Isovolumetric regulation of C6 rat glioma cells in hyperosmotic media

Volume regulation of C6 glioma cells was studied in response to a gradual increase of extracellular osmolality from 300 to 440 mosmol/kgH2O at 37 degrees C. Maintenance of cell size depended on the rate of osmolality increase (CR): at CR of 3 mosmol.kg-1.min-1, cell volume was kept constant, whereas it decreased progressively at CR of 6 or 9 mosmol.kg-1.min-1. The ability of C6 cells to maintain their volume is termed isovolumetric regulation (IVR). Reducing temperature to 22 degrees C inhibited IVR significantly. Also, bumetanide and ouabain blocked the regulation, while 5-(N,N-dimethyl)amiloride (DMA) did not affect IVR: Extracellular acidification rate (EAR) was studied by microphysiometry. EAR gradually decreased in the presence and increased in the absence of IVR. Experiments with DMA show that these changes in EAR were related to the activity of the Na+/H+ exchanger. It was stimulated by cell shrinkage but not by hyperosmolality itself. Our data demonstrate that C6 glioma cells are able to prevent volume decrease at a low rate of elevation of external osmolality and at 37 degrees C. This process requires electrolyte uptake by the Na(+)-K(+)-2Cl cotransporter and Na+/K+ pump.