Metabolic transformation of rabbit skeletal muscle cells in primary culture in response to low glucose.

We have investigated the mechanism of the changes in the profile of metabolic enzyme expression that occur in association with fast-to-slow transformation of rabbit skeletal muscle. The hypotheses assessed are: do 1) lowered intracellular ATP concentration or 2) reduction of the muscular glycogen stores act as triggers of metabolic transformation? We find that 3 days of decreased cytosolic ATP content have no impact on the investigated metabolic markers, whereas incubation of the cells with little or no glucose leads to decreases in glycogen in conjunction with decreases in glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter activity, GAPDH mRNA and specific GAPDH enzyme activity (indicators of the anaerobic glycolytic pathway), and furthermore to increases in mitochondrial acetoacetyl-CoA thiolase (MAT, also known as ACAT) promoter activity, peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) expression and citrate synthase (CS) specific enzyme activity (all indicators of oxidative metabolic pathways). The AMP-activated protein kinase (AMPK) activity under these conditions is reduced compared to controls. In experiments with two inhibitors of glycogen degradation we show that the observed metabolic transformation caused by low glucose takes place even if intracellular glycogen content is high. These findings for the first time provide evidence that metabolic adaptation of skeletal muscle cells from rabbit in primary culture can be induced not only by elevation of intracellular calcium concentration or by a rise of AMPK activity, but also by reduction of glucose supply. Contrary to expectations, neither an increase in phospho-AMPK nor a reduction of muscular glycogen content are crucial events in the glucose-dependent induction of metabolic transformation in the muscle cell culture system studied.