Neuronal activity is associated with transmembrane ionic redistribution, which can lead to an osmotic imbalance. Accordingly, activity-dependent changes of the membrane potential are sometimes accompanied by changes in intracellular and/or extracellular volume. Experimental data that include distributions of ions and volume during neuronal activity are rare and rather inconsistent partly due to the technical difficulty of performing such measurements. However, progress in understanding the interrelations among ions, voltage and volume has been achieved recently by computational modelling, particularly charge-difference modelling. In this work a charge-difference computational model was used for further understanding of the specific roles for cations and anions. Our simulations show that without anion conductances the transmembrane movements of cations are always osmotically balanced, regardless of the stoichiometry of the pump or the ratio of Na+ and K+ conductances. Yet any changes in cation conductance or pump activity are associated with changes of the membrane potential, even when a hypothetically electroneutral pump is used in calculations and K+ and Na+ conductances are equal. On the other hand, when a Cl- conductance is present, the only way to keep the Cl(-)equilibrium potential in accordance with the changed membrane potential is to adjust cell volume. Importantly, this voltage-evoked Cl--dependent volume change does not affect intracellular cation concentrations or the amount of energy that is necessary to support the system. Taking other factors into consideration (i.e. the presence of internal impermeant poly-anions, the activity of cation-Cl- cotransporters, and the buildup of intra- and extracellular osmolytes, both charged and electroneutral) adds complexity, but does not change the main principles. Author summary We have developed software that calculates membrane potential and cell volume that result from redistribution of principal ions (K+, Na+, and Cl-) during normal cellular activity and experimental manipulations. Calculations in the model are done by an iterative charge-difference method that makes few assumptions about governing equations. Most of the features that were considered to be important for volume and voltage regulation were incorporated in the model, including the unique capability to perform calculations with different values of transmembrane water permeability. We have used the program to reexamine interactions between ionic fluxes, membrane potential, and cell volume and found that there was a previously unappreciated difference in the way that the distribution of cations and anions affect the cell. Na+ and K+, which are distributed unevenly across the membrane by the Na+/K+-ATPase, are primarily responsible for the membrane potential, but, contrary to popular belief, do not directly participate in volume regulation. On the other hand, the Cl- conductance determines the extent of volume changes, because Cl- has to follow the changes of membrane potential, which inevitably leads to changes in cell volume. The software is available to download and use for other investigations.
