Publications (3)3.3 Total impact
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ABSTRACT: The effects of deprivation and supplementation of exogenous glutamine (0.06 and 2.2 mM in the culture medium, respectively) were studied in mononucleated myoblasts and in multinucleated myotubes. Myoblasts cultured in glutamine-deprived medium showed reductions in plating efficiency and myotube fusion index. Myotubes grown in glutamine-supplemented cultures had higher intracellular glutamine concentrations than those grown in glutamine-deprived medium (67 +/- 4.2 vs. 46 +/- 3.6 nmol/mg cell protein, respectively) and glutamine-supplemented myotubes utilized glutamine, whereas glutamine-deprived myotubes released it. Glutamine deprivation for 12 h caused a significant, cycloheximide-blockable increase in the capacity for glutamine uptake via system Nm in both myoblasts and myotubes (maximum velocity increases of 23 +/- 5.3 and 35 +/- 4.2%, respectively), which was reversed by glutamine replenishment. Depriving myotubes of glutamine did not alter the kinetics of uptake of amino acid transport systems A, ASC, or L. Glutamine deprivation resulted in a threefold increase in glutamine synthetase activity, whereas glutaminase activity remained unchanged. System Nm and glutamine synthetase appear to undergo adaptive upregulation in glutamine-deprived muscle cells to compensate for the reduced exogenous glutamine supply.The American journal of physiology 01/1994; 265(6 Pt 1):E935-42.
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ABSTRACT: Glutamine transport was studied in preconfluent monolayered, mononucleated myoblasts (4 days old) and in fused, multinucleated, differentiated myotubes (10 days old), both prepared from neonatal rat skeletal muscle. The initial (60 s) rate of 50 microM glutamine uptake in myoblasts and myotubes was stereospecific, saturable, and largely (80%) Na+ dependent. At glutamine concentrations of 0.01-1 mM, Na(+)-dependent uptake showed saturation kinetics: in myoblasts, the Michaelis constant (Km) was 197 +/- 38 microM, maximum velocity (Vmax) was 1,165 +/- 60 pmol.min-1.mg protein-1; in myotubes, Km was 174 +/- 51 microM and Vmax was 1,435 +/- 47 pmol.min-1.mg protein-1. The Na(+)-dependent glutamine uptake was Li+ tolerant in both myoblasts and myotubes. The Na(+)-dependent uptake of 50 microM L-[3H]glutamine was investigated in the presence of various amino acids at 0.01-10 mM. Histidine and asparagine competitively inhibited glutamine uptake, but inhibition by serine was noncompetitive; glutamate, arginine, leucine, and 2-aminobicyclo(2,2,1)heptane-2-carboxylate (BCH) had no significant inhibitory effects; 2-(methyl-amino)isobutyrate (MeAIB) caused a small but significant inhibition. In parallel with a stimulation of glucose transport, addition of insulin stimulated Na(+)-dependent glutamine uptake within 1 h by a maximum of 27% in myoblasts and 42% in myotubes (half-maximal stimulation at 0.3 nM insulin). Glucagon had no effect. Kinetic analysis revealed that the insulin-stimulated increase in glutamine transport was due to a Vmax effect, which was cycloheximide inhibitable. The insulin-stimulated increase was Li+ tolerant and not inhibited by MeAIB or cysteine at 1 mM. The results indicate that the predominant glutamine transporter of neonatal rat skeletal muscle cells in primary tissue culture in System Nm. System Nm also appears to be the major insulin-sensitive glutamine transport component in skeletal muscle. Primary muscle culture appears to be a useful preparation for studying glutamine transport and its regulation.The American journal of physiology 08/1993; 265(1 Pt 1):E135-44.
- Clinical Nutrition 03/1993; 12(1):53-4. · 3.30 Impact Factor