Volume regulation by Amphiuma red blood cells: Cytosolic free Ca and alkali metal-H exchange

Peter M Cala, L. J. Mandel, E. Murphy

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

Osmotic swelling of Amphiuma red blood cells results in activation of electroneutral K-H exchange, whereas cell shrinkage activates an electroneutral Na-H exchange. These K-H and Na-H exchanges function to restore cell volume to normal after cell swelling and shrinkage, respectively. Our previous studies have suggested that Ca plays a role in volume-dependent activation of K-H exchange. In the present studies, intracellular free Ca levels were measured employing the Ca-sensitive extracellular dye arsenazo III and a previously described null-point method. Control values for intracellular free Ca averaged 0.46 ± 0.15 μM. Cell shrinkage caused this value to decrease to 0.16 ± 0.11 μM, whereas either cell swelling or addition of 5 μM A23187 resulted in saturation of intracellular Ca buffers, suggesting that both treatments caused an increase in intracellular free Ca. In the presence of 7 μM A23187, the rate of K-H exchange displayed a hyperbolic relationship as a function of extracellular Ca (Ca(o)). The appearent half-maximal concentration for Ca(o) (in the presence of 7 μM A23187) was 0.27 mM for osmotically swollen cells and 1.9 mM for cells in isotonic medium, suggesting that the Ca affinity of a modulating site is increased in swollen cells. Inhibitors of Ca-mediated processes, such as quinidine and the phenothiazines, inhibited K-H exchange. In contrast, the phenothiazines chlorpromazine and trifluoperazine stimulated Na-H exchange by osmotically shrunken cells. These results suggest that increases in intracellular free Ca are involved in stimulating K-H exchange while representing Na-H exchange in Amphiuma red blood cells.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume250
Issue number3
StatePublished - 1986

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

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