In vivo treatment with an antisense (AS) phosphorothioate oligodeoxynucleotide (oligo) to the rat kappa-opioid receptor selectively inhibited kappa-mediated analgesia in the rat cold-water tail-flick test. Intracerebroventricular (i.c.v.) AS oligo significantly inhibited the analgesic effect of i.c.v. spiradoline, but not that of mu- or delta-opioid agonists. The dose-effect curve for s.c. spiradoline was shifted to the right after AS, but not missense or sense oligo treatment. Thus, AS oligos provide another technique with which to selectively manipulate opioid receptors and further support the role of non-mu opioid receptors in mediating analgesia in rats.