Fuel use during glycogenesis in rainbow trout (Oncorhynchus mykiss Walbaum) white muscle studied in vitro

Abstract

The purpose of this study was to examine fuel used during muscle glycogenesis in rainbow trout Oncorhynchus mykiss using an in vitro muscle slice preparation to test the hypothesis that intracellular lactate is the major glycogenic substrate and the muscle relies upon extracellular substrates for oxidation. Fish were exhaustively exercised to reduce muscle glycogen content, muscle slices were taken from exhausted fish and incubated for 1 h in medium containing various substrates at physiological concentrations. 14C-labeled lactate, glycerol or palmitate was added and 14C incorporation into muscle glycogen and/or CO2 was measured. Lactate clearance in the absence of net glycogenesis suggests that when suitable oxidizable extracellular substrates were lacking, intracellular lactate was oxidized. Only muscle incubated in lactate, glycerol or palmitate synthesized glycogen, with the greatest synthesis in muscle incubated in lactate plus glycerol. The major fate of these extracellular substrates was oxidative, with lactate oxidized at rates 10 times that of palmitate and 100 times that of glycerol. Neither extracellular lactate nor glycerol contributed significantly to glycogenesis, with lactate carbon contributing less than 0.1% of the total glycogen synthesized, and glycerol less than 0.01%. There was 100 times more extracellular lactate-carbon incorporated into CO2 than into glycogen. In the presence of extracellular lactate, palmitate or glycerol, intracellular lactate was spared an oxidative fate, allowing it to serve as the primary substrate for in situ glycogenesis, with oxidation of extracellular substrates driving ATP synthesis. The primary fate of extracellular lactate is clearly oxidative, while that of intracellular, glycolytically derived lactate is glycogenic, which suggests intracellular compartmentation of lactate metabolism.