The influence of chloride ions on renal outer medullary cell volume

1. Slices of rat renal outer medulla have been incubated in media made hyperosmotic (540 and 1055 m-osmole/kg H2O) by the addition of urea, and containing variable concentrations of Cl (90, 144, 189 and 215 mM) and constant concentrations of Na (180 mM) and K (5.9 mM). A small number of incubations have been conducted in the presence of 100 mM-Na. 2. Changes in cell volume during incubation have been calculated on the basis of initial and final slice weight and inulin space. 3. The capacity of cells to shrink in response to extracellular osmotic stress was related principally to the external Cl concentration rather than to osmolality, increases in concentration being associated with enhanced shrinkage. Shrinkage was accompanied by net loss of cellular Cl. The ratio between intra- and extracellular Cl concentration (ca. 0.41) remained constant in all media. 4. In media containing low Cl concentration (90 mM), reduction of media Na concentration to 100 mM enhanced shrinkage. This effect was not observed when medium Cl concentration was greater than 90 mM. 5. Ethacrynic acid-cysteine (1 mM) significantly impaired the shrinkage response to extracellular osmotic stress, and caused cell swelling in media of relatively low Cl concentration and osmolality. It did not abolish the dependency of cell volume upon Cl concentration. There was marked reduction in the net amount of Cl lost from cells. 6. Ethacrynic acid-cysteine caused an increase in cellular Na content only in media containing 540 m-osmole/kg H2O and Cl concentrations less than 215 mM. 7. Ouabain (1 mM) inhibited cell shrinkage to a lesser extent than ethacrynic acid-cysteine in all media except that causing the greatest shrinkage under control conditions (215 mM-Cl/1055 m-osmole/kg H2O). It is suggested that a ouabain-sensitive process may play an increasingly important role in Cl-related cell shrinkage as this becomes more pronounced. 8. The findings are consistent with the view that Cl ions influence cell volume both through their effective external osmotic pressure and by means of Cl-specific process; the latter is associated with net loss of cellular Cl. A dependence of this loss upon Na/k exchange-linked metabolism is inferred, but the present findings do not permit the active or passive nature of the extrusion to be defined.