23Na nuclear magnetic resonance study of intracellular sodium in vascular endothelial cells

Biochim Biophys Acta. 1994 Nov 10;1224(2):228-36. doi: 10.1016/0167-4889(94)90195-3.

Abstract

23Na(+)-NMR was used to determine the intracellular mobility and cell membrane permeability of Na+ in porcine vascular endothelial cells. The cells were grown as monolayers onto microcarrier beads and perfused with a medium containing Dy(P3O10)2(7-) to shift the extracellular from the intracellular Na+ resonance. Using triple quantum coherence filtered NMR experiments and spin echoes, it was shown that not all intracellular Na+ ions are in the extreme narrowing limit. The triple quantum coherence filtered experiments resulted in an observed R2f = 2022 +/- 302 s-1 and R2s = 200 +/- 28 s-1. From spin-echo experiments we obtained R2f = 2200 +/- 355 s-1 and a R2s = 145 +/- 15 s-1. These values are similar to those found in other cell systems and indicate water-Na(+)-protein interactions. Using single quantum NMR, the Na+ permeability of the endothelial membrane was determined. To obtain the Na+ transcellular permeability coefficient the cells were treated with 50 microM ouabain in the perfusion medium. Ouabain inhibits the Na-K pump and caused the intracellular Na+ concentration to increase in time. The permeability coefficient was obtained from the time dependence of the intracellular Na+ concentration. Assuming a monolayer of rectangularly shaped cells, we obtained a value of P = 0.02.10-5 cm s-1.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane Permeability
  • Cells, Cultured
  • Endothelium, Vascular / chemistry*
  • Magnetic Resonance Spectroscopy / methods*
  • Nystatin / pharmacology
  • Ouabain / pharmacology
  • Sodium / chemistry*
  • Sodium / metabolism
  • Sodium-Potassium-Exchanging ATPase / antagonists & inhibitors
  • Swine

Substances

  • Nystatin
  • Ouabain
  • Sodium
  • Sodium-Potassium-Exchanging ATPase