The effects of 5-hydroxytryptamine (5-HT) and ligands selective for particular 5-HT receptor subtypes on the transmission of nociceptive information in the spinal cord are complex. In these studies, we have focused on their interactions with two endogenous mediators of pain suppression, noradrenaline (NA) and adenosine. Spinal antinociception by 5-HT is blocked by alpha-adrenoreceptor antagonists and depletion of endogenous NA by 6-hydroxydopamine, while it is potentiated by blockade of NA reuptake with desipramine. These observations provide evidence for a 5-HT receptor-mediated increase in the release of NA from the spinal cord. This action appears to be due to activation of a 5-HT1-like receptor as it is mimicked by some 5-HT1 receptor ligands (mCPP, TFMPP and 5-Me-O-DMT), but not by DOI (5-HT2) or 2-Me-5-HT (5-HT3). An additional component of 5-HT action is via release of adenosine. Antinociception by 5-HT is blocked by the adenosine receptor antagonist 8-phenyltheophylline, and 5-HT has been shown to release adenosine from the spinal cord in in vitro and in vivo paradigms. Methylxanthine-sensitive antinociception is seen with some 5-HT1 receptor ligands (CGS 12066B, mCPP), but not with others or with DOI or 2-Me-5-HT. Further characterization of the 5-HT receptor subtype involved in adenosine release will require the use of additional approaches.