[3H]ouabain binding during the monolayer organization and cell cycle in MDCK cells

Abstract

The specific binding of [3H]ouabain in an epithelial cell line derived from a dog kidney (MDCK) was determined during epithelial reorganization and also during the cell cycle. In suspended cells, the specific binding of [3H]ouabain is reduced 67% compared with the binding obtained in a complete monolayer. After plating back these cells on a permeable support, both transepithelial electrical resistance and [3H]ouabain binding increase with time of incubation. [3H]ouabain binding decreases during S and G2 phases of the cell cycle to reach a minimum during mitosis and increases again during GI. The transepithelial electrical resistance, determined simultaneously, shows the same behavior. The reduction in the number of [3H]ouabain binding sites in two different circumstances in which the epithelial membrane organization is disrupted and the increase in [3H]ouabain binding sites when the epithelial membrane is reorganized are consistent with the hypothesis that the number of pumping sites responsible for the active step in the transepithelial active transport is additional to the number required to maintain the intracellular ionic composition.