Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP), Calcitonin Gene-Related Peptide (CGRP) and Vasoactive intestinal peptide (VIP) are multi-functional neuropeptides that have been linked to debilitating conditions such as migraine. Moreover, Anti-PACAP and anti-CGRP antibodies have demonstrated clinical efficacy in Phase II and Phase III trials for treatment of migraine, respectively. Due to considerable homology and structural similarities, VIP and PACAP can bind to and signal via the same three functional G-Protein Coupled Receptors (GPCR), PAC1R, VPAC1R and VPAC2R. CGRP on the other hand acts via a different mechanism but displays some functional similarities with VIP and PACAP. While both VIP and PACAP can signal through the same receptors, suggesting they may act functionally redundant, differences in receptor affinity, receptor expression levels and biodistribution of each neuropeptide are likely to confer ligand- and cell-type specific signaling. Building on this foundation, our research explores the potential synergies and interplays between neuropeptides such as CGRP, PACAP and VIP, leveraging a variety of in vitro cellular models. Through these studies we aim to contribute to the broader understanding of the various neuropeptide signaling systems and their crosstalk to improve our understanding of the complexities of these interactions and their implications for potential new therapeutic interventions.