Cell volume regulation following hypotonic shock in hepatocytes isolated from Sparus aurata

Abstract

The response of isolated hepatocytes of Sparus aurata to hypotonic shock was studied by the aid of videometric and light scattering methods. The isolated cells exposed to a rapid change (from 370 to 260 mOsm/kg) of the osmolarity of the bathing solution swelled but thereafter underwent a decrease of cell volume tending to recovery the original size. This homeostatic response RVD (regulatory volume decrease) was inhibited in the absence of extracellular Ca 2+ and in the presence of TMB8, an inhibitor of Ca 2+ release from intracellular stores. It is likely that Ca 2+ entry through verapamil sensitive Ca 2+-channels, probably leading to a release of Ca 2+ from intracellular stores, is responsible for RVD since the blocker impaired the ability of the cell to recover its volume after the hypotonic shock. RVD tests performed in the presence of various inhibitors of different transport mechanisms, such as BaCl 2, quinine, glybenclamide and bumetanide as well as in the presence of a KCl activator, NEM, led us to suggest that the recovery of cell volume in hypotonic solution is accomplished by an efflux of K + and Cl − through conductive pathways paralleled by the operation of the KCl cotransport, followed by an obliged water efflux from the cells.