Haseler, Luke J., Michael C. Hogan, and Russell S. and in chronic disease conditions such as cardiac
Richardson. Skeletal muscle phosphocreatine recovery in failure that are known to result in reduced mitochon-
exercise-trained humans is dependent on O2 availability. J. drial content and oxidative capacity (3840). In con-
Appl. Physiol. 86(6): 20132018, 1999.In skeletal muscle, trast, PCr recovery was enhanced with endurance
phosphocreatine (PCr) recovery from submaximal exercise training in athletes (22, 26, 42), consistent with the
has become a reliable and accepted measure of muscle increased mitochondrial content and activities of the
oxidative capacity. During exercise, O2 availability plays a
enzymes associated with oxidative metabolism allow-
role in determining maximal oxidative metabolism, but the
relationship between O2 availability and oxidative metabo-
ing a greater capacity for oxidative generation of ATP
lism measured by 31P-magnetic resonance spectroscopy (MRS) (6, 14, 24). Additionally, McCully et al. (27) have
during recovery from exercise has never been studied. We demonstrated a linear relationship between PCr recov-
used 31P-MRS to study exercising human gastrocnemius ery and citrate synthase activity in human skeletal
muscle under conditions of varied fractions of inspired O2 muscle. More recently, Paganini et al. (29) demon-
(FIO2) to test the hypothesis that varied O2 availability strated in rats that the rate constant for PCr recovery
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2014 PHOSPHOCREATINE RECOVERY WITH VARIED FIO2
continuously, allowing the calculation of arterial O2 satura- Fourier transformation. All spectra were manually phased by
tion (assuming no alveolar to arterial PO2 gradient and no using zero- and first-order phase corrections. There were no
significant metabolic acidosis). Heart rate was monitored phase variations among rest, exercise, and recovery during
continuously throughout the experiment with a finger probe the experiment. The levels of PCr determined from the
(Omni-Trak, In Vivo Research). In each FIO2 (0.1, 0.21, and intensity of that peak were normalized to 100% by using the
1.00), subjects performed a 5-min warm-up period followed by average value obtained from the last 40 s of rest acquired for
5 min of rest, and then they performed 5 min of exercise each subject as a reference. Muscle intracellular pH was
followed by 5 min of recovery. Subjects were allowed 40 min of calculated from the chemical shift difference () of the Pi peak
rest between each complete exercise bout. The order of the relative to the PCr peak by using the following equation:
three treatments was varied to allow all six possible orders to pH 6.75 log[( 3.27)/(5.69 )] (37). Signal-to-noise
be performed once and to minimize any ordering effects. ratios (20:1) were sufficient to allow PCr levels to be
Throughout the study subjects were unaware of the treat- determined with a temporal resolution of 4 s during exercise
ment order. and recovery. Changes in PCr during recovery were fit to a
31P-MRS. MRS was performed by using a clinical 1.5-T monoexponential function
General Electric Signa system (version 4.8) operating at
25.86 MHz for 31P. 31P-MRS data were acquired with a PCr(t) PCr0 PCr1 [1 e(tTD/)]
transmit/receive surface coil (diameters 20 and 10 cm, respec-
tively) placed under the calf at its maximum diameter. The where PCr0 is the baseline value, PCr1 is the difference
centering of the leg over the coil was confirmed by T1- between the baseline and the recovery value, t is time, TD is
weighted 1H localizing images obtained in the axial plane. the time delay, and is the time constant.
Magnetic field homogeneity was optimized by shimming on Statistical analysis. Data were tested with repeated-
the proton signal from tissue water. For 31P-MRS the pulse measures ANOVA (Tukey post hoc) by using a commercially
ample of limited mitochondrial function due to O2 linear dependence of 1/ on oxidative capacity (27, 29).
supply (32, 35). It is noteworthy that both of these Because [PCrrest] is constant for a given subject, the
theories are complementary and may be somewhat rate constant for PCr recovery, Qmax, and maximal O2
interelated. The latter theory is supported by previous consumption (VO2 max) are all indicative of the maximal
studies that have observed slowed PCr recovery due to rate of oxidative ATP synthesis. Thus it is not surpris-
a reduction in the arterial PO2 (1) and in cases of ing that there is a strong similarity between O2 con-
reduced muscle blood flow (9) (discussed in detail in sumption and PCr on- and off-exercise kinetics (25) or
Evidence of mitochondrial O2 supply-dependent ATP that both Qmax and VO2 max are linearly dependent on
synthesis). However, the former theory is supported by muscle oxidative capacity (15, 29). Consequently, PCr
the observation that altered tissue oxygenation during exercise-recovery data are clearly indicative of both
steady-state submaximal exercise results in altered muscle VO2 max and muscle oxidative capacity: a greater
PCr levels, suggesting that tissue or intracellular oxy- oxidative capacity leads to a greater capacity to con-
genation plays a role in modulating regulators of sume O2 and a shorter PCr and vice versa. The
cellular respiration (10, 11). Thus cellular levels of O2 present data provide the first MRS evidence of a
may influence metabolic control even during submaxi- dissociation between the first two variables (muscle
mal exercise. It is also known that exercise training VO2 max and oxidative capacity) due to altered FIO2. This
increases the mitochondrial content of the cell (12, 13), is evident by the fact that the rate constant for PCr
and this alters the mitochondrial sensitivity to the recovery (similar in many respects to VO2 max) is altered
regulators of respiration (7). Thus mitochondrial con- by varying O2 availability while oxidative capacity
tent itself has been shown to play a role in controlling remained unchanged due to the acute nature of the
the PCr rate constant on oxygenation seen in Fig. 3 into the cell suggests that under normal conditions the
suggests that PCr recovery from submaximal exercise recovery of PCr is limited by O2 supply. This can be
is limited by O2 availability. The observation here that interpreted as further evidence that diffusion of O2
PCr recovery is enhanced with increased intracellular from erythrocyte to mitochondria, and ultimately intra-
oxygenation provides evidence that under normoxic cellular PO2, plays an important role in determining
conditions Qmax is determined by O2 availability and not skeletal muscle VO2 max. Finally, the practical implica-
mitochondrial metabolic limits. Figure 3 provides fur- tion of these data is that PCr recovery measurements
ther evidence of an in vivo correlate of the effect of O2 should be interpreted with caution because differences
tension on cellular respiration rate (32), as originally in between subjects may not be due to metabolic
demonstrated in vitro by Wilson et al. (41). These data limitations but rather to variations in O2 availability.
suggest that the dependence of the rate constant for Hence a lengthened exhibited in a diseased state may
PCr recovery on O2 availability may be approaching a be due to O2 supply limitations and not to a metabolic
plateau whereby further increments in intracellular abnormality.
PO2 will have a diminishing effect. These findings are
The authors thank the subjects for their time in volunteering for
consistent with the concept that VO2 max is dependent on this study and Kuldeep Tagore for valuable technical assistance.
the availability of O2 (31, 35). This research was supported by National Institutes of Health
There is prior evidence in untrained subjects perform- Grants HL-17731 and AR-40155. R. S. Richardson was a Parker B.
ing small-muscle-mass exercise that suggests the effect Francis Fellow in Pulmonary Research during this study.
Address for reprint requests and other correspondence: L. J.
of varying FIO2 on O2 delivery is dampened by an Haseler, Dept. of Medicine 0623A, Univ. of California, San Diego,
alteration in muscle blood flow (36). In that case, 9500 Gilman Dr., La Jolla, CA 92093-0623 (E-mail: lhaseler
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