Previous work by our group has demonstrated that mesencephalic neural crest cells at an early stage of migration are able to synthesize acetylcholine (ACh). Acetylcholinesterase (AChE), the enzyme responsible for ACh degradation, was examined in neural crest cells of the chick embryo, using cytochemical and biochemical methods. Observations at the light microscope level showed that cholinesterase activity, identified as true AChE, was present at all axial levels in presumptive crest cells of the neural folds, soon after closure of the neural tube. Subsequently, AChE activity was found in cells of the individualized neural crest and in crest cells migrating at cephalic and trunk levels. Cell counts revealed that 88-94% of the total crest population was AChE-positive. Electron microscope observations indicated that the enzyme was confined to perinuclear and endoplasmic reticulum cisternae. The AChE of migrating mesencephalic neural crest cells was identified as the dimeric form (sedimentation coefficient 6.9 S) of the catalytic subunit. These results indicate that the specific AChE is present in the majority of neural crest cells all along the neural axis. Thus the ability to synthesize and degrade ACh is expressed at least in some neural crest cells at an early stage of development.