Summary
Visceral pain and its affective component associated with cystitis remain poorly understood. Here, we delineate the role of central amygdala (CeA) neuronal ensembles in encoding and modulating cystitis-induced bladder pain and its affective components. Utilizing a multidisciplinary approach combining behavioral assays, optogenetic manipulations, whole-cell electrophysiology, and activity-dependent genetic labeling, we identified functionally discrete CeA subpopulations that are selectively recruited during bladder inflammation. Bidirectional optogenetic modulation of these ensembles produced opposing effects on nocifensive and anxiety-like behaviors, indicating their causal involvement. Single-nucleus RNA sequencing of FosTRAP-labeled neurons revealed distinct transcriptional signatures associated with inflammatory activation. Integrating electrophysiological and transcriptomic data, we demonstrate that cystitis drives cell type–specific adaptations in CeA circuits. These findings provide mechanistic insight and uncover a molecularly and functionally defined CeA ensemble that orchestrates the sensory and affective dimensions of visceral pain.