Vasoactive-intestinal-polypeptide (VIP) is a neuropeptide that is localized together with its receptors (VPAC1 and VPAC2) in brain areas of the stress and anxiety circuit, such as central amygdala (CeA). However, the exact role of the CeA-VIP system in stress and anxiety function is not fully known. By using a pharmacological and chemogenetic approach we investigated the role of VIP in the CeA on anxiety and stress responses of mice. First, we administered specific VIP/VPAC receptor agonists/antagonists bilaterally into the CeA to evaluate effects on anxiety- and stress-related behaviors as well as neuroendocrine stress response. We found that intra-CeA infusions of VIP cause anxiogenic-like behavioral effects, reduced risk-assessment in behavioral tasks for the assessment of anxiety-related behavior such as elevated plus-maze and a more passive coping style in the forced swim test, compared to vehicle-injected controls. Conversely, intra-CeA administration of VPAC1/2 receptor antagonists induced opposite behavioral changes suggesting anxiolytic-like efficacy and improved stress coping ability. Moreover, in a subsequent experiment we chemogenetically activated VIP+-neurons within the ventral periaqueductal grey (vPAG) the main source of VIP innervation of the CeA through cre-dependent designer-receptors-exclusively-activated-by-designer-drugs (DREADDs) to reveal the impact of a PAG-VIP-CeA circuit in stress- and anxiety function. Our data show that CNO-induced activation of VIP+-neurons of the vPAG resulted in an increase of anxiogenic-like behavior and reduced risk-assessment in the elevated plus-maze and light-dark test. Thus, our findings identify a VIP-mediated circuit in the brain that plays a critical role in the regulation of stress- and anxiety-related behaviors indicating translational potential.(FWF-P33534-B)