Studies on the Relationship Between Rat Renal Medullary Cell Volume and External Anion Concentration in Hyperosmolal Media

1. The volumes of cells in slices of rat renal outer medulla have been examined following incubation or 25 min in hyperosmolal media (650 and 950 m-osmole/kg H2O) containing independently variable concentrations of Cl (70-325 mM) and HCO3 (10-60 mM) (gas phase 95% O2/5% CO2). 2. For any given level of external Cl concentration cell volumes were reduced by increasing the external HCO3 concentration. These reductions were accompanied by net loss of cellular K and Cl. In confirmation of earlier findings, cell volumes were also reduced by increasing external Cl concentration. 3. Experiments in which the HCO3 concentration and pH of the incubation media were independently varied by the use of N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid (HEPES)/100% O2 showed that it is the HCO3 anion per se which influences cell volume. 4. The anion exchange inhibitor 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid (SITS, disodium salt, 1 mM) abolished the dependence of cell volume upon HCO3 but not upon Cl. 5. Acetazolamide (1 mM) influenced (reduced) cell volumes only in the presence of low (10 mM) HCO3. 6. CNS (25 mM) also markedly reduced cell volumes in media containing 10mM-HCO3 and, to a lesser extent, 25 mM-HCO3. It was without effect on cell volume when external HCO3 was 60 mM. 7. The presence of CNS was associated with the significant cellular net accumulation of Cl in media in which either Cl or HCO3 concentration (or both) was low (70 or 130 mM and 19 mM respectively). 8. The outer medullary [35S]CNS space at 25 min, determined for slices incubated in a representative selection of the various media employed in this study, exceeded the [14C]inulin space by 1.77 microliters/10 mg wet weight.