In this study, we examined the antinociceptive effect of beta-lactotensin, a neurotensin agonist that has been isolated from the chymotrypsin digest of beta-lactoglobulin as an ileum-contracting peptide. Beta-lactotensin showed naloxone-insensitive antinociceptive activity by the tail-pinch test after i.c.v. (200 nmol/mouse) or s.c. (300 mg/kg) administration in ddY mice. Tolerance was not developed to antinociception induced by beta-lactotensin after repeated s.c. administration for 5 days. The antinociceptive activity of beta-lactotensin was blocked by treatment with the neurotensin NT2 receptor antisense ODN, while treatment with the NT1 receptor antisense ODN had no effect. The antinociceptive activity was also blocked by a dopamine D1 receptor antagonist, SCH23390 (1 microg/mouse, i.c.v.), while a D2 receptor antagonist, raclopride (0.5 microg/mouse, i.c.v.), did not block the activity. These results indicate that the antinociceptive activity of beta-lactotensin is mediated by NT2 and D1 receptors.