The organization of glycine-like immunoreactive (GLY-LIR) processes was investigated within the sympathetic preganglionic neuropils of male Sprague-Dawley rats and pigeons (Columba livia). Sympathetic preganglionic neurons were retrogradely labeled with horseradish peroxidase following injections into the superior cervical ganglion in rats or into the avian homologue of the mammalian stellate ganglion (paravertebral ganglion 14) in pigeons. Glycine-like immunoreactivity was visualized using postembedding immunoperoxidase and immunogold labeling methods. The neuropils surrounding pigeon sympathetic preganglionic neurons in the principal preganglionic cell column (nucleus of Terni) and in the nucleus intercalatus contained numerous GLY-LIR puncta. Many of these processes appeared to be 'terminal-like' swellings which closely apposed retrogradely labeled preganglionic perikarya and proximal dendrites. GLY-LIR somal and dendritic processes were intermingled among retrogradely labeled preganglionic neurons in the nucleus of Terni. None of these GLY-LIR cells were retrogradely labeled. The neuropils surrounding sympathetic preganglionic neurons in the rat also contained numerous GLY-LIR puncta; there were, however, qualitative differences in the density of such profiles across the preganglionic subnuclei. Within the central autonomic and intercalated regions there were numerous GLY-LIR processes, many of which closely apposed retrogradely labeled sympathetic preganglionic somas and proximal dendrites. Within the principal preganglionic cell column, the nucleus intermediolateralis pars principalis (Ilp), there were very few GLY-LIR 'terminal-like' swellings closely apposed to cell bodies in regions of high somal packing density. In regions were this density diminished, GLY-LIR puncta closely apposed retrogradely labeled perikarya and proximal dendritic processes. GLY-LIR spinal neurons were never observed to be within Ilp proper but were present in areas immediately dorsal (lateral lamina V), medial and ventral (lateral lamina VII). GLY-LIR neurons were never retrogradely labeled. The ultrastructural features of GLY-LIR terminals within the sympathetic preganglionic neuropils of both vertebrates were nearly identical. GLY-LIR terminal boutons formed synaptic contacts with retrogradely labeled preganglionic somas as well as with large and medium-sized proximal dendrites. The majority of identified GLY-LIR terminals, however, contacted non-retrogradely labeled medium and small caliber dendrites within the preganglionic neuropils. Ninety-eight percent of GLY-LIR synapses formed symmetric specializations with the postsynaptic element. Ninety-six percent of the GLY-LIR terminal boutons contained some combination of pleomorphic vesicles. These light and electron microscopic observations support the hypothesis that glycine is localized in terminals presynaptic to sympathetic preganglionic perikarya and dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)