PMID: 3094375Oct 1, 1986

Nuclear magnetic resonance measurement of cytosolic free calcium levels in human red blood cells

The American Journal of Physiology
E MurphyR E London

Abstract

Red blood cells were loaded with 1,2-bis(2-amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid (FBAPTA) by incubation with 50 microM of the acetoxymethyl ester (FBAPTA-AM), and cytosolic free Ca2+ was monitored with 19F-nuclear magnetic resonance (NMR). Loading with 50 microM FBAPTA-AM, which results in a final FBAPTA level of approximately 0.5 mM, caused only a 25-30% fall in cell ATP as measured by 31P-NMR when 5 mM pyruvate was present. Leakage of the NMR active Ca2+ indicator, which results from cell lysis, was corrected for with the addition of extracellular Eu3+ ions, extracellular ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA), or washing. With this method, we have found basal levels of cytosolic free Ca2+ averaging 61 +/- 6 nM (means +/- SE, n = 19). When the intracellular level of FBAPTA was varied from 0.1 to 1.0 mM, there was no correlation between the level of cytosolic free Ca2+ and the level of loading with FBAPTA. Addition of 10 microM of the Ca2+ ionophore A23187 with extracellular Ca2+ set at different levels by Ca2+-EGTA buffers caused an increase in cytosolic free Ca2+ as expected. Furthermore, ATP depletion caused a two- to three-fold increase in cytosolic free Ca2+, consistent wit...Continue Reading

Citations

Jan 1, 1987·The Journal of Membrane Biology·B Engelmann, J Duhm
Sep 13, 2000·Journal of the American Geriatrics Society·M BarbagalloL M Resnick
Aug 1, 1992·Pflügers Archiv : European journal of physiology·B EngelmannJ Duhm
Sep 1, 1987·European Journal of Biochemistry·E FernandezP Laszlo
Jul 12, 2003·High Altitude Medicine & Biology·Margueritte M White, Lubo Zhang
Dec 13, 2005·European Biophysics Journal : EBJ·Anthony D MaherP W Kuchel
Mar 1, 1990·Environmental Health Perspectives·F A SchanneJ F Rosen
Dec 31, 1997·American Journal of Respiratory and Critical Care Medicine·M Toporsian, M E Ward
Mar 1, 1990·Environmental Health Perspectives·E MurphyR E London
Jan 18, 2019·BMC Genomics·Liana ApolisSanjay A Desai

Related Concepts

5,5'-difluoro-1,2-bis(2-aminophenoxy)ethane-N, N, N', N'-tetraacetic acid
Adenosine Triphosphate, Chromium Ammonium Salt
Calcium
Cytoplasmic Matrix
Egtazic Acid Sodium Salt
Erythrocytes
In Vivo NMR Spectroscopy

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