The pattern of acetylcholinesterase (AChE) reactivity was studied in the pulvinar and dorsal lateral geniculate nucleus (DLG) of the adult macaque monkey. Discrete islands of AChE reactivity were found that correlated precisely in location with the pattern of projections from the superior colliculus and pretectum. Specifically, AChE overlies terminal fields of superior colliculus projections in the DLG, in four foci in the medial and lateral pulvinar, and in several foci in the inferior pulvinar. All of these tectal projection areas have very high AChE reactivity such that they are easily distinguished. In addition, the pretectum projects to a specific focus in the lateral pulvinar that also has a very dense AChE histochemical reaction. A number of these AChE foci could be further distinguished from other areas in the pulvinar by myeloarchitectonic characteristics. Some of the foci in the lateral and inferior pulvinars could also be distinguished by unique cytoarchitectonic features (as seen with both Nissl and Golgi stains). In an attempt to determine the possible origin of a cholinergic input to the pulvinar, horseradish peroxidase (HRP) injections and choline acetyltransferase immunohistochemistry were also done. The results of this experiment indicate that the AChE reactivity seen in the midbrain projection zones to the thalamus may be due to the precise overlap of terminal projections from the brainstem cholinergic cell groups, Ch5, Ch6, and Ch8. These results, taken together, imply that there are several anatomically and histochemically distinct zones related to extrageniculate pathways located within classically defined thalamic boundaries.