Relationships between morphological and electrophysiological changes with low concentrations of ethanol on in vitro guinea pig gastric mucosa were investigated. Tissues mounted in Ussing chambers allowed recording of transepithelial potential difference (PD), resistance (R), short-circuit current (Isc), and acid secretion (H+). At selected times the mucosae were processed for morphological analysis. With luminal 10% ethanol there was a decrease in PD, R, Isc, and H+ within 1 min, and they eventually went to low steady-state values between 10 and 40 min. At 1 min many surface epithelial cells lifted off from the basal lamina but were still anchored by thin basal cell processes. After 10 min in ethanol many surface cells had completely detached from the basal lamina but remained connected to adjacent cells by their junctions. Numerous cytoplasmic blebs formed on both apical and basal cell surfaces. Concurrently, there was a significant increase in microvillus length. After 40 min most of the surface cells were detached from the basal lamina as sheets forming epithelial blisters. Upon ethanol washout there was epithelial cell reattachment to the basal lamina and a return of the PD, R, and Isc to control values within 40 min. Incubation of the luminal surface with 10% ethanol for 5 h resulted in a gradual rise of the PD, R, Isc, and H+ to control values by 4 h with a coincident return of the normal mucosal morphology. These studies indicate that ethanol has reversible and possibly adaptable effects on the in vitro guinea pig gastric mucosa and that the morphological changes are closely correlated with the decline and recovery of the electrical and secretory activity of the tissue.