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Department of Internal Medicine, Yale University School of Medicine and Howard Hughes Medical Institute, New Haven, Connecticut 06520-8020
Intracellular glucose concentration in skeletal
muscle of awake rats was determined under conditions of hyperglycemic
(10.2 ± 0.6 mM) hyperinsulinemia (~1,200 pM) and hyperglycemic
(20.8 ± 1.5 mM) hypoinsulinemia (<12 pM) by use of
13C nuclear magnetic resonance
(NMR) spectroscopy during a prime-constant infusion of
[1-13C]glucose and
[1-13C]mannitol with
either insulin (10 mU · kg
1 · min
1)
or somatostatin (1.0 µg · kg
1 · min
1).
Intracellular glucose was calculated as the difference between the
concentrations of total tissue glucose (calculated from the in vivo
13C NMR spectrum with mannitol as
an internal concentration standard) and extracellular glucose,
corrected by the ratio of intra- and extracellular water space.
Extracellular concentration was corrected for an interstitial
fluid-to-plasma glucose concentration gradient of 0.83 ± 0.07, determined by open-flow microperfusion. The mean ratio of intra- to
extracellular glucose space, determined from the relative NMR signal
intensities and concentrations of mannitol and total creatine, was 9.2 ± 1.1 (hyperglycemic hyperinsulinemia, n = 10), and 9.0 ± 1.7 (hyperglycemic hypoinsulinemia, n = 7). Mean muscle intracellular glucose concentration was <0.07 mM
under hyperglycemic-hyperinsulinemic conditions
(n = 10) and 0.32 ± 0.06 mM under
hyperglycemic-hypoinsulinemic conditions
(n = 7). This method is noninvasive
and should prove useful for resolving the question of whether glucose
transport or phosphorylation is responsible for the reduced rate of
muscle glycogen synthesis observed in diabetic subjects.
nuclear magnetic resonance spectroscopy
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