Dynamic NMR measurement of volume regulatory changes in Amphiuma RBC Na+ content

S. E. Anderson, J. S. Adorante, Peter M Cala

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

23Na nuclear magnetic resonance (NMR) and conventional chemical methods were employed to measure Na+ fluxes in Amphiuma red blood cells (RBC) during volume regulation. Paramagnetic shift reagents [dysprosium triethylenetetraminehexaacetic adid (DyTTHA) and dysprosium tripolyphosphate (Dy(TPP)2)] were used to alter extracellular Na+ magnetic resonance. Data are presented describing 23Na resonance dependence on shift reagent, sodium and calcium concentration. We confirmed that the shift reagents neither enter the cells nor alter intracellular Na+, K+, and Cl- concentrations under control conditions when extracellular calcium was maintained > 0.5 mM. We also confirmed that the shift reagent complexes chelate calcium [Dy(TPP)2 much more so than DyTTHA] and that their toxic effects could be alleviated by adjusting calcium in the cell's suspension medium to control levels. In parallel experiments, where volume-activated Na+ fluxes ranged from 0.3 to 3 mmol Na+/kg dry cell solid (DCS) x minute in cells containing from 30 to 150 mmol Na+/kg DCS, changes in intracellular sodium measured by 32Na NMR were within 4% of those measured by conventional destructive methods. Finally, we present data that are consistent with the interpretation that 6 mmol Na+/kg DCS plus 16% of intracellular Na+ is NMR invisible.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume254
Issue number3
StatePublished - 1988

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

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