The present paper describes a unique distribution of true AChE activity in the IO. In the dorsal accessory olive three areas with high AChE activity can be distinguished. The medial accessory olive can be subdivided into a caudal part which shows rostro-caudally directed bands with different enzymatic activity, and a rostral part which shows a more uniform, medium activity. In the nucleus beta and the dorso-medial cell column, AChE activity is low. The ventral and dorsal lamellae of the principal olive contain areas with high, medium, and low activity. The dorsal cap is strongly positive, while the ventrolateral outgrowth is negative for AChE. Enzyme distribution cannot be fully explained on the base of the known afferent and efferent connections with the IO. However, histochemical results provide evidence that generally supports a subdivision of the IO that mirrors these connections (Brodal, '40; Armstrong et al., '74' Boesten and Voogd, '75; Groenewegen et al., '75).