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Little is known of how the presence of cranial autonomic symptoms (CAS) in cluster headache and migraine may contribute to their severe headache phenotype. However, this strong association points to the involvement of the cranial parasympathetic efferent pathway. To dissect its contribution, we investigated the role of pituitary adenylate cyclase-activating polypeptide-38 (PACAP38), a potent sensory and parasympathetic neuropeptide, in modulating pre- and post-ganglionic cranial parasympathetic projection neurons, and their influence on headache-related trigeminal-autonomic responses.