Release of Neuropeptide FF, an Anti‐Opioid Peptide, in Rat Spinal Cord Slices Is Voltage‐ and Ca2+‐Sensitive: Possible Involvement of P‐Type Ca2+ Channels

: Neuropeptide FF (NPFF), an FMRFamide‐like peptide with antiopioid properties, inhibits morphine‐induced analgesia but also produces hyperalgesia. In the present study, the mechanisms of NPFF release were investigated in an in vitro superfusion system with rat spinal cord slices. The opening of voltage‐sensitive Na+ channels with veratridine (20 µM) induced calcium‐dependent NPFF release, which was abolished by tetrodotoxin (1 µM), suggesting that NPFF release depends on nerve impulse activity. We also showed that NPFF release was a function of the extent of depolarization and was calcium dependent. The 30 mM K+‐induced release was blocked by Co2+ or Ni2+ (2.5 mM) but was unaffected by Ca2+ channel blockers of the L type—Cd2+ (100 µM), nifedipine or nimodipine (10 µM), diltiazem (20 µM), or verapamil (50 µM)—or the N type—ω‐conotoxin GVIA (1 µM). In contrast, ω‐agatoxin IVA (1 µM) led to a 65% reduction in NPFF release, suggesting that P‐type Ca2+ channels play a prominent role. The 35% remaining release resulted from activation of an unknown subtype. The NPFF‐like material in superfusates recognized spinal NPFF receptors, suggesting that NPFF release in the spinal cord has a physiological role.