Intracellular oxygen (O 2 ) availability and the impact of ambient hypoxia have far reaching ramifications in terms of cell signalling and homeostasis; however, in vivo cellular oxygenation has been an elusive variable to assess. Within skeletal muscle the extent to which myoglobin desaturates (deoxy-Mb) and the extent of this desaturation in relation to O 2 availability provide an endogenous probe for intracellular O 2 partial pressure (P iO 2 ). By combining proton nuclear magnetic resonance spectroscopy ( 1 H NMRS) at a high field strength (4 T), assessing a large muscle volume in a highly efficient coil, and extended signal averaging (30 min) we assessed the level of skeletal muscle deoxy-Mb in 10 healthy men (30 ± 4 years) at rest in both normoxia and hypoxia (10% O 2 ). In normoxia there was an average deoxy-Mb signal of 9 ± 1%, which, when converted to P iO 2 using an O 2 /Mb half-saturation (P 50 ) of 3.2 mmHg, revealed an P iO 2 of 34 ± 6 mmHg. In ambient hypoxia the deoxy-Mb signal rose to 13 ± 3% (P iO 2 = 23 ± 6 mmHg). However, intersubject variation in the defence of arterial oxygenation (S aO 2 ) in hypoxia (S aO 2 range: 86-67%) revealed a significant relationship between the changes in S aO 2 and P iO 2 (r 2 = 0.5). These data are the first to document resting intracellular oxygenation in human skeletal muscle, highlighting the relatively high P iO 2 values that contrast markedly with those previously recorded during exercise (∼2-5 mmHg). Additionally, the impact of ambient hypoxia on P iO 2 and the relationship between changes in S aO 2 and P iO 2 stress the importance of the O 2 cascade from air to cell that ultimately effects O 2 availability and O 2 sensing at the cellular level.

T 1 -based determination of perfusion was performed with the high temporal and spatial resolution that monitoring of exercise physiology requires. As no data were available on the validation of this approach in human muscles, T 1 -based NMRI of perfusion was compared to standard strain-gauge venous occlusion plethysmography performed simultaneously within a 4 T magnet. Two different situations were investigated in 21 healthy young volunteers: 1) a 5-min ischemia of the leg, or 2) a 2-3 min ischemic exercise consisting of a plantar flexion on an amagnetic ergometer. Leg perfusion was monitored over 5-15 min of the recovery phase, after the air-cuff arterial occlusion had been released. The interesting features of the sequence were the use of a saturation-recovery module for the introduction of a T 1 modulation and of single-shot spin echo for imaging. Spatial resolution was 1.7 ؋ 2.0 mm and temporal resolution was 2 s. For data analysis, ROIs were traced on different muscles and perfusion was calculated from the differences in muscle signal intensity in successive images. To allow comparison with the global measurement of perfusion by plethysmography, the T 1 -based NMR measurements in exercising muscles were rescaled to the leg cross-section.