Validity and accuracy of calculating oxidative ATP synthesis in vivo during high-intensity skeletal muscle contractions

Abstract

Several methods have been developed for using ³¹ P-MRS to calculate rates of oxidative ATP synthesis (ATP_OX ) during muscular contractions based on assumptions that (1) the ATP cost of force generation (ATP_COST ) remains constant or (2) Michaelis-Menten coupling between cytosolic ADP and ATP_OX does not change. However, growing evidence suggests that one, or both, of these assumptions are invalid during high-intensity fatigue protocols. Consequently, there is a need to examine the validity and accuracy of traditional ATP_OX calculation methods under these conditions. To address this gap, we measured phosphate concentrations and pH in the vastus lateralis muscle of nine young adults during four rest-contraction-recovery trials lasting 24, 60, 120, and 240 s. The initial velocity of phosphocreatine resynthesis (V_iPCr ) following each trial served as the criterion measure of ATP_OX because this method makes no assumptions of constant ATP_COST or Michaelis-Menten coupling between changes in cytosolic ADP and ATP_OX . Subsequently, we calculated ATP_OX throughout the 240 s trial using several traditional calculation methods and compared estimations of ATP_OX from each method with time-matched measurements of V_iPCr . Method 1, which assumes that ATP_COST does not change, was able to model changes in V_iPCr over time, but showed poor accuracy for predicting V_iPCr across a wide range of ATP_OX values. In contrast, Michaelis-Menten methods, which assume that the relationship between changes in cytosolic ADP and ATP_OX remains constant, were invalid because they could not model the decline in V_iPCr . However, adjusting these Michaelis-Menten methods for observed changes in maximal ATP_OX capacity (i.e., V_max ) permitted modeling of the decline in V_iPCr and markedly improved accuracy. The results of these comprehensive analyses demonstrate that valid, accurate measurements of ATP_OX can be obtained during high-intensity contractions by adjusting Michaelis-Menten ATP_OX calculations for changes in V_max observed from baseline to post-fatigue.

Keywords: 31P-MRS, ATP cost, bioenergetics, high-intensity exercise, mitochondria, muscle, oxidative capacity, oxidative phosphorylation.