The ventral tegmental area (VTA) is involved in reward-related behaviours and the actions of psychostimulant drugs. It is influenced by afferents expressing a variety of neurotransmitters and neuromodulators; the innervation containing neurotensin is among the densest of these. Intra-VTA neurotensin activates dopaminergic neurons and plays an important role in the development of behavioural sensitization to psychostimulant drugs and possibly in schizophrenia. Using gold-coupled wheatgerm agglutinin as retrograde tracer in combination with nonisotopic in situ hybridization for neurotensin mRNA or neurotensin antibodies after colchicine treatment, the present study was undertaken to demonstrate the neurotensinergic neurons projecting to the VTA and determine whether (and in which subpopulations) neurotensin expression is regulated in VTA-projecting neurons after administrations of the psychostimulant drug methamphetamine or the antipsychotic haloperidol. This study reveals the lateral preoptico-rostral lateral hypothalamic continuum and the medial preoptic area as main sources for the neurotensin afferents of the VTA. Fewer neurotensinergic, VTA-projecting neurons are situated in the dorsal raphe, pedunculopontine and laterodorsal tegmental nuclei, lateral hypothalamic area, ventral endopiriform area, lateral septum, accumbens shell, parabrachial nucleus and different parts of the extended amygdala. The number of neurotensinergic VTA-projecting neurons increased significantly only after methamphetamine administration and exclusively in the accumbens shell. It is concluded that the widespread neurotensinergic VTA-projecting neurons, situated in areas involved in different reward-related behaviours, are well suited to convey distinct reward information to the VTA. The up-regulation of neurotensin expression selectively in VTA-projecting neurons in the accumbens shell following methamphetamine administration may be an important factor in the development of behavioural sensitization.