Effects of anisosmotic medium on cell volume, transmembrane potential and intracellular K+ activity in mouse...

Mouse hepatocytes in primary monolayer culture (4 hr) were exposed for 10 min at 37 degrees C to anisosmotic medium of altered NaCl concentration. Hepatocytes maintained constant relative cell volume (experimental volume/control volume) as a function of external medium relative osmolality (control mOsm/experimental mOsm) ranging from 0.8 to 1.5. In contrast, the relative cell volume fit a predicted Boyle-Van't Hoff plot when the experiment was done at 4 degrees C. Mouse liver slices were used for electrophysiologic studies, in which hepatocyte transmembrane potential (Vm) and intracellular K+ activity (aik) were recorded continuously by open-tip and liquid ion-exchanger ion-sensitive glass microelectrodes, respectively. Liver slices were superfused with control and then with anisosmotic medium of altered NaCl concentration. Vm increased (hyperpolarized) with hypoosmotic medium and decreased (depolarized) with hyperosmotic medium, and in [10(experimental Vm/control Vm)] was a linear function of relative osmolality (control mOsm/experimental mOsm) in the range 0.8-1.5. The aik did not change when medium osmolality was decreased 40-70 mOsm from control of 280 mOsm. Similar hypoosmotic stress in the presence of either 60 mM K+ or 1 mM quinine HCl or at 27 degrees C resulted in no change in Vm compared with a 20-mV increase in Vm without the added agents or at 37 degrees C. We conclude that mouse hepatocytes maintain their volume and aik in response to anisosmotic medium; however, Vm behaves as an osmometer under these conditions. Also, increases in Vm by hypoosmotic stress were abolished by conditions or agents that inhibit K+ conductance.