The aim of this study was to develop a new cellular restoration model using primary cultured rabbit gastric mucosal cells, and to evaluate the role of cytoskeleton and calmodulin in this process. Confluent monolayer mucosal cell sheets consisting mainly of mucous cells were wounded, and a rotating silicon tip was used to make cell-free areas of constant size. The process of restoration was monitored, and the cell-free area was measured and analysed quantitatively. Artificial wounds recovered in 36 h in controls; however, mucosal cell repair was inhibited by treatment with the actin inhibitor, cytochalasin B, and the calmodulin inhibitor, W-7. In the process of restoration, bromodeoxyuridine (BrdU)-positive cells appeared around the wound 24 h after injury and then disappeared after the complete repair. Because BrdU-positive cells were never detected within 24 h after wounding, initial recovery of the wound occurred only by cell migration. Both the migration and the proliferation stage are seen with this model. This culture model is suitable for the analysis of gastric mucosal restoration. Present data are consistent with the hypothesis that the integrity of the cytoskeletal system is important for restoration after damage of the gastric mucosa.