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31 P and 23 Na NMR spectroscopy of normal and ischemic rat skeletal muscle. Use of a shift reagent in vivo
Author(s) -
Balschi James A.,
Bittl John A.,
Springer Charles S.,
Ingwall Joanne S.
Publication year - 1990
Publication title -
nmr in biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.278
H-Index - 114
eISSN - 1099-1492
pISSN - 0952-3480
DOI - 10.1002/nbm.1940030202
Subject(s) - extracellular , chemistry , reagent , skeletal muscle , sodium , myoglobin , extracellular fluid , intracellular , in vivo , nuclear magnetic resonance spectroscopy , analytical chemistry (journal) , nuclear magnetic resonance , biochemistry , chromatography , anatomy , stereochemistry , biology , physics , microbiology and biotechnology , organic chemistry
23 Na NMR spectroscopy was used 1, to define the distribution of the shift reagent for cations, triethylenetetraminehexaacetatedysprosium(III), DyTTHA 3− , in the living rat; 2, to define the characteristics of the Na resonances reporting intra‐ and extracellular Na + in skeletal muscle in vivo ; and 3, to calculate the Na + concentrations in the intra‐ and extracellular spaces of the gastrocnemius muscle during well‐perfused and ischemic conditions. The concentration of DyTTHA 3− infused intravenously into the jugular vein of the living rat reached a maximum value of 8–9 m M in the extracellular space of the muscle after ca 40 min of infusion. This allowed excellent discrimination of extra‐ and intracellular Na signals (Na o and Na i , respectively) and did not spoil the resolution of concurrent 31 P NMR spectra. Infusion of shift reagent changed neither hemodynamic performance of the rat nor the high‐energy phosphate content of skeletal muscle. Shift reagent enters ca 15% (v/w) of the rat body weight; this corresponds to almost all of the “fast” or rapidly permeable extracellular space. It is excreted from the body with a pseudo‐first order rate constant of 0.0158 min −1 . In resting muscle, we estimate that [Na + ] i is 3–5 m M and, in muscle perfused with the sodium salt of the shift reagent, that [Na + ] o in the fast exchangeable extracellular space is 166 m M . During 11 h of ischemia at 37°C, the area of the Na i + signal area monotonically increased sixfold. Based on estimates for maximum changes in fluid shifts reported by the decrease in the area of the Na o signal area, we calculate that the lower limit for [Na + ] i after 11 h of ischemia is 27 m M . The NMR‐visibility factors for the extracellular and intracellular Na + signals are essentially the same. This study demonstrates that the shift reagent DyTTHA 3− is acutely non‐toxic and that the 23 Na NMR spectra obtained can be used to quantitate [Na + ] o and [Na + ] i in tissues in vivo . Using this technique, we found that the transmembrane sodium gradient fell from ca 35 in well‐perfused skeletal muscle to <6 during prolonged ischemia.