J Appl Physiol 103: 177–183, 2007.
First published April 12, 2007; doi:10.1152/japplphysiol.01460.2006.
Effects of acute hypoxia on cerebral and muscle oxygenation during
incremental exercise
Andrew W. Subudhi, Andrew C. Dimmen, and Robert C. Roach
University of Colorado Altitude Research Center, Denver Health Science Center and Colorado Springs Campuses, Colorado
Submitted 29 December 2006; accepted in final form 11 April 2007
Subudhi AW, Dimmen AC, Roach RC. Effects of acute hypoxia
on cerebral and muscle oxygenation during incremental exercise.
J Appl Physiol 103: 177–183, 2007. First published April 12, 2007;
doi:10.1152/japplphysiol.01460.2006.—To determine if fatigue at
maximal aerobic power output was associated with a critical decrease
in cerebral oxygenation, 13 male cyclists performed incremental
maximal exercise tests (25 W/min ramp) under normoxic (Norm: 21%
FIO2) and acute hypoxic (Hypox: 12% FIO2) conditions. Near-infrared
spectroscopy (NIRS) was used to monitor concentration (␮M)
changes of oxy- and deoxyhemoglobin (⌬[O2Hb], ⌬[HHb]) in the
left vastus lateralis muscle and frontal cerebral cortex. Changes in
total Hb were calculated (⌬[THb] ⫽ ⌬[O2Hb] ⫹ ⌬[HHb]) and used
as an index of change in regional blood volume. Repeated-measures
ANOVA were performed across treatments and work rates (␣ ⫽
0.05). During Norm, cerebral oxygenation rose between 25 and 75%
peak power output {Powerpeak; increased (inc) ⌬[O2Hb], inc. ⌬[HHb],
inc. ⌬[THb]}, but fell from 75 to 100% Powerpeak {decreased (dec)
⌬[O2Hb], inc. ⌬[HHb], no change ⌬[THb]}. In contrast, during
Hypox, cerebral oxygenation dropped progressively across all work
rates (dec. ⌬[O2Hb], inc. ⌬[HHb]), whereas ⌬[THb] again rose up to
75% Powerpeak and remained constant thereafter. Changes in cerebral
oxygenation during Hypox were larger than Norm. In muscle, oxy-
genation decreased progressively throughout exercise in both Norm
and Hypox (dec. ⌬[O2Hb], inc. ⌬ [HHb], inc. ⌬[THb]), although
⌬[O2Hb] was unchanged between 75 and 100% Powerpeak. Changes
in muscle oxygenation were also greater in Hypox compared with
Norm. On the basis of these findings, it is unlikely that changes in
cerebral oxygenation limit incremental exercise performance in nor-
moxia, yet it is possible that such changes play a more pivotal role in
hypoxia.
near-infrared spectroscopy; fatigue; altitude
RECENT ATTENTION has been drawn to the hypothesis that a
critical reduction in cerebral oxygenation may pose a central
limitation to exercise performance by inhibiting cortical acti-
vation of efferent motoneurons (1, 11, 25, 28, 42, 44). Invasive
methods used to assess cerebral oxygenation have presented
obstacles to testing this hypothesis directly; however, near-
infrared spectroscopy (NIRS) offers noninvasive, real-time
measurement of tissue oxygenation (27) to overcome these
difficulties.
Light in the near-infrared spectrum readily penetrates skin,
fat, and bone and is differentially absorbed by heme-containing
molecules in underlying tissues, predominantly oxy- and de-
oxyhemoglobin (O2Hb, HHb; Ref. 53). A modified form of the
Beer-Lambert law quantifies changes in O2Hb and HHb con-
centrations over time to reflect the balance between regional
oxygen delivery and extraction in the illuminated area.
Address for reprint requests and other correspondence: A. W. Subudhi,
Dept. of Biology, Univ. of Colorado at Colorado Springs, 1420 Austin Bluffs
Parkway, Colorado Springs, CO 80918 (e-mail: asubudhi@uccs.edu).
http://www. jap.org 8750-7587/07 $8.00 Copyright © 2007 the American Physiological Society
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NIRS measurements have been well correlated with intra-
cellular oxygen tension in muscle (50) and venous oxygen
saturation in both muscle (18, 53) and cerebral tissue (22).
Several authors have reported that muscle oxygenation de-
creases progressively with increased exercise intensity, reach-
ing a minimum or plateau near maximal aerobic power output
(3–7, 21, 35, 38, 53). Changes in cerebral oxygenation have not
been as well defined. In general, cerebral oxygenation has been
shown to increase during submaximal exercise, but either
remains elevated (24, 25, 32, 39, 41) or decreases at maximal
intensity (19, 26, 40). While Gonzalez-Alonso et al. (19)
argued that reductions in cerebral oxygenation do not represent
a limit to performance, Nielsen et al. (40) reported that admin-
istration of supplemental oxygen (FIO2 ⫽ 30%) maintains ce-
rebral oxygenation and increases time trial performance with-
out affecting muscle oxygenation, thus supporting a central
limitation to exercise. Saito et al. (46) and Imray et al. (25)
suggested that cerebral hypoxia may be a limiting factor to
exercise under certain environmental conditions, as their sub-
jects exhibited reductions in cerebral oxygenation while per-
forming exercise at high altitudes. More recently, Rasmussen
et al. (44) showed that decreased frontal cortex oxygenation
was associated with reduced muscle force-generating capacity.
Although these studies demonstrated an association between
cerebral oxygenation and exercise performance, no studies
have determined if a critical level of cerebral deoxygenation
limits performance.
A standard model that clearly differentiates between central
(central nervous system) and peripheral (muscle) factors asso-
ciated with fatigue during intense, aerobic exercise has yet to
be established. The intent of this study was to use NIRS to
monitor changes in both central and peripheral oxygenation
during incremental exercise to exhaustion. The study was the
first to monitor simultaneous changes in cerebral and muscle
oxygenation under both normoxic and hypoxic conditions. We
specifically tested the hypothesis that a critical change in
cerebral, but not muscle, oxygenation would immediately pre-
cede voluntary cessation of exercise.
MATERIALS AND METHODS
Subjects
Following institutional review board approval, competitive cyclists
were recruited through local cycling clubs. Respondents were
screened to select active competitors with recent experience cycling at
elevations between 2,500 and 4,300 m. Fifteen male cyclists, meeting
the above criteria, gave their written informed consent to participate in
the study. Prior to the experimental protocol, all subjects were given
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177
178 CEREBRAL AND MUSCLE OXYGENATION DURING EXERCISE
a complete physical examination, including blood tests (complete
blood count and metabolic panel), to assess general health. Two
subjects were excluded from the study based on the results of baseline
examinations (hypertension and recent pneumonia). Thirteen cyclists
completed the experimental protocol.
Experimental Design
The study was conducted using a single blinded, cross-over design.
Subjects first performed a practice trial of incremental exercise to
maximal exertion under ambient conditions (22°C; 15% humidity,
625 mmHg). The testing protocol was repeated on two additional
days, spaced at least 1 wk apart, while breathing either normoxic
(Norm; FIO2 ⫽ 21%; PIO2 ⬃ 131 mmHg) or hypoxic (Hypox; FIO2 ⫽
12%; PIO2 ⬃ 75 mmHg) mixtures of dry medical grade gas. The order
of treatment was randomly assigned and counterbalanced.
Exercise Testing
All testing was performed on a Velotron cycle ergometer (Racer-
mate, Seattle, WA), which was adjusted to each cyclist’s specifica-
tions. Prior to testing, subjects were required to warm up for 20 min,
under ambient conditions, at a work rate (watts) equaling 1.5 ⫻ body
weight (kg). Subjects were then instrumented with necessary probes,
sensors, and breathing apparatus. To prime the breathing system and
analyzers, subjects inhaled experimental gas for no more than 2 min
prior to the commencement of exercise. Initial work rate was set at 25
W and increased at a ramped rate of 25 W/min until subjects reached
perceived exhaustion, as indicated by volitional cessation of exercise,
or failure to maintain a pedal cadence of ⬎50 rpm despite strong
verbal encouragement. Following the test, all instrumentation was
removed and subjects performed self-paced cool down under ambient
conditions.
Instrumentation
NIRS theory. Determination of tissue oxygenation via NIRS is
based on the Beer-Lambert Law, which quantifies the attenuation of
light traveling through a non-scattering medium. It is mathematically
expressed as OD␭ ⫽ log(Io/I) ⫽ ⑀␭ 䡠 c 䡠 Lo, where OD␭ is the medium’s
optical density, Io is the incident light intensity, I is the transmitted
light intensity, ⑀␭ is the extinction coefficient of the chromophore
(mM⫺1 䡠 cm⫺1), c is the concentration of the chromophore (mM⫺1),
Lo is the distance (cm) between light entry and exit, and ␭ is the
wavelength of light used.
When applied to biological tissues, the equation is modified to account
for light scattering (16): OD␭ ⫽ log(Io/I) ⫽ ⑀␭ 䡠c䡠Lo 䡠DPF ⫹ OD␭x.
The differential path length factor (DPF) is given by L/Lo, where L
is the average distance traveled by each photon. OD␭x accounts for
attenuation due to scattering and absorption by other chromophores. If
OD␭x is assumed to be constant, one can solve for change in concen-
tration using the formula, ⌬c ⫽ ⌬OD␭/(⑀␭ 䡠 Lo 䡠 DPF). This equation is
valid for a scattering medium with one chromophore. Additional
wavelengths of light must be used to differentiate between chro-
mophores in more complex systems.
In biological tissues, NIR light is specifically absorbed by heme
groups within hemoglobin (Hb) and myoglobin (Mb); thus NIRS
cannot differentiate between the two species. Since Hb is tetrameric
and exists in appreciably greater concentrations than the monomeric
Mb species, the relative contribution of Hb is substantially greater
than that of Mb (53). The ability of NIRS to quantify changes in
oxygenation is based on the difference in absorption spectra for
oxygenated and deoxygenated heme groups. In light of these facts, it
is appropriate to refer to NIRS measurements as indexes of overall
tissue oxygenation (51), with the understanding that underlying vas-
cular beds are composed of ⬃70% venous blood (8).
NIRS measurements. During all exercise sessions, subjects were
instrumented with two pairs of NIR probes to monitor absorption of
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light across cerebral and muscle tissue (Oxymon, Artinis, The Neth-
erlands). Headsets were constructed to hold one NIR emitter and
detector pair over the left frontal cortex region of the forehead.
Spacing between optodes was adjusted to ensure proper placement
and signal strength on each individual (range 4.26 – 4.61 cm). A
second emitter and detector pair was affixed over the belly of the left
vastus lateralis muscle (⬃15 cm above the proximal border of the
patella and 5 cm lateral to the midline of the thigh). Skinfold
measurements were made in the sagittal plane midway between
optodes to account for skin and adipose thickness. Probes were held
in place by a plastic spacer with fixed optode distance of 5.18 cm and
secured to the skin using double-sided tape. Elastic bandages were
used to shield the optodes from ambient light. The Beer-Lambert law
was used to calculate micromolar changes in tissue oxygenation
(⌬[O2Hb] and ⌬[HHb]) across time using received optical densities
from two continuous wavelengths of NIR light (780 and 850 nm) and
published DPFs of 4.95 (17) and 5.93 (52) for muscle and cerebral
tissue, respectively. Total Hb (⌬[THb]) was calculated as the sum of
⌬[O2Hb] and ⌬[HHb] and used as an index of change in regional
blood volume (51). Data were recorded at 10 Hz and filtered with a
Savitzky-Golay smoothing algorithm prior to analysis. Both cerebral
and muscle measurements were normalized to reflect changes from
the beginning of the exercise protocol (arbitrarily defined as 0 ␮M) to
express the magnitude of deoxygenation near maximal exercise. A
postexercise, leg cuff-ischemia technique to obtain a low-oxygenation
reference point was not used because Chance et al. (12) reported little
additional deoxygenation (⬍2%) during surpasystolic cuff occlusion
of the vastus lateralis muscle immediately following incremental
exercise to maximal exertion.
Absolute measurements of hemoglobin concentration and percent
saturation values (analogous to those obtained via pulse oximetry)
were unattainable because the exact DPFs were unknown. Thus direct
comparisons between the two tissues could not be made due to
differences in tissue composition and volumes illuminated.
By definition, [O2Hb] and [HHb] exist in equilibrium, such that an
increase in one results in a stoichiometric decrease in the other. Thus
we interpret decreases in ⌬[O2Hb] and increases in ⌬[HHb] as
evidence of relative deoxygenation and, vice versa, as evidence of
improved oxygenation. These explanations are only valid if regional
blood volume, given by ⌬[THb], is constant. Since arterial blood is
mostly oxygenated, changes in regional arterial flow are reflected by
parallel changes in ⌬[O2Hb] if the rate of oxygen consumption
remains constant. Because ⌬[HHb] is closely associated with changes
in venous oxygen content and is less sensitive to ⌬[THb] than
⌬[O2Hb] is, ⌬[HHb] is believed to be a sensitive measure of relative
tissue deoygenation due to oxygen extraction (20).
Cardiopulmonary measurements. Heart rate was measured and
recorded with a chest strap and monitor (Polar USA, Irvine, CA).
Subjects were instructed to keep their hands and fingers relaxed
during exercise testing to obtain strong, pulsatile, finger-tip oximetry
signals from either the left index or middle finger (Criticare 503,
Waukesha, WI). Experimental gases (Norm ⫽ compressed air;
Hypox ⫽ 12% O2, 88% N2) were administered using a system of
plastic tubing and 200 liter Douglas bag reservoir. Subjects’ expired
breath was directed through 1.8 m of tubing to a 3.0 liter mixing
chamber. Ventilation was measured using a heated pneumotach (Hans
Rudolph series 4813, Kansas City, MO). Gas samples from the mixing
chamber were analyzed with O2 (Ametek S-3A, AEI Technologies,
Pittsburgh, PA) and CO2 (Ametek CD-3A, AEI Technologies) ana-
lyzers. Analog data from each instrument were continuously recorded
with a PowerLab 16 data-acquisition system (ADIntruments, Colo-
rado Springs, CO), sampling at 100 Hz. Metabolic variables of interest
(V̇O2, V̇CO2) were calculated offline using the Haldane transformation
to obtain average values over 20-s periods. Ventilatory threshold (VT)
was determined using ventilatory equivalents of O2 and CO2, as
previously described (2). Peak power output (Powerpeak) was defined
as power output recorded immediately before termination of exercise.
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 CEREBRAL AND MUSCLE OXYGENATION DURING EXERCISE
Analyses
Metabolic responses to exercise during Norm and Hypox were
evaluated with paired t-tests. All NIRS data were expressed as
micromolar concentration change from rest, immediately before the
start of exercise (0 ␮M). Mean values (⌬[O2Hb], ⌬[HHb], and
⌬[THb]) were plotted against relative power output to qualitatively
describe cerebral and muscle oxygenation patterns during exercise for
each treatment. Reliability of NIRS data in response to exercise under
normoxic conditions (practice test and Norm) was assessed using the
Intraclass Correlation Coefficient (ICC) Alpha. Pearson product mo-
ment correlations were used to evaluate relationships between NIRS
and pulse oximeter measurements.
Quantitative differences in cerebral and muscle NIRS measure-
ments were analyzed with multivariate (Wilk’s Lambda), repeated-
measures ANOVA to evaluate effects of treatment (Norm and Hypox)
across both relative work rates (25%, 50%, 75%, and 100% Peak
Powerpeak) and absolute work rates (100 and 200 W). Criterion for
significance was set at P ⬍ 0.05. Post hoc, pairwise comparisons were
made using the Holm’s sequential method to control for type I error.
RESULTS
Subject Characteristics
Subjects were competitive, amateur cyclists who resided at
altitudes between 1,585 and 1,890 m and reported weekly
training volumes of 11 ⫾ 3 h during the testing period
(January-April). Those completing all experimental trials
(n ⫽ 13) were 30 ⫾ 7 yr old, 182 ⫾ 6 cm tall, weighed 78 ⫾
9 kg, and had thigh skinfold thickness of 8 ⫾ 3 mm.
Average Hb concentration and hematocrit were 16.2 ⫾ 0.7
g/dl and 47 ⫾ 2%.
Metabolic Measurements
Average metabolic responses to exercise trials are shown in
Table 1. V̇O2peak (ml䡠kg⫺1 䡠min⫺1) and Powerpeak (W) were
reduced 37 and 29% under Hypox compared with Norm (P ⬍
0.05). Hypox resulted in relative hyperventilation at absolute
work rates, but ventilation at the point of exhaustion was lower
than Norm (Table 1). V̇O2 and Power at VT were 33% lower
during Hypox (P ⬍ 0.05), but, in relative terms, were not
different from Norm (70 ⫾ 5% V̇O2peak and 63 ⫾ 5%
Powerpeak during Norm vs. 74 ⫾ 8% V̇O2peak and 59 ⫾ 7%
Powerpeak during Hypox). All but two subjects were able to
correctly identify the order of treatment; however, no subjects
reported adverse side effects of hypoxia (dizziness, headache,
or nausea) during or following exercise.
Table 1. Metabolic responses to incremental exercise tests
Normoxia Hypoxia
Variable VT Peak VT
V̇E, l/min 85.8⫾10.3 179.7⫾27.1 84.9⫾9.0 152.2⫾19.3*
V̇O2, ml 䡠 kg⫺1 䡠 min⫺1 42.6⫾5.0 60.7⫾5.6 28.4⫾4.8* 38.1⫾4.0*
V̇E/V̇O2 25.6⫾2.4 39.7⫾3.5 37.8⫾3.2* 54.6⫾5.0*
V̇E/V̇CO2 27.1⫾2.3 35.0⫾3.5 38.0⫾3.5* 46.6⫾4.1*
Power, W 256⫾30 409⫾39 171⫾16* 291⫾26*
SpO2, % 92⫾3 91⫾3 77⫾12* 73⫾11*
HR, beats/min 150⫾13 179⫾10 146⫾12 166⫾10*
Values are means ⫾ SD; n ⫽ 13. VT, ventilatory threshold. *Different from
normoxia (P ⬍ 0.05).
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179
Near-Infrared Measurements
Reliability of NIRS measurements was evaluated using data
collected during practice and Norm trials. ICC Alpha was high
for cerebral (0.98 to 0.99) and muscle (0.97 to 0.98) ⌬[O2Hb],
⌬[HHb], and ⌬[THb] across all work rates. Cerebral and
muscle NIRS measurements were correlated (P ⬍ 0.001) with
pulse oximeter readings during exercise in Norm (r ⫽ 0.62 to
0.90) and Hypox (r ⫽ 0.94 to 0.96). Average concentration
changes in cerebral and muscle NIRS signals were graphed
for all subjects across relative work rates during Norm and
Hypox (Fig. 1).
Cerebral analysis. During Norm, regional cerebral oxygen-
ation was unchanged from rest to 25% Powerpeak. NIRS signals
increased from 25 to 75% Powerpeak (1⌬[O2Hb],1⌬[HHb],
and 1⌬[THb]), suggesting regional cerebral vasodilation
during moderate intensity exercise, yet displayed evi-
dence of decreased oxygenation between 75 and 100%
Powerpeak while regional blood volume remained constant
(2⌬[O2Hb],1⌬[HHb], 7⌬[THb]; Table 2). During Hypox,
regional cerebral oxygenation decreased progressively from
the start of exercise to the point of maximal exertion
(2⌬O2Hb,1⌬HHb), whereas regional blood volume again
increased, but reached a plateau after 75% Powerpeak. Changes
in ⌬[O2Hb] and ⌬[HHb] were greater in Hypox compared with
Norm at all relative and absolute work rates (Tables 2 and 3).
Between Norm and Hypox, there was no change in ⌬[THb]
with respect to absolute work rate (Table 3), but ⌬[THb] was
lower in Hypox at work rates ⬎75% Powerpeak (Table 2).
Muscle analysis. During Norm, regional muscle oxygen-
ation decreased progressively from the start of exercise to 75%
Powerpeak (2⌬[O2Hb],1⌬[HHb]), despite increased regional
blood volume (1⌬[THb]). From 75 to 100% Powerpeak,
⌬[O2Hb] was unchanged, yet ⌬[HHb] and ⌬[THb] rose
slightly, indicating a reduction in the rate of muscle deoxygen-
ation during high-intensity exercise. All NIRS variables
reached plateaus at work rates ⬎90% Powerpeak (Fig. 1). In
Hypox, regional muscle oxygenation followed a similar pattern
as seen in Norm; however, ⌬[HHb] and ⌬[THb] did not reach
plateaus near maximal exercise intensity. The extent of muscle
deoxygenation (2⌬[O2Hb],1⌬[HHb]) was greater than Norm
at all work rates, whereas changes in regional blood volume
were not different between Norm and Hypox.
DISCUSSION
The major finding in this study was that hypoxia profoundly
affected the pattern of frontal cerebral cortex oxygenation
during incremental exercise. Results demonstrated that incre-
mental exercise to maximal exertion elicited a larger degree of
cerebral deoxygenation in hypoxia compared with normoxia.
This study was the first to monitor simultaneous changes in
cerebral and muscle oxygenation during incremental exercise
Peak
to maximal exertion under both normoxic and acute hypoxic
conditions. We tested the hypothesis that a critical change in
cerebral oxygenation would immediately precede voluntary
cessation of exercise. Under normoxic conditions, we detected
a drop in cerebral, but not muscle, oxygenation during late-
stage, high-intensity exercise; however, during acute hypoxia,
a similar decrease in cerebral oxygenation was observed during
low-intensity exercise, long before the point of exhaustion.
Thus we conclude that it is unlikely that frontal cortex oxy-
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180 CEREBRAL AND MUSCLE OXYGENATION DURING EXERCISE

Fig. 1. Near-infrared spectroscopy (NIRS) concentration

changes (means ⫾ SE) during incremental exercise in
normoxia (F) and hypoxia (E). All values are expressed
relative to the peak power output achieved in normoxia.

genation limits exercise performance in normoxia, but ac-
knowledge the notion that it may play a more pivotal role in
hypoxia (1, 25, 44).
Technical Considerations
Influence of skin blood flow. Because NIRS probes are
typically placed on the skin surface, the relative contribution of
skin blood flow to NIRS tissue signals has been questioned (10,
14). The influence of skin blood flow has been estimated to
account for ⬃15 to 23% of NIR light attenuation (31, 49), thus
underlying tissues are the primary determinants of resulting
NIRS measurements. Owen-Reece et al. (43) and Kirkpatrick
et al. (29) demonstrated that occlusion of scalp blood flow does
not affect underlying cerebral NIRS measurements when op-
tode distance is sufficient. Since NIR light follows an arc-
shaped path through tissues and penetrates to a depth of
approximately one-half of the distance between emitter and
detector, wider optode spacing decreases the relative contri-
bution from skin. We used an optode distance comparable to
that of Owen-Reece et al. (43) to minimize the effect of skin
blood flow. Nevertheless, if our ⌬[O2Hb] and ⌬[THb] mea-
surements were elevated as a result of augmented skin blood
flow during exercise, our results would underestimate the
relative extent of tissue deoxygenation and not alter our
conclusions.
Reliability of NIRS measurements. While oxygenation is not
homogeneous across skeletal muscle beds (36), we minimized
potential errors introduced by probe placement/replacement by
measuring and tracing the final location of the probes with an
indelible marker. The cerebral NIRS probe was placed ipsilat-
erally on the forehead, just lateral to the midline. The location
and distance between emitter and detector was adjusted for
each individual to obtain strong, pulsatile NIRS signals on first
placement. Handmade headsets, which held NIRS probes
firmly, were customized for each individual to avoid move-
ment artifact during exercise and minimize errors in sequential
placements. NIRS measurements were limited to the frontal
cortex, yet frontal lobe oxygenation has been associated with
global changes in cerebral blood flow (9), metabolic rate (37),
and muscle force (44) during hypoxia.

Table 2. NIRS concentration changes across relative work rates

Normoxia Hypoxia

Variable 25% 50% 75% 100% 25% 50% 75% 100%

Cerebral concentrations, ␮M
⌬关O2Hb兴 ⫺1.6⫾0.8 1.6⫾1.5† 8.2⫾2.1†‡ 1.8⫾3.1§ ⫺6.96⫾1.3* ⫺13.8⫾1.7*† ⫺16.1⫾1.7*†‡ ⫺19.7⫾1.8*†‡§
⌬关HHb兴 1.0⫾0.3 1.3⫾0.6 5.3⫾1.1†‡ 13.6⫾1.2†‡§ 9.1⫾0.9* 18.7⫾1.2*‡ 24.0⫾1.7*†‡ 27.6⫾2.0*†‡§
⌬关THb兴 ⫺0.6⫾0.8 2.9⫾1.2† 13.4⫾1.8†‡ 15.4⫾2.9†‡ 2.1⫾1.0* 4.9⫾1.4† 7.9⫾1.9*†‡ 7.9⫾2.2*†‡
Muscle concentrations, ␮M
⌬关O2Hb兴 ⫺10.8⫾2.3 ⫺18.4⫾2.8† ⫺30.0⫾3.3†‡ ⫺30.4⫾4.2†‡ ⫺18.5⫾1.4* ⫺35.7⫾3.4*† ⫺46.0⫾3.7*†‡ ⫺46.6⫾3.6*†‡
⌬关HHb兴 9.9⫾2.7 23.8⫾2.8† 39.3⫾3.7†‡ 48.1⫾5.0†‡§ 16.0⫾2.8* 41.6⫾4.7*† 55.2⫾5.2*†‡ 66.3⫾6.0*†‡§
⌬关THb兴 ⫺0.9⫾1.9 5.4⫾1.6† 9.3⫾1.6†‡ 17.6⫾2.3†‡§ ⫺2.4⫾2.7 5.9⫾2.5† 9.2⫾2.6†‡ 19.7⫾4.0†‡§
Values are micromolar changes from resting conditions (means ⫾ SE); n ⫽ 13. NIRS, near-infrared spectrometry; O2Hb, oxyhemoglobin; HHb,
deoxyhemoglobin; THb, total hemoglobin. Brackets indicate concentration. Different from *normoxia, †25%, ‡50%, §75% (P ⬍ 0.05).

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 CEREBRAL AND MUSCLE OXYGENATION DURING EXERCISE 181
Table 3. NIRS concentration changes across work rate during acute hypoxia and would also immediately
absolute work rates precede the termination of exercise.
The decrease in cerebral oxygenation from the start of
Normoxia Hypoxia
exercise to 25% Powerpeak in hypoxia was similar to that seen
Variable 100 W 200 W 100 W 200 W near maximal exercise intensity in normoxia. If our hypotheses
were correct, subjects would have stopped exercising in hyp-
Cerebral concentrations, ␮M
oxia at only ⬃25% of their actual Powerpeak. In reality, sub-
⌬[O2Hb] ⫺0.9⫾0.8 1.3⫾1.7 ⫺11.1⫾1.4* ⫺16.2⫾1.6*†
⌬[HHb] 0.7⫾0.3 1.7⫾0.6 13.8⫾1.1* 23.3⫾1.5*† jects continued to perform at progressively higher work rates in
⌬[THb] ⫺0.2⫾0.7 2.9⫾1.4 2.7⫾1.0 7.1⫾1.7† the face of continual reductions in cerebral oxygenation. The
Muscle concentrations, ␮M extent of deoxygenation (2⌬[O2Hb],1⌬[HHb]) was, there-
⌬[O2Hb] ⫺12.8⫾2.2 ⫺20.2⫾2.5† ⫺25.8⫾2.2* ⫺45.4⫾3.8*† fore, greater in hypoxia throughout exercise, despite minimal
⌬[HHb] 11.5⫾2.3 25.7⫾2.9† 27.3⫾2.8* 54.4⫾5.2*† changes in regional blood volume (⌬[THb]) at comparable
⌬[THb] ⫺1.3⫾1.9 5.5⫾1.7† 1.5⫾2.2 9.0⫾2.4† work rates.
Values are micromolar changes from resting conditions (means ⫾ SE); These findings demonstrate a large tolerance for change in
n ⫽ 13. Different from *normoxia, †100 W (P ⬍ 0.01). cerebral oxygenation during exercise and refute the thought
that smaller changes seen during normoxia limited perfor-
mance. Our results do not discount the possibility that cerebral
Our test retest validation of NIRS changes during normoxic oxygenation is an important variable in the integrative decision
exercise trials shows a high degree of reliability for changes in to stop exercise (34) nor can we refute the notion that such
tissue oxygenation across all work rates (ICC range: 0.97– large changes in cerebral oxygenation may limit exercise
0.99). These findings are in agreement with those of Kishi et al. performance in hypoxia (1, 25, 44). Further tests with more
(30) and Kolb et al. (33), who reported 8 –9.4% day-to-day severe hypoxia or that rapidly alter cerebral oxygenation but
variation in NIRS measurements of cerebral oxygenation. maintain systemic normoxia are necessary to this hypothesis.
Additionally, investigations with chronic exposures to hypoxia
Cerebral Oxygenation During Incremental Exercise are necessary to determine the effect of acclimatization on
We observed that regional cerebral oxygenation and blood cerebral oxygenation during exercise.
volume were maintained during low-intensity exercise up to
25% Powerpeak and increased between 25 and 75% Powerpeak Muscle Oxygenation During Incremental Exercise
(1⌬[O2Hb],1⌬[HHb], and 1⌬[THb]). These findings are in
Vastus lateralis oxygenation decreased linearly up to
agreement with the theory that cerebral perfusion increases to
75% Powerpeak, despite increased regional blood volume
maintain oxygen delivery during submaximal exercise (24 –26,
32, 39). At higher exercise intensities (75 to 100% Powerpeak), (2⌬[O2Hb],1⌬[HHb],1⌬[THb]). At higher exercise intensi-
cerebral oxygenation fell in all subjects. Specifically, from 75 ties, the rate of deoxygenation decreased as all values reached
to 100% Powerpeak, a progressive decrease in ⌬[O2Hb] and plateaus above 90% Powerpeak. While some authors have
reciprocal increase in ⌬[HHb] was observed while ⌬[THb] reported that a rightward shift (Bohr effect) in the oxyhemo-
remained unchanged. These results are in accord with others globin dissociation curve accelerates the rate of deoxygenation
who reported decreased cerebral oxygenation during maximal at or near lactate or ventilatory thresholds (3, 21, 36), our
(40) and supramaximal (47) exercise protocols. Such high results appeared more linear at these exercise intensities. NIRS
exercise intensities are associated with hyperventilation-in- measurements have been correlated with changes in intracel-
duced reductions in arterial CO2 tension, which results in lular oxygen tension (50) and venous oxygen saturation (18,
cerebral vasoconstriction and diminished cerebral blood flow 53), thus plateaus in oxygenation have been interpreted as
(25, 40). Gonzalez-Alonso et al. (19) suggested that the fall in evidence of maximal oxygen extraction (18, 48, 53), in accor-
cerebral oxygenation near exhaustion during constant work dance with the theory that diffusion rate of oxygen across
rate tests is due to a small decrease in cerebral blood flow myocyte membranes is limited at exercise intensities ⬎50%
paired with a relatively larger increase in cerebral O2 uptake. Powerpeak (45). Since NIRS cannot distinguish between hemo-
Our data support a similar conclusion for incremental exercise globin and myoglobin, we cannot make specific claims about
tests. Near maximal exercise intensity, the rate of change in the rate of oxygen transfer between blood and muscle. We
regional cerebral blood volume was reduced (from 1⌬[THb] contend that plateaus in oxygenation observed in the present
to 7⌬[THb]), whereas the rate of oxygen extraction was study simply imply a balance between overall oxygen de-
accelerated (2⌬[O2Hb] and 1⌬[HHb]). livery and extraction/consumption over progressively higher
work rates. Because subjects were able to continue exercis-
Does Cerebral Oxygenation Limit Incremental ing at greater intensities after the appearance of the plateau,
Exercise Performance? it is unlikely that observed levels of tissue deoxygenation
We questioned whether the observed drop in cerebral oxy- were directly responsible for the termination of exercise
genation near maximal exercise intensity in normoxia triggered during normoxic conditions. Our assessment is supported by
the cessation of exercise. To test this hypothesis, we compared the findings of Nielsen et al. (40), who showed that supple-
the patterns of change in cerebral oxygenation obtained during mental oxygen did not affect the pattern of muscle deoxy-
normoxia and acute hypoxia. We hypothesized that a similar genation during exercise despite a significant improvement
drop in cerebral oxygenation would be observed at a lower in performance.
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182 CEREBRAL AND MUSCLE OXYGENATION DURING EXERCISE
Effect of Acute Hypoxia on Muscle Oxygenation
During hypoxia, patterns of change in muscle oxygenation
were similar to normoxia up to 75% Powerpeak, but differed
thereafter. Further inspection of individual data revealed that
only 6 of the 13 subjects demonstrated plateaus in oxygenation
near maximal exercise intensity. Cyclists who demonstrated
plateaus tended to achieve significantly greater maximal power
output during normoxia (P ⫽ 0.06), but displayed larger
reductions in maximal power output between normoxia and
hypoxia (P ⫽ 0.04) than the athletes who continued to deox-
ygenate throughout exercise. We detected no other physiolog-
ical differences between groups, but note that these post hoc
observations were limited by low statistical power. Thus future
studies are warranted to explain individual differences in mus-
cle oxygenation patterns during acute hypoxia and test their
association with fatigue.
The extent of tissue deoxygenation during hypoxia was
greater at all relative and absolute work rates. These results are
similar to those who report larger levels of deoxygenation
during constant work rate tests under hypoxic conditions (13,
23, 35) and add support to our claim that tissues maintain their
functional ability under a wide range of oxygenation. Our
findings are in contrast to those who report minimal changes in
tissue oxygenation between normoxic and hypoxic trials at a
given absolute work rate (15), potentially highlighting differ-
ences in patterns of oxygenation between incremental and
constant work rate tests.
In conclusion, the results of this NIRS study demonstrate
that incremental exercise performance under normoxic condi-
tions is not likely to be limited by changes in cerebral oxygen-
ation, at least in athletic residents of moderate altitude. How-
ever, these findings do not discount potential moderating ef-
fects that tissue oxygenation may play in a complex,
integrative model of fatigue. Future studies should investigate
the effects of varied and more severe levels of hypoxia with
innovative methods that independently alter cerebral and mus-
cle oxygenation during exercise to gain further insight into the
role that tissue oxygenation may play in fatigue.
ACKNOWLEDGMENTS
We express our gratitude to Paige Sheen for her assistance in project
management and both Ben Honigman and Vaughn Browne for their medical
expertise during subject recruitment and screening.
GRANTS
This project was funded in part by National Heart, Lung, and Blood
Institute Grant HL-070362.
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