Calcitonin gene-related peptide (CGRP) has been found in both sensory and motor neurons. It has been suggested that CGRP is transported from neuron cell bodies to their terminals, where it may act as an anterograde trophic factor. However, it is not known how fast CGRP is transported or whether CGRP found in the innervated target organ indeed originated in neural tissues. We have quantified endogenous CGRP in the rat peripheral nerve by a newly developed enzyme immunoassay. The CGRP immunoreactive material obtained from neural tissues coincided with synthetic rat CGRP in fractional distributions separated by gel filtration. After ligation of the sciatic nerve, tissue CGRP accumulated in the segment central to the ligature. The rate of anterograde transport of CGRP was about 1 mm/hr in both sensory and motor fibers. In the sciatic nerve, only a small fraction of CGRP measured was found to originate from the motor nerve fibers. This may be due in part to the disproportionately large number of sensory fibers in the sciatic nerve and in part to the possible presence of CGRP in sympathetic nerve fibers. The CGRP content in the dorsal root fibers was significantly lower than that in the peripheral processes of the sensory neurons. The CGRP content in the hind leg muscle was much higher than that expected from the amount of CGRP per nerve fiber in the sciatic nerve. Most CGRP in muscle disappeared following denervation. It is concluded that CGRP highly concentrated in nerve terminals is supplied by axonal transport from the neuron cell bodies.