The activity of cytochrome oxidase (CO), a mitochondrial enzyme of oxidative energy metabolism, is regulated by nerve cell functional activity. The mechanism of CO activity regulation was studied using histochemical and immunohistochemical methods to show the distributions of CO activity and protein, respectively, in the macaque monkey visual system under normal and experimental conditions. In normal animals, patterns of CO activity were found to reflect underlying patterns of CO protein distribution; for example, puffs of high CO activity in cortical area 17 contained high levels of CO protein. Experimental animals were injected monocularly with TTX for 3–4 weeks; this treatment blocks retinal impulses in the injected eye and results in decreased CO activity in lateral geniculate laminae and striate cortical columns normally driven by the treated eye. The experimentally induced decreases in CO activity were also found to reflect underlying parallel decreases in CO protein levels. These results suggest that CO activity is regulated mainly at the level of the local amount rather than the turnover number of the enzyme and imply that the rates of CO synthesis and/or degradation are regulated by neural functional activity.