A line of dopamine-deficient (DD) mice was generated to allow selective restoration of normal dopamine signaling to specific brain regions. These DD floxed stop (DDfs) mice have a nonfunctional Tyrosine hydroxylase (Th) gene because of insertion of a Neo R gene flanked by lox P sites targeted to the first intron of the Th gene. DDfs mice have trace brain dopamine content, severe hypoactivity, and aphagia, and they die without intervention. However, they can be maintained by daily treatment with L-3,4-dihydroxyphenylalanine (L-dopa). Injection of a canine adenovirus (CAV-2) engineered to express Cre recombinase into the central caudate putamen restores normal Th gene expression to the midbrain dopamine neurons that project there because CAV-2 efficiently transduces axon terminals and is retrogradely transported to neuronal cell bodies. Bilateral injection of Cre recombinase into the central caudate putamen restores feeding and normalizes locomotion in DDfs mice. Analysis of feeding behavior by using lickometer cages revealed that virally rescued DDfs mice are hyperphagic and have modified meal structures compared with control mice. The virally rescued DDfs mice are also hyperactive at night, have reduced motor coordination, and are thigmotactic compared with controls. These results highlight the critical role for dopamine signaling in the dorsal striatum for most dopamine-dependent behaviors but suggest that dopamine signaling in other brain regions is important to fine-tune these behaviors. This approach offers numerous advantages compared with previous models aimed at examining dopamine signaling in discrete dopaminergic circuits.canine adenovirus ͉ feeding behavior ͉ locomotor behavior ͉ viral gene transfer ͉ tyrosine hydroxylase T he dopamine neurons in the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) comprise about two-thirds of the dopaminergic neurons in the CNS (1, 2). These midbrain dopamine neurons send dense projections to forebrain structures, including the caudate putamen (CPu), nucleus accumbens (NAc), and olfactory tubercle, as well as sparser projections to the prefrontal cortex, amygdala, and hippocampus (2). These dopaminergic circuits have been implicated in a variety of fundamental mammalian behaviors such as movement, feeding, reward responses, and learning (for review, see refs. 3-6).Dopamine-deficient (DD) mice (7) have been a useful genetic model to study the role of dopamine in these behaviors. DD mice are indistinguishable from littermates until Ϸ10 days, when they begin to manifest reduced body weight (BW) compared with controls, and, without intervention, DD mice will starve at Ϸ3 weeks (7). However, daily L-3,4-dihydroxyphenylalanine (Ldopa) treatment restores dopamine to the brain and induces Ϸ8 h of hyperactivity during which the DD mice consume enough food to survive (7,8). This ability to temporarily restore dopamine signaling with L-dopa is the primary advantage of the DD mouse model compared with the 6-hydroxydopamine-lesion model because it allows us to...