Preganglionic and sensory axons in developing bullfrog sympathetic ganglia express three neuropeptides during early...

Retrograde tracing in fixed tissue with the fluorescent carbocyanine dye, DiI, was used to identify neurons that project to paravertebral sympathetic ganglia 9 and 10 in bullfrog tadpoles. Applying DiI to ganglion 9 at stage II labelled spinal preganglionic neurons and sensory neurons in dorsal root ganglia. When examined in a stage V tadpole, the segmental boundaries of the preganglionic cell column which supply the lumbar chain ganglia were identical to that in the adult. Using immunocytochemistry, luteinizing hormone releasing hormone-like immunoreactivity, calcitonin gene-related peptide-like immunoreactivity, and substance P-like immunoreactivity were localized at stage III in axons within sympathetic ganglia 9 and 10. During subsequent stages, the density of fibers containing these peptides increased and it became easier to identify synaptic boutons in contact with postganglionic neurons. These observations demonstrate that projections to the lumbar sympathetic ganglia are already formed by the earliest tadpole stages, they are consistent with the previous physiological observation of nicotinic synapses in stage III ganglia, and they suggest that neuropeptide function may also begin during early stages.