In this study, we integrated behavioral assays, chemogenetic manipulation, tract-tracing methods, RNAscope in situ hybridization, and 4 projection- and synapse-specific eCB-targeted molecular probes to elucidate the contributions of eCB signaling in AI-PrL and AI-ovBNST circuits to the pathophysiology of comorbid headache and anxiety. Here, PRL is linked to Anxiety.