OBJECTIVE: Low-flow ischemia results in glucose transporter translocation and in increased glucose uptake. After total ischemia in rat heart, we found no increase in glucose uptake. Here we test the hypothesis that total ischemia is associated with decreased activation of GLUT4 despite translocation. METHODS: Isolated working hearts (n=70, Sprague-Dawley rats) were perfused for 70 min at physiological workload with Krebs-Henseleit buffer containing [2-3H]glucose (5 mmol/l, 0.05 microCi/ml) with either oleate (0.4 mmol/l, 1%BSA) or pyruvate (5 mmol/l, 1%BSA). After 20 min, hearts were subjected to 15 min of total ischemia followed by 35 min of reperfusion. We measured glucose uptake and intracellular free glucose (IFG) using [2-3H]glucose and [14C]sucrose, and determined the distribution of GLUT4 by colocalization immunofluorescence with Na-K ATP-ase. RESULTS: Cardiac power was 10.1 +/- 0.90 mW before ischemia and did not differ between groups. Recovery was the same in both groups (55.7 +/- 24.8%). Glucose uptake did not differ between groups before ischemia, and did not increase during reperfusion. Despite evidence of GLUT4 translocation after reperfusion in both groups, IFG did not increase compared with before ischemia. CONCLUSION: We conclude that there is a discrepancy between glucose transporter availability and glucose uptake after ischemia, which may be due to inhibition of GLUT4 in the plasma membrane.
OBJECTIVE: Low-flow ischemia results in glucose transporter translocation and in increased glucose uptake. After total ischemia in rat heart, we found no increase in glucose uptake. Here we test the hypothesis that total ischemia is associated with decreased activation of GLUT4 despite translocation. METHODS: Isolated working hearts (n=70, Sprague-Dawley rats) were perfused for 70 min at physiological workload with Krebs-Henseleit buffer containing [2-3H]glucose (5 mmol/l, 0.05 microCi/ml) with either oleate (0.4 mmol/l, 1%BSA) or pyruvate (5 mmol/l, 1%BSA). After 20 min, hearts were subjected to 15 min of total ischemia followed by 35 min of reperfusion. We measured glucose uptake and intracellular free glucose (IFG) using [2-3H]glucose and [14C]sucrose, and determined the distribution of GLUT4 by colocalization immunofluorescence with Na-K ATP-ase. RESULTS: Cardiac power was 10.1 +/- 0.90 mW before ischemia and did not differ between groups. Recovery was the same in both groups (55.7 +/- 24.8%). Glucose uptake did not differ between groups before ischemia, and did not increase during reperfusion. Despite evidence of GLUT4 translocation after reperfusion in both groups, IFG did not increase compared with before ischemia. CONCLUSION: We conclude that there is a discrepancy between glucose transporter availability and glucose uptake after ischemia, which may be due to inhibition of GLUT4 in the plasma membrane.
Authors: Johannes S Kern; Stefan Loeckermann; Anja Fritsch; Ingrid Hausser; Wera Roth; Thomas M Magin; Claudia Mack; Marcel L Müller; Oliver Paul; Patrick Ruther; Leena Bruckner-Tuderman Journal: Mol Ther Date: 2009-06-30 Impact factor: 11.454
Authors: Anja Fritsch; Stefan Loeckermann; Johannes S Kern; Attila Braun; Michael R Bösl; Thorsten A Bley; Hauke Schumann; Dominik von Elverfeldt; Dominik Paul; Miriam Erlacher; Dirk Berens von Rautenfeld; Ingrid Hausser; Reinhard Fässler; Leena Bruckner-Tuderman Journal: J Clin Invest Date: 2008-05 Impact factor: 14.808