Transverse sections through various levels of the feline gastrointestinal tract (antrum pyloricum, duodenum, jejunum, ileum and colon) were obtained from animals perfused with a mixture of glutaraldehyde and paraformaldehyde, and incubated with an antiserum specific for glutaraldehyde-fixed GABA. This antiserum has previously been characterized and shown to react selectively with presumed GABAergic neurons in the CNS. At all levels of the gastrointestinal tract, staining was observed in varicose fibers in the myenteric plexus and the circular muscle layer. Staining was abolished by glutaraldehyde complexes of GABA previously added to the antiserum, but not by similar complexes of related amino acids. The findings strongly suggest that GABA is concentrated in a subpopulation of gastrointestinal nerve fibers and add to previous evidence favoring a transmitter role for GABA in the enteric nervous system.