The ionic basis of the membrane potential in a rat glial cell line

Intracellular ionic concentrations, membrane potential and Na+ and K+ flux were measured in a clonal rat glial cell line, C6. Intracellular concentrations of C6 cells were: (mmole/liter cell water) K+ 145 +/- 4 S.D., Na+ 18 +/- 4 S.D., Cl- 14 +/- 1 S.D. Cells maintained a steady state level of K+ over the duration of the experiments. This was substantiated by the close agreement between absolute values for K+ influx and efflux measured with 42K. When cells were depleted of internal Na+, K+ influx was significantly reduced suggesting that a portion of inward K+ movement is linked to Na+ extrusion. Efflux of Na+, calculated from the half-time of exchange from cells preloaded with 22Na, was higher than passive Na+ influx determined by blocking Na+ extrusion with ouabain. Since cell Na+ concentration remained relatively constant, part of the Na+ efflux may be due to exchange diffusion. The average membrane potential of C6 cells uas -36 mV. The potential showed a 31 mV slope for a 10-fold change in external K+ so that it is determined predominantly by the ratio of external/internal K+. The potential however, consistent with the relatively large passive Na+ influx, was influenced by Na+. Replacing all but 20 mM external Na+ with choline hyperpolarized the membrane by 13 mV. The ratio of PNa/PK of 0.11 suggests that Na+ is outwardly transported to maintain the steady-state concentrations observed. Since these responses are similar to those in non-tumor glia, it suggests that the C6 cell line may provide a useful model for studying ionic regulation in glial cells.