2-protein was demonstrated to be increased in the primary afferents in the spinal cord of rats with peripheral inflammation. Regulation and localization of spinal NPFF systems, taken together with the analgesic bioactivity of intrathecally administered NPFF, strongly suggest involvement of spinal NPFF system in pain processing.