Erythrocytes possess a Cl-dependent, Na-independent K transport system cotransporting K and Cl in a 1:1 stoichiometry that is membrane potential independent. This K-Cl cotransporter is stimulated by cell swelling, acidification, Mg depletion, and thiol modification. Cell shrinkage, elevation of cellular divalent ions, thiol alkylation, phosphatase inhibitors, and derivatives of certain loop diuretics and stilbenes are inhibitory. Thus regulation of K-Cl cotransport at the membrane and cytoplasmic levels is highly complex. Basal K-Cl cotransport decreases with cellular maturation, whereas its modes of stimulation and inhibition are variable between species. The physiological inactivation appears to be prevented in low-K animal erythrocytes. In certain human hemoglobinopathies, K-Cl cotransport may be the cause of cellular dehydration and volume decrease. K-Cl cotransport occurs also in nonerythroid cells, such as in epithelial and liver cells of other species. At the threshold of molecular characterization, this comprehensive review places our present understanding of the mechanisms modulating K-Cl cotransport physiologically and pathophysiologically into kinetic and thermodynamic perspectives.

K-Cl cotransport (COT), defined first in red blood cells as the Cl-dependent, ouabain-insensitive bidirectional K transport, encoded by at least four KCC (kalium-chloride-cotransport) genes, is now recognized as a functional and structural reality in all cell membranes. As functional system, K-Cl COT is necessary for volume and ionic homeostasis. Since its original discovery by swelling red cells in hyposmotic solutions and by treatment with N-ethylmaleimide (NEM), K-Cl COT has been recognized as one of the prime electroneutral, low ion affinity pathways effecting regulatory volume decrease (RVD). This review first summarizes the general properties of K-Cl COT, including ion dependence, kinetics, thermodynamics and regulation in erythrocytes of various species, and then focuses on the newest findings of the molecular mechanisms behind K-Cl COT, the KCC isoforms and their expression in epithelial cells and in Xenopus oocytes. Based on early biophysical studies on red cells amalgamated with the recent molecular expression studies of the four KCC isoforms, K-Cl COT emerges as one of the oldest membrane transporters that is controlled by a complex redox-dependent cascade of kinases and phosphatases, yet to be defined at the molecular level. Whereas RVD is a primeval role of K-Cl COT for survival of cells challenged by hyposmotic environments, maintenance of intracellular Cl ([Cl]^I ) levels away from electrochemical equilibrium and K buffering capability during neuronal function are new additions to the list of physiological functions of this system.

Treatment with 2 mM N-ethylmaleimide (NEM) caused a marked increase in K+ permeability of low K+ but not of high K+ sheep red cells suspended in isosmotic Cl- media with 10(-4) M ouabain. The Na+ permeability was unaltered. Kinetic analysis by K+ efflux and K+ or Rb+ influx measurements suggests that NEM primarily increased the bidirectional fluxes of K+ and Rb+, since (a) no significant change in the apparent external affinities of these ions was found, and (b) below unity, the observed flux ratios were close to those calculated from the Ussing relationship. Replacement of Cl- by NO3 abolished the NEM-stimulated and reduced the basal K+ flux rates. Similarly, 10(-3) M furosemide inhibited Cl- -dependent K+ fluxes in both control and NEM-treated LK red cells. Exposure of LK cells to hyposmotic but not to hyperosmotic salt solutions increased the basal Cl- dependent K+ flux twofold as reported by Dunham and Ellory (J. Physiol. (London) 318:511-530, 1981) but did not affect its fractional stimulation by NEM. The action of NEM is interpreted as a stimulation of a temperature-dependent and Cl- -requiring K+ transport pathway genetically preserved in adult LK but turned off in HK sheep red cells. In addition, common to both LK and HK sheep red cells was a basal K+ flux that operated in the presence of either Cl- or NO3-.