Although the structure and function of primate prefrontal cortex undergo substantial modifications during postnatal development, relatively little is known about the maturation of neurotransmitter systems in these cortical regions. In the primate brain, cholecystokinin is present in the greatest concentrations in prefrontal regions. Thus, in this study, we used immunohistochemical techniques to investigate the postnatal development of the cholecystokinin innervation of monkey prefrontal cortex. In animals aged 4 days through adult, cholecystokinin immunoreactivity was present in nonpyramidal neurons that appeared to represent at least two distinct cell types. The most common type was a vertically oval bitufted neuron, located in layers II-superficial III, which typically had a radially descending axon that gave rise to short collaterals in layer IV. Another frequently observed cell type was a larger multipolar neuron located in the superficial half of layer III. The axon of these neurons branched locally in the vicinity of the cell body. The greatest density of cholecystokinin-containing neurons and processes was present in monkeys less than 1 month of age. The density of immunoreactive structures in every prefrontal region then progressively declined with increasing age, with the most marked changes occurring during the first postnatal year. As a result, the density of labeled neurons in adult monkeys was less than one-third of that in neonatal monkeys. However, labeled structures were significantly more dense in some ventromedial and orbital regions than in dorsal regions of the prefrontal cortex in neonatal, but not in older animals. In all animals, cholecystokinin-containing neurons were present in highest density in layers II-superficial III, and labeled terminal fields were observed in layers II, IV, and VI. In animals less than 1 month of age, fascicles of radial fibers traversed through layers III and V, whereas in animals 1 to 3 months of age, individual radial fibers rather than fiber bundles were present in layers III and V. In addition, immunoreactive pericellular arrays, which appeared to surround unlabeled nonpyramidal cells, were present in layers V and VI and the subcortical white matter in the youngest monkeys. Although many aspects of the cholecystokinin innervation of monkey prefrontal cortex remain constant during postnatal life, the distinct developmental changes in the cholecystokinin innervation of these regions suggest that it may play an important role in the maturation of the cortical circuitry that mediates the acquisition of certain cognitive abilities.