GABA and glycine immunolabeling in the chicken tangential nucleus

In the vestibular nuclei, GABAergic and glycinergic neurons play important roles in signal processing for normal function, during development, and after peripheral vestibular lesions. The chicken tangential nucleus is a major avian vestibular nucleus, whose principal cells are projection neurons with axons transmitting signals to the oculomotor nuclei and/or cervical spinal cord. Antibodies against GABA, glycine and glutamate were applied to study immunolabeling in the tangential nucleus of 5-7 days old chicken using fluorescence detection and confocal imaging. All the principal cells and primary vestibular fibers were negative for GABA and glycine, but positive for glutamate. GABA is the predominant inhibitory neurotransmitter in the tangential nucleus, labeling most of the longitudinal fibers in transverse tissue sections and more than 50% of all synaptic terminals. A large fraction of GABAergic terminals were derived from the longitudinal fibers, with fewer horizontal GABAergic fibers detected. GABA synapses terminated mainly on dendrites in the tangential nucleus. In contrast, glycine labeling represented about one-third of all synaptic terminals, and originated from horizontally-coursing fibers. A distinct pool of glycine-positive terminals was found consistently around the principal cell bodies. While no GABA or glycine-positive neuron cell bodies were found in the tangential nucleus, several pools of immunopositive neurons were present in the neighboring vestibular nuclei, mainly in the descending vestibular and superior vestibular nuclei. GABA and glycine double-labeling experiments revealed little colocalization of these two neurotransmitters in synaptic terminals or fibers in the tangential nucleus. Our data support the concept of GABA and glycine playing critical roles as inhibitory neurotransmitters in the tangential nucleus. The two inhibitory neurotransmitters have distinct and separate origins and display contrasting subcellular termination patterns, which underscore their discrete roles in vestibular signal processing.