Mourtzakis, Marina, Bengt Saltin, Terry Graham, and Henriette Pilegaard. Carbohydrate metabolism during prolonged exercise and recovery: interactions between pyruvate dehydrogenase, fatty acids, and amino acids. J Appl Physiol 100: 1822-1830, 2006. First published January 19, 2006 doi:10.1152/japplphysiol.00571.2005.-During prolonged exercise, carbohydrate oxidation may result from decreased pyruvate production and increased fatty acid supply and ultimately lead to reduced pyruvate dehydrogenase (PDH) activity. Pyruvate also interacts with the amino acids alanine, glutamine, and glutamate, whereby the decline in pyruvate production could affect tricarboxycylic acid cycle flux as well as gluconeogenesis. To enhance our understanding of these interactions, we studied the time course of changes in substrate utilization in six men who cycled at 44 Ϯ 1% peak oxygen consumption (mean Ϯ SE) until exhaustion (exhaustion at 3 h 23 min Ϯ 11 min). Femoral arterial and venous blood, blood flow measurements, and muscle samples were obtained hourly during exercise and recovery (3 h). Carbohydrate oxidation peaked at 30 min of exercise and subsequently decreased for the remainder of the exercise bout (P Ͻ 0.05). PDH activity peaked at 2 h of exercise, whereas pyruvate production peaked at 1 h of exercise and was reduced (ϳ30%) thereafter, suggesting that pyruvate availability primarily accounted for reduced carbohydrate oxidation. Increased free fatty acid uptake (P Ͻ 0.05) was also associated with decreasing PDH activity (P Ͻ 0.05) and increased PDH kinase 4 mRNA (P Ͻ 0.05) during exercise and recovery. At 1 h of exercise, pyruvate production was greatest and was closely linked to glutamate, which was the predominant amino acid taken up during exercise and recovery. Alanine and glutamine were also associated with pyruvate metabolism, and they comprised ϳ68% of total amino-acid release during exercise and recovery. Thus reduced pyruvate production was primarily associated with reduced carbohydrate oxidation, whereas the greatest production of pyruvate was related to glutamate, glutamine, and alanine metabolism in early exercise. alanine; pyruvate dehydrogenase kinase 4 mMRNA; carbohydrate oxidation; nitrogen balance THE REGULATION OF CARBOHYDRATE provision during prolonged exercise and recovery is a profound challenge for the human body. Muscle carbohydrate metabolism does not happen in isolation but requires integration across tissues as well as regulation with the other major substrates, such as fats and amino acids. Because carbohydrate stores are limited in the body (i.e., muscle, liver), the highly coordinated integration of liver, adipose tissue, pancreas, and muscle metabolism is important for constant energy provision during prolonged exercise (1, 30, 34 -36) and in reestablishing homeostasis during recovery (2). Arterial glucose delivery is one key element in energy provision, and the body attempts to maintain glucose delivery during exercise by activating liver glucose production with decreased insulin, increased gl...