The ventral tegmental area of the rat brain has been shown to possess high densities of neurotensin- and dopamine-containing neuronal perikarya. We recently demonstrated that microinjection of neurotensin into the ventral tegmental area produces behavioral hyperactivity similar to amphetamine-induced increase in exploratory behaviors, but lacking stereotypies. In this study, we report that the threshold dose for neurotensin-induced hyperactivity is 0.10-0.25 micrograms neurotensin/side. Either intracerebroventricular injection of haloperidol (5.0 micrograms/lateral ventricle) or destruction of the mesolimbic dopamine system by 6-hydroxydopamine abolishes the behavioral hyperactivity produced by intraventral tegmental injection of neurotensin (2.5 micrograms/side). Using high pressure liquid chromatography with electrochemical detection, we show that neurotensin injection into the ventral tegmental area increases the concentration of dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid in the nucleus accumbens and olfactory tubercles, but not in the striatum. This effect is especially profound in the nucleus accumbens where the threshold dose is less than 0.025 micrograms/side. The ratio of 3,4-dihydroxyphenylacetic acid to dopamine increased in the nucleus accumbens and olfactory tubercles in a dose-dependent fashion (0.025 microgram-2.50 micrograms/side). Neurotensin-induced behavioral hyperactivity correlates positively with neurotensin-induced changes in the ratio of 3,4-dihydroxyphenylacetic acid to dopamine. This study indicates that neurotensin acts in the ventral tegmental area to activate the mesolimbic dopamine system. Further, this activation produces behavioral hyperactivity characterized by an increase in exploratory behaviors. The fact that both immunoreactive neurotensin and neurotensin receptors are found in high concentration in the ventral tegmental area supports the possible physiological significance of this peptide-catecholamine interaction.