The topography and interrelationship of striatofugal neurons have been examined using a double retrograde tracing paradigm to label striatopallidal and striatonigral neurons in the same neostriatum. The rostral globus pallidus and the rostral substantia nigra in the same hemisphere were injected simultaneously with fluorescent tracers in three monkeys. In addition, the caudal globus pallidus and the caudal substantia nigra were injected separately in a fourth and fifth monkey with a fluorescent dye and wheat germ agglutinin-horseradish peroxidase (WGA-HRP), respectively. Digitized plots of fluorescent dye-labeled neurons revealed that large numbers of striatonigral projection neurons lie within both neostriatal nuclei, i.e., the caudate and putamen. Similarly, neurons innervating the globus pallidus were found in both caudate and putamen. The distribution of retrogradely labeled neurons observed was consistent with the topography of striatofugal projections that has been described previously, i.e., the rostrocaudal and mediolateral axes of the neostriatum are preserved in the striatopallidal and striatonigral projections (e.g., Szabo, '62, '67, '70, '72) and the dorsoventral axis is inverted in the projection of the neostriatum onto the nigra but not in the striatopallidal projection (Nauta and Domesick, '79; Gerfen, '85). Analysis of cases in which striatonigral and striatopallidal neurons were present in large numbers within the same region of the neostriatum disclosed that the two populations are intermingled such that small clusters of striatopallidal neurons are surrounded by striatonigral neurons and vice versa. The clustered arrangement of striatofugal neurons observed in the fluorescent cases was unambiguous in a case in which HRP was injected into the caudal substantia nigra. In this case, both anterogradely labeled terminals and retrogradely labeled neurons exhibited a striking, compartmental-like distribution in the posterior putamen. Our observations indicate that the matrix compartment of the neostriatum is comprised of a patchwork of interposed clusters of nigral and pallidal efferent neurons. We hypothesize that these clusters of efferent neurons may direct interdigitated cortical inputs into distinct nigro- and pallido-thalamic pathways. In view of the parallel nature of processing throughout the basal ganglia, it appears that convergence of these segregated nigral and pallidal loops must occur at the cortical level where prefrontal and premotor targets of the basal ganglia are interconnected via corticocortical projections (Selemon and Goldman-Rakic, '88).