A study has been made of the nonpyramidal, local circuit neurons in developing and mature macaque monkey prefrontal cortex with Golgi and immunocytochemical techniques. The area chosen for study is located between the cingulate gyrus and the ventral bank of the principal sulcus, and contains areas 9 and 46 as described by Walker (J. Comp. Neurol. 73:59-86, '40). In Golgi studies, the unique axonal features of impregnated neurons made possible the identification of thirteen separate classes of local circuit neurons. Five of these cell types, in their general characteristics, resembled classes identified in human prefrontal cortex, as well as in other cortical areas of macaque monkeys and other species. Measurements of the scale of axon arbors and dendritic fields of the Golgi-stained local circuit neurons also suggested particular spatial relationships of certain classes to the scale of intrinsic lattice connections made by the axons of pyramidal neurons in the same region. Similarities in morphology between cells described in human prefrontal cortex and neuron varieties described in this study indicate that this region of monkey prefrontal cortex may serve as a useful model for neuron populations in human prefrontal cortex. Sufficient morphological detail was present in immunocytochemical studies to suggest one or more identifying biochemical characteristics for seven of the thirteen classes of local circuit neurons. The calcium binding proteins, parvalbumin, calbindin D-28K, and calretinin, were found in chandelier and wide arbor neurons, neurogliaform cells, and double bouquet neurons, respectively. In addition, cholecystokinin immunoreactivity was present in medium arbor neurons and in narrow arbor cells connecting layers 2 and 4. Somatostatin 28(1-12) immunoreactivity was detected in beaded axon neurons in layers 5 and 6. This biochemical characterization of local circuit neurons, although incomplete, confirms the separate identity of at least some of the varieties distinguished by Golgi morphology, and allows a start to be made on studies examining changes in their functional state. The general inhibitory nature of these interneurons suggests that they are likely to play a crucial role in determining patterns of neural activation in the prefrontal cortex.