FASEB J 25: 1088FASEB J 25: -1096FASEB J 25: , 2011. To test if a similar dietary regime would blunt whole body efficiency in endurance-trained men and, as a consequence, hinder aerobic exercise performance, 16 endurance-trained men were given a short-term, high-fat (70% kcal from fat) and a moderate carbohydrate (50% kcal from carbohydrate) diet, in random order. Efficiency was assessed during a standardized exercise task on a cycle ergometer, with aerobic performance assessed during a 1-h time trial and mitochondrial function later measured using 31 P-magnetic resonance spectroscopy. The subjects then underwent a 2-wk wash-out period, before the study was repeated with the diets crossed over. Muscle biopsies, for mitochondrial protein analysis, were taken at the start of the study and on the 5th day of each diet. Plasma fatty acids were 60% higher on the high-fat diet compared with moderate carbohydrate diet (P Ͻ 0.05). However, there was no change in whole body efficiency and no change in mitochondrial function. Endurance exercise performance was significantly reduced (P Ͻ 0.01), most probably due to glycogen depletion. Neither diet led to changes in citrate synthase, ATP synthase, or mitochondrial uncoupling protein 3. We conclude that prior exercise training blunts the deleterious effect of short-term, high-fat feeding on whole body efficiency.exercise; magnetic resonance; nutrition; mitochondria THE MAXIMUM EXTERNAL WORK rate that can be sustained by a muscle or group of muscles is determined by the rate at which adenosine triphosphate (ATP) can be supplied to the myofibrils, with the overwhelming majority of ATP used during sustained exercise being derived from oxidative phosphorylation at the mitochondria. The maximum sustainable rate of mitochondrial ATP supply is, in turn, dependent on the rate of delivery of oxygen and reduced intermediates to the respiratory chain and the mitochondrial phosphorylation-to-oxidation ratio, which describes the economy of energy transduction.The major metabolic fuel sources used by the muscle cell to generate reduced intermediates, and thus ATP, are glucose and fatty acids, with substrate preference being determined by different circumstances. For example, glucose oxidation is stoichiometrically more oxygen efficient than fat oxidation (16), and glycogen breakdown is able to supply ATP at a faster rate than other substrates (32), making it the best fuel when a high sustainable power is required or when oxygen supply is limited. However, energy yield per mole of glucose is low compared with fatty acids, as is the body's total storage capacity for glycogen and glucose (32); thus, in situations in which energetic demands are submaximal but prolonged, and when oxygen is plentiful, fatty acids are the preferred choice.Moreover, free fatty acids (FFA) acutely increase proton leak in isolated mitochondria (33), dissipating the proton gradient as heat, and decreasing the phosphorylation-to-oxidation ratio to a greater extent than that expected from a substrate switch alone. ...