DescriptionThe pedunculopontine nucleus (PPN) has traditionally been studied within the context of the midbrain locomotor region (MLR) (Shik et al., 1966; Grillner and Shik 1973; Mori et al., 1977 , Skinner and Garcia-Rill 1984), a descending motor control circuit. Yet, the dopaminergic nuclei of the substantia nigra and ventral tegmental area (VTA) receive the highest density of excitatory fibers from the PPN, most notably the substantia nigra pars compacta (SNc) (Dautan et al., 2021). While dopamine has implications in movement and exploratory behavior (Fink and Smith, 1980), the effect of PPN excitatory input on the SNc/VTA circuit remains poorly understood. The experiments herein address the organization of the glutamatergic (i.e. VGluT2+) PPN-to-substantia nigra efferent circuit in order to better understand the relationship between PPN and basal ganglia. Notably, excitatory circuits that originate in the PPN and terminate in specific basal ganglia nuclei appear to be segregated into separate circuits. Specifically, SNc- projecting glutamatergic PPN neurons do not project to the striatum except for rare exceptions. SNc-projecting PPN glutamatergic neurons collateralize to innervate specific nodes of the basal ganglia including the subthalamic nucleus (STN) and globus pallidus, but typically not the striatum. Anterograde transsynaptic labelling using an AAV1 construct (Zingg et al., 2017; Zingg et al., 2020) provides evidence that specific PPN neurons innervate segregated SNc/VTA subpopulations that project to unique subregions of the striatum, suggesting that the topography of the PPN may correspond to functional differences in basal ganglia control coincidentally observed along the dorsoventral axis of the striatum. Taken together, these findings suggest a model of PPN wherein heterogeneous subpopulations of VGluT2+ neurons form the basis for specific downstream effects on basal ganglia function via dopamine release and the feed- forward excitatory control of specific basal ganglia nodes.