Inability of Ehrlich ascites tumor cells to volume regulate following a hyperosmotic challenge

Ehrlich cells shrink when the osmolality of the suspending medium is increased and behave, at least initially, as osmometers. Subsequent behavior depends on the nature of the hyperosmotic solute but in no case did the cells exhibit regulatory volume increase. With hyperosmotic NaCl an osmometric response was found and the resultant volume maintained relatively constant. Continuous shrinkage was observed, however, with sucrose-induced hyperosmolality. In both cases increasing osmolality from 300 to 500 mOSM initiated significant changes in cellular electrolyte content, as well as intracellular pH. This was brought about by activation of the Na+/H+ exchanger, the Na/K pump, the Na+ + K+ + 2Cl cotransporter and by loss of K+ via a Ba-sensitive pathway. The cotransporter in response to elevated [Cl-]i (approximately 100 mM) and/or the increase in the outwardly directed gradient of chemical potential for Na+, K+ and Cl-, mediated net loss of ions which accounted for cell shrinkage in the sucrose-containing medium. In hyperosmotic NaCl, however, the net Cl- flux was almost zero suggesting minimal net cotransport activity. We conclude that volume stability following cell shrinkage depends on the transmembrane gradient of chemical potential for [Na+ + K+ + Cl-], as well as the ratio of intra- to extracellular [Cl-]. Both factors appear to influence the activity of the cotransport pathway.