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Embryonic expression of pituitary adenylate cyclase‐activating polypeptide in sensory and autonomic ganglia and in spinal cord of the rat

Pituitary adenylate cyclase‐activating polypeptide (PACAP), a neuropeptide that is related structurally to vasoactive intestinal polypeptide (VIP), has been shown to stimulate neuronal growth and differentiation, indicating a possible function in the development of the nervous system. Studies have indicated that the PACAP receptor is expressed during development, but data on PACAP expression are limited mainly to postnatal development. In the present study, we used immunohistochemistry and in situ hybridization histochemistry to examine the expression of PACAP in autonomic and sensory ganglia and spinal cord of rat fetuses at embryonic days 12–21 (E12–E21). PACAP immunoreactivity was visualized by using a specific monoclonal anti‐PACAP antibody to detect both PACAP‐38 and PACAP‐27, and PACAP mRNA was visualized by using a [33P]‐labeled cRNA‐probe. PACAP+ nerve fibers were observed in the spinal cord as early as E13. At E14, PACAP‐immunoreactive nerve fibers projected to the sympathetic trunk, where few PACAP+ nerve cell bodies were seen from E15. On the same embryonic day, PACAP‐immunoreactive nerve cell bodies appeared in the intermediolateral column of the spinal cord. From E15 to E16, PACAP‐immunoreactive nerve cell bodies were visible within sensory and autonomic ganglia, such as the dorsal root, the trigeminal, the sphenopalatine, the otic, the submandibular, and the nodose ganglia. At E16, PACAP+ nerve fibers were innervating the adrenal medulla, and immunoreactive fibers could also be observed in the superior cervical ganglion, in which PACAP‐immunoreactive cell bodies were detected occasionally from E18. The synthesis of PACAP in neuronal cell bodies was confirmed by the demonstration of PACAP mRNA with in situ hybridization histochemistry.