Characterization of voltage-dependent calcium influx in human erythrocytes by fura-2

Biochem Biophys Res Commun. 1997 Jul 30;236(3):549-54. doi: 10.1006/bbrc.1997.7002.

Abstract

Thus far, the methods used to determine erythrocyte Ca2+ influx have not allowed the assessment of the kinetics of ion uptake. To overcome this drawback, we studied a new method, using the fluorescent Ca2+-chelator fura-2, which directly quantifies intracellular Ca2+ changes in human erythrocytes. This method has the advantage over previous techniques that it monitors continuously cellular Ca2+ levels. The Ca2+ influx is modulated by cellular membrane potential in the presence of a transmembrane Ca2+ concentration gradient and exhibits a first slow increase of the intracellular Ca2+ concentration, followed, after the reachment of a threshold value of 125 +/- 13 nM Ca2+, by a faster increase until a plateau is reached. The influx rate is inhibited by dihydropyridines in the micromolar range. These findings support the hypothesis that erythrocyte Ca2+ influx is mediated by a carrier similar to the slow Ca2+ channels and is dependent on membrane depolarization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / pharmacology
  • Calcium / blood*
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels / drug effects
  • Calcium Channels / physiology
  • Dihydropyridines / pharmacology
  • Electrochemistry
  • Erythrocytes / metabolism*
  • Fluorescent Dyes*
  • Fura-2*
  • Humans
  • Membrane Potentials
  • Potassium / pharmacology
  • Temperature

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Dihydropyridines
  • Fluorescent Dyes
  • 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
  • Potassium
  • Calcium
  • Fura-2