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Intracellular Na+ and K+ activities have been determined under visual control in enterocytes located at known positions along hamster ileal villi in vitro. The intracellular K+ activity found in enterocytes migrating over the basal third of villi was significantly less than that found in older enterocytes (83 and 113 mM respectively). The intracellular Na+ activity remained unaffected by the state of enterocyte development (32 mM throughout). Extracellular K+ and Na+ activities measured close to the luminal surface of the intestine were always similar to those measured in the bulk of the bathing medium. There was, however, a small difference in K+ activity detected between the bulk solution and solution near the luminal surface of enterocytes (3 . 4 and 3 . 6-3 . 7 mM respectively). Na+ activity in the villus core was also similar to that of the bathing medium. The brush border membrane content of sucrase-isomaltase, determined cytochemically, was found to increase rapidly as enterocytes migrated over the basal third of the villus and to plateau at a point roughly equivalent to that where intracellular K+ activities became maximal. The numerous ways by which enterocytes might control their intracellular K+ concentrations during development are emphasized. Attention is nevertheless drawn to the close correspondence seen between different aspects of brush border membrane development and the ability of enterocytes to maintain high internal concentrations of K+.