The rat esophagus shares some cellular features with skin squamous epithelium and striated muscle that express high levels of CRF₂ receptors or their cognate ligand urocortins (Ucn). We investigated the expression and cell signaling of CRF₂ receptors and ligands in the rat esophagus and lower esophageal sphincter (LES) by reverse transcriptase-PCR and quantitative PCR in normal and corticosterone-treated whole esophageal tissue, laser capture microdissected layers, and isolated esophageal cells. The expression of CRF₂ receptor protein and intracellular cAMP and ERK1/2 responses to CRF agonists and CRF₂ antagonist were determined in cultured esophageal cells and HEK-293 cells transfected with CRF_2b receptors. CRF₂ was abundantly expressed in the mucosa and longitudinal muscle layers of the esophagus and LES while CRF₁ expression was scarce. CRF_2b wild-type transcript was predominantly expressed in the esophagus and, in addition, several new CRF₂ splice variants including six CRF_2a isoforms were identified. Expression of Ucn 1, Ucn 2 and to a smaller extent, Ucn 3, but not CRF mRNA was detected in the esophagus and LES. Ucn 1 and Ucn 2 stimulated dose-dependent cAMP production and ERK1/2 phosphorylation in the esophageal cells, while CRF and CRF₁ agonist, cortagine, had less potent effects. In addition, Ucn 2-stimulated cAMP and ERK responses were blocked by the CRF₂ antagonist, astressin₂-B. These data established the presence of a prominent CRF₂ signaling system in the esophagus and LES encompassing multiple CRF₂ receptor variants and urocortins, suggesting a functional role in secreto-motor activity, and epithelial and muscle cell proliferation.