The enteric nervous system is formed from neural crest-derived cells. These cells enter the gut, migrate, proliferate, and ultimately differentiate into neurons and glia. We have used a human anti-neuronal autoantibody (ANNA-1), which recognizes neuron-specific RNA-binding proteins of the Hu family as an early marker of neuronal phenotype, to study the appearance of enteric neurons in the developing chicken gut. Immunoreactive cells appear first in the gizzard primordium at E3.5 and are found at progressively more caudal locations in the gut as development proceeds. Nascent neurons are present at the yolk stalk at E4.5, at the ileocecal junction at E6.5, and within the rectum at E7.5-8.5. Neurons appear slightly later in the esophagus. Aggregates of cells resembling developing ganglia were first observed at E6.5 in the distal esophagus and at E8.5 in the proximal esophagus. A small number of cells appeared in the vagus nerve trunks at E4.5 and that number increased at E7.5-8.5. Immunoreactive cells were also found in the sympatho-aortic plexus between the mesonephri and in the dorsal mesentery. These cells appeared to coalesce and form the ganglionated Nerve of Remak which contained positive cells at E3.5. This Nerve extended to the yolk stalk at E4.5 and to duodenum at E6.5. We conclude that the appearance of nascent neurons occurs first in the gizzard and proceeds more rapidly in a distal than proximal direction along the gut. Furthermore, cell that appear to be nascent neurons are found in the vagus and in the dorsal mesentery.