Axotomy of the rat superior cervical ganglion results in a two-fold increase of neuropeptide tyrosine as determined by radioimmunoassay. On the other hand, treatment of sympathetic neuron cultures with leukemia inhibitory factor, a cytokine that is known to be involved in the up-regulation of galanin after axotomy in vivo, decreases neuropeptide tyrosine messenger RNA. These, apparently contradictory findings, prompted us to investigate the regulation of neuropeptide tyrosine in the axotomized superior cervical ganglion in vivo. For comparison, the regulation of galanin was examined under the same conditions. Compared to control ganglia, the number of neuropeptide tyrosine-positive cell bodies decreased while the density of immunoreactive neuronal processes increased one week after transection of the major postganglionic nerves. The nerve fibres were identified as axons by the absence of MAP2, a somatodendritic marker protein. They extended into both carotid nerves and ramified at the lesion site. In situ hybridization revealed that, although the number of neuropeptide tyrosine messenger RNA-positive neurons was not different from controls, the average grain density/neuron decreased by 40%. When axotomized ganglia were decentralized simultaneously, a three-fold elevation of neuropeptide tyrosine immunoreactivity was detectable by radioimmunoassay and an additional increase in numerical density of neuropeptide tyrosine-immunoreactive nerve fibres was observed. Levels of neuropeptide tyrosine messenger RNA were significantly reduced within postganglionic neurons. This synergistic effect of combined axotomy and decentralization on peptide content was also detected for the neuropeptide galanin that, in contrast to neuropeptide tyrosine, is induced by axotomy or decentralization on protein and messenger RNA level. Therefore, while neuropeptide tyrosine messenger RNA is reduced in axotomized ganglia (most likely in response to leukemia inhibitory factor), the peptide accumulates in axonal processes resulting in increased peptide levels as determined by radioimmunoassay.