Heterogeneous distribution of glycinergic and GABAergic afferents on an identified central neuron

Immunocytochemical methods were used on serial sections to study the glycine- and gamma-amino butyric acid (GABA)ergic innervations of the teleost Mauthner (M) cell. We found different distributions for the boutons containing the two amino acids. Endings filled with GABA predominate on the distal portion of the lateral dendrite (LD) while glycine-positive profiles are more abundant on the soma and within the axon cap (AC), a specialized neuropil surrounding the M-cell initial segment. A few endings containing both transmitters are present on the soma and on the small dendrites issuing ventrally from it. At this level some glutamic acid decarboxylase (GAD)-containing boutons face glycine receptor-93 kD-associated protein, an observation suggesting that the associated glycine functions as a neurotransmitter. Elsewhere on the M-cell, where glycine and GABA are not colocalized, GAD-positive profiles were never observed in front of postsynaptic differentiations with 93 kD labelling. GABA was detected in the small vesicle boutons (SVBs), most of them, following the classification of Tuttle et al., J. Comp. Neurol. 265:254-274, 1987, belonging to the A-type, while glycine was found in the unmyelinated club endings in the AC, and in C- and B-type SVBs, outside this region. All terminals established symmetrical synapses and were filled with a population of pleiomorphic vesicles. Boutons with GABA also contained numerous dense-core vesicles suggesting the presence of an associated peptide(s). A quantitative study of the transmitter content based on the number of the gold particles revealed a variable intensity of the labelling over certain profiles. For GABA, it was maximum at the tip of the LD and it decreased proximally. In contrast, the staining density was constant for glycine along all parts of the cell, except for the ventral dendrite (VD) where it decreased progressively. Taken together, these data suggest that the amino acid content varies, depending upon the location of the synapses on their target neuron.