Baroreex responsiveness during ventilatory acclimatization in humans

Brian E. Hunt,
1
Renaud Tamisier,
2,3
Geoffrey S. Gilmartin,
1
Mathew Curley,
1
Amit Anand,
1
and J. Woodrow Weiss
1
1
Department of Kinesiology, School of Public Health and Health Sciences, University of Massachusetts, Amherst,
Massachusetts;
2
Pulmonary and Sleep Research Laboratory, Division of Pulmonary, Critical Care and Sleep Medicine,
Beth Israel Deaconess Medical Center, Boston, Massachusetts; and
3
Laboratoire du Sommeil, Laboratoire HP2 (Hypoxie
PathoPhysiologie), Centre Hospitalier Universitaire de Grenoble, Grenoble, France
Submitted 7 February 2008; accepted in nal form 25 August 2008
Hunt BE, Tamisier R, Gilmartin GS, Curley M, Anand A, Weiss
JW. Baroreex responsiveness during ventilatory acclimatization in
humans. Am J Physiol Heart Circ Physiol 295: H1794H1801, 2008.
First published August 29, 2008; doi:10.1152/ajpheart.131.2008.We
tested the hypothesis that the decline in muscle sympathetic activity
during and after 8 h of poikilocapnic hypoxia (Hx) was associated with
a greater sympathetic baroreex-mediated responsiveness. In 10 healthy
men and women (n 2), we measured beat-to-beat blood pressure
(Portapres), carotid artery distension (ultrasonography), heart period, and
muscle sympathetic nerve activity (SNA; microneurography) during two
baroreex perturbations using the modied Oxford technique before,
during, and after 8 h of hypoxia (84% arterial oxygen saturation). The
integrated baroreex response [change of SNA (SNA)/change of dia-
stolic blood pressure (DBP)], mechanical (diastolic diameter/DBP),
and neural (SNA/diastolic diameter) components were esti-
mated at each time point. Sympathetic baroreex responsiveness
declined throughout the hypoxic exposure and further declined
upon return to normoxia [pre-Hx, 8.3 1.2; 1-h Hx, 7.2 1.0;
7-h Hx, 4.9 1.0; and post-Hx: 4.1 0.9 arbitrary integrated
units (AIU) min
1
mmHg
1
; P 0.05 vs. previous time point for
1-h, 7-h, and post-Hx values]. This blunting of baroreex-mediated
efferent outow was not due to a change in the mechanical
transduction of arterial pressure into barosensory stretch. Rather,
the neural component declined in a similar pattern to that of the
integrated reex response (pre-Hx, 2.70 0.53; 1-h Hx,
2.59 0.53; 7-h Hx, 1.60 0.34; and post-Hx, 1.34 0.27
AIU min
1
m
1
; P 0.05 vs. pre-Hx for 7-h and post-Hx values).
Thus it does not appear as if enhanced baroreex function is primarily
responsible for the reduced muscle SNA observed during intermediate
duration hypoxia. However, the central transduction of baroreceptor
afferent neural activity into efferent neural activity appears to be reduced
during the initial stages of peripheral chemoreceptor acclimatization.
autonomic
AUTONOMIC RESPONSES TO HYPOXIA vary greatly depending on the
intensity, duration, and pattern of the stimulus (31). Transient
exposure to hypoxia, as little as 20 min, results in an increase
in minute ventilation that returns to prehypoxia levels within
several minutes after exposure, whereas vascular sympathetic
activity remains elevated for at least an hour after return to
normoxia (26, 48). Prolonged exposure to hypoxia (weeks to
months) results in an elevated minute ventilation and muscle
sympathetic activity, and both are sustained for at least several
days after return to normoxia (2, 6, 15). Although much of the
work regarding autonomic regulation in humans has focused
on the effects of either transient or prolonged hypoxia, little is
known about the transitional period between short-term and
prolonged hypoxia. The few data available suggest that when
the hypoxic exposure is extended to several hours, minute
ventilation demonstrates a further increase that persists after a
return to normoxia, whereas sympathetic activity returns to
preexposure levels (10, 31).
Transient hypoxia elicits a rapid increase in pulmonary
ventilation via direct increases in afferent input from peripheral
chemoreceptors. This is followed by a modest decline or roll
off in ventilation over the course of 2030 min, likely caused
by reduced sensitivity of arterial chemoreceptors to PO
2
(1, 34).
Although the mechanisms underlying the rapid and sustained
sympathoexcitation associated with transient hypoxia are not
well understood, the baroreex regulation of vascular sympa-
thetic activity appears well maintained and not a likely candi-
date (12). Prolonged hypoxia, several hours to days, results in
a time-dependent increase in minute ventilation, termed venti-
latory acclimatization to hypoxia (VAH), which outlasts the
exposure. Previous data suggest that VAH begins by 4 h of
hypoxic exposure (43). Increased sensitivity of both the arterial
chemoreceptor and the central nervous system to PO
2
are
thought to underlie this chronic response (31). However, the
mechanisms underlying the return of sympathetic activity to
prehypoxic levels during the initial stages of VAH are unclear.
As chemoreceptors are sympathoexcitatory, the dissociation
between augmented peripheral chemoreceptor sensitivity and
reduced sympathetic neural outow during VAH argues
against a direct chemoreceptor-mediated response. However,
given the close proximity of carotid chemoreceptor and sym-
pathoinhibitory baroreceptor afferent projections into the me-
dulla, it is plausible that the increased medullary activity
associated with VAH may augment baroreex restraint of
muscle sympathetic activity during this period. Therefore, we
tested the hypothesis that sympathetic baroreex-mediated
responsiveness would be augmented during and after 8 h of
poikilocapnic hypoxia.
METHODS
Study Sample
Twelve healthy, nonsmoking, normotensive volunteers, free of vaso-
active medications, completed the study (women 3). Adequate sym-
pathetic neurograms were obtained in 10 volunteers. All study volunteers
underwent a routine medical history and physical examination to exclude
overt cardiopulmonary or neurological disease before giving written
informed consent. To eliminate possible confounding hormonal effects
on cardiovascular function, the women were studied during the week
Address for reprint requests and other correspondence: B. E. Hunt, Applied
Health Science, Wheaton College, 501 College Ave., Wheaton, IL 60187
(e-mail: brian.hunt@wheaton.edu).
The costs of publication of this article were defrayed in part by the payment
of page charges. The article must therefore be hereby marked advertisement
in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Am J Physiol Heart Circ Physiol 295: H1794H1801, 2008.
First published August 29, 2008; doi:10.1152/ajpheart.131.2008.
0363-6135/08 $8.00 Copyright 2008 the American Physiological Society http://www.ajpheart.org H1794

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following menses and all tested negative for pregnancy via human
chorionic gonadotropin test. This protocol conformed to the provisions of
the Declaration of Helsinki and was thus approved by the Institutional
Review Board at the Beth Israel Deaconess Medical Center.
Experimental Protocol
All studies were conducted between 9:00 AM and 7:00 PM after study
volunteers had fasted overnight. After instrumentation, the volunteers
rested in the supine position for 1530 min until all parameters were
stable for at least 5 min. During the following 45 min, continuous
baseline recordings were made of sympathetic activity, arterial pressure,
and electrocardiogram (ECG) while the subjects breathed room air. In
addition, two consecutive baroreex perturbations using the modied
Oxford technique were performed (8). We then exposed the volunteers to
continuous poikilocapnic hypoxia (Hx). The volunteers breathed through
a leak-free face mask (8940 Series; Hans Rudolph, Kansas City, MO)
tted with a Hans Rudolph 2600 series two-way valve. Athree-way valve
on the inspiratory tubing allows rapid switching between the 30-liter
respiratory bag and room air. The respiratory bag was lled with a gas
mixture containing 9% oxygen balanced with nitrogen. Nitrogen or room
air was added to the inspired gas as necessary to maintain arterial oxygen
saturation (Sa
O
2
) near 85%. During the exposure, the measurements of
blood pressure, ECG, and two baroreex perturbations were performed
after 1 and 7 h. During the hypoxic exposure study, the volunteers were
allowed to drink water ad libitum and were continuously infused with 5%
glucose at a rate of 100 ml/h. After 8 h, the volunteers were returned to
room-air breathing and all measurements were repeated after Sa
O
2
re-
turned to baseline, with baroreex testing beginning at 30 min of
posthypoxia.
Measurements
Cardiovascular and pulmonary variables. Heart rate was derived
from the ECG. Clinical arterial pressure was measured in the right arm
using an automated arm-cuff sphygmomanometer (Dinamap model,
Critikon, Tampa, FL). Continuous beat-by-beat blood pressure measure-
ments by photoplethysmography (Portapres TNO-Institute of Applied
Physics Biomedical Instrumentation, Amsterdam, The Netherlands) were
used in the calculation of baroreex responsiveness. Sa
O
2
was monitored
with a pulse oximeter (Biox model 3740; Ohmeda, Louisville, CO), and
CO
2
fraction was measured continuously using an infrared gas analyzer
connected to the mask (model 17630; Vacumed, Ventura, CA). End-tidal
CO
2
(ET
CO
2
) was calculated as the product of the Fe CO
2
value at the end
of the expiratory plateau and barometric pressure. Data from previous
subjects exposed to the same 8 h of poikilocapnic hypoxia (10) demon-
strated that ET
CO
2
, fraction of inspired oxygen (FI
O
2
), and Sa
O
2
were not
different between the 1- and 7-h time points, suggesting the Sa
O
2
is an
adequate index of the hypoxic stress in our healthy study volunteers. The
change in ET
CO
2
across time points within an individual was used as a
gross index of peripheral chemoreex drive.
Muscle sympathetic nerve activity. We obtained nerve recordings
using standard tungsten microelectrodes inserted into the peroneal
nerve posterior in the popliteal area after localization by surface
stimulation. The signals were ltered, amplied, and full-wave recti-
ed. The rectied signal was integrated for display on an oscilloscope
and for recording (Nerve Trafc Analyzer, Model 662c-3, Bioengi-
neering Dept., University of Iowa, Iowa City, IA). The electrode
position in the muscle bers was conrmed by pulse synchronous
bursts of activity occurring 1.21.4 s after the QRS complex, repro-
ducible activation during the second phase of the Valsalva maneuver,
elicitation of afferent nerve activity by mild muscle stretching, and the
absence of response to startle. We recorded the sympathetic neuro-
gram before hypoxia and continued the recordings for 1 h after
hypoxia was begun. The electrodes were then removed. The micro-
electrodes were replaced, and the sympathetic neurogram was re-
corded after 7 h of hypoxia and continued recording for 1 h after
volunteers returned to breathing room air. Sympathetic bursts were
identied by the Hamner-Taylor detection algorithm (14) using Mat-
lab software (Mathworks, Natick, MA). Burst amplitude was normal-
ized by assigning a value of 1,000 arbitrary units to the largest burst
identied during stable baseline activity before each measurement;
this was expressed as both frequency (in bursts/min) and rate (in
bursts/100 heart beats). Both metrics are stable and reproducible
indexes and thus are used to compare sympathetic activity between
time points. The total burst activity was then estimated by calculating
the area under each sympathetic burst; this was expressed as arbitrary
integrated units (AIU) and used to express the change in sympathetic
activity during baroreex perturbations. Since the Hamner-Taylor
algorithm has not been independently validated, we compared the
sympathetic burst frequency as determined by the semiautomated
Hamner-Taylor algorithm to manual burst detection in 14 neurograms
(7 steady-state normoxia and 7 steady-state hypoxia). The burst
frequency determined by both methods was highly correlated (r
2

0.79) with the automated detection having a xed bias of 1.5

bursts/min.
Baroreex engagement. We used the modied Oxford technique,
which involves a bolus injection of 100 g of sodium nitroprusside
followed in 60 s by a bolus of 150 g of phenylephrine hydrochloride.
This technique generally produces an initial 15 mmHg fall in
arterial pressure followed by an 15 mmHg rise in pressure above
resting supine levels (8, 17). Responses of a representative study
volunteer are shown in Fig. 1. During baroreex engagement, the
beat-by-beat arterial pressures from a nger photoplethysmograph
(Portapres), a standard three-lead ECG, and muscle sympathetic nerve
activity (MSNA) were recorded continuously at a sampling rate of
500 Hz. Longitudinal B-mode images of a common carotid artery just
below the carotid bulb were also obtained during baroreex engage-
ment using a 7.5-MHz transducer. Commercially available hardware
(3155 PCI Frame Grabber, Data Translations, Marlboro, MA) and
software (DVI Acquisition, Information Integrity, Stow, MA) were
used to acquire 30-Hz images to a computer triggered from the
R-wave of the ECG. Fifteen consecutive carotid images were acquired
Fig. 1. Blood pressure (BP), R-R interval, carotid diameter, and muscle
sympathetic nerve activity (MSNA) from a single study volunteer during
baroreex engagement before hypoxia.
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with each trigger to approximate at least one third of the cardiac cycle
(i.e., 500 ms of a 1,500-ms R-R interval or 40 beats/min heart rate)
and therefore encompass both end-diastolic and peak-systolic diam-
eters. Each 640 480-pixel image provided a resolution of 0.05
mm/pixel. We used custom software that employed a verterbi search
algorithm to estimate carotid diameters. Two trials were performed at
each time point. Each trial was separated by at least 15 min to allow
for the recovery of blood pressure and heart rate to the preinjection
levels.
For each trial, the diameters were associated to the appropriate blood
pressures, R-R intervals, and sympathetic activity (29, 37). The data were
then averaged across 2-mmHg increments or bins. We excluded bins in
the threshold and saturation regions in estimating the linear responsive-
ness of the integrated, mechanical, and neural aspects of the baroreex.
Integrated cardiovagal baroreex responsiveness was estimated from the
slope of the relation between R-R interval and systolic blood pressure
(R-R interval/systolic blood pressure). The mechanical component of
the vagal limb was expressed as the slope of the relation between systolic
blood pressure and carotid systolic diameter (systolic blood pressure/
systolic diameter). The neural component of the vagal limb was ex-
pressed as the slope of the relation between R-R interval and carotid
systolic diameter (R-R interval/systolic diameter). Each aspect of the
sympathetic limb was characterized in a similar fashion: integrated
responsiveness (MSNA/diastolic blood pressure), mechanical (dia-
stolic blood pressure/diastolic diameter), and neural (MSNA/dia-
stolic diameter). Figure 2 depicts how estimates of baroreex responsive-
ness were calculated from the data shown in Fig. 1.
Because central or peripheral hysteresis can affect the slope of
these relations, we independently assessed the slopes during the
falling and rising portions of the response. No systematic differences
were found between slopes during the falling and rising pressures.
Therefore, the slopes were estimated from data encompassing the fall
and rise in blood pressure. Baroreex responsiveness at each time point is
expressed as the mean of the two trials (average trial-to-trial difference of
0.30 ms/mmHg for vagal and 0.20 AIU burst
1
mmHg
1
for sympa-
thetic gains).
Data Analysis
All comparisons were limited to data sets from the 10 volunteers in
which adequate pre- and postsympathetic neurograms were obtained.
A one-way ANOVA for repeated measures was used to compare all
data across the four time points of the study. Newman-Keuls post hoc
analysis was used when ANOVA indicated differences between time
Fig. 2. Vagal baroreex responses (left) and sympathetic
baroreex responses (right) derived from data depicted in Fig.
1. Top: integrated response. Middle: mechanical component.
Bottom: neural component of the integrated baroreex re-
sponse. SBP, systolic BP; DBP, diastolic BP.
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points. Trends were determined by orthogonal polynomials (11, 16).
Data are reported as means SE in the text, table, and gures.
P values 0.05 were considered statistically signicant.
RESULTS
As depicted in Fig. 3, MSNA initially increased before when
VAH has been reported to begin (43), then returned to prehy-
poxia levels at 7 h of exposure, and declining further at
postexposure. Along with sympathetic activity, Table 1 dis-
plays the hypoxic stimulus as measured by Sa
O
2
and the
attendant changes in heart rate, ET
CO
2
, and mean arterial
pressure throughout the study.
There was a linear trend for integrated vagal baroreex
responsiveness to decline across time (P 0.07), with the
lowest value recorded after volunteers were returned to room-
air breathing (P 0.08; Fig. 4). There was no corresponding
systematic difference in either the mechanical component (pre-
Hx, 2.43 2.23; 1-h Hx, 2.45 2.94; 7-h Hx, 1.83 2.43;
and post-Hx, 2.06 1.22 m/mmHg) or neural component
(pre-Hx, 8.85 1.84; 1-h Hx, 7.38 1.58; 7-h Hx, 8.93
1.92; post-Hx, 6.51 1.05 ms/m) of the reex arc.
Integrated sympathetic baroreex responsiveness declined
during the hypoxic exposure, achieving its lowest measured
value at 30 to 45 min posthypoxia (Fig. 4). There was no
difference in the mechanical component across time (pre-Hx,
3.23 0.30; 1-h Hx, 3.28 0.30; 7-h Hx, 2.95 0.20; and
post-Hx, 2.97 0.40 m/mmHg), as depicted in Fig. 5.
However, hypoxia was associated with a decline in the neural
component, with a further decline observed after returning to
Table 1. Cardiovascular and ventilatory variables
Prehypoxia 1-h Hypoxia 7-h Hypoxia Posthypoxia
Arterial O2 Saturation, % 98.80.8 83.93.8* 83.34.2* 98.50.8
End-tidal CO2, mmHg 44.72.7 40.34.2* 40.93.6* 39.53.2*
Heart Rate, beats/min 5813 7419* 7818* 6211
MSNA, bursts/100 beats 2610 3210* 218* 166*
MAP, mmHg 868 878 8610 918
Values are means SE. MSNA, muscle sympathetic nerve activity; MAP,
mean arterial pressure measured in the brachial artery. *P 0.05 vs. prehyp-
oxia.
Fig. 3. MSNA and BP responses before and
during baroreex from a single study volunteer
at each time point. Steady-state MSNA (left)
and integrated sympathetic baroreex response
(SBR; right) are shown. AU, arbitrary unit.
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room-air breathing (pre-Hx, 2.70 0.53; 1-h Hx, 2.59
0.53; 7-h Hx, 1.60 0.34; and post-Hx, 1.34 0.27
AIU beat
1
m
1
; P 0.05 vs. pre-Hx for 7-h and post-Hx
values).
Carotid diastolic diameter increased during the rst hour of
hypoxia (pre-Hx, 6.00 0.15 vs. 6.32 0.23 mm, P 0.05)
and continued to be greater after 7 h of hypoxia (6.34 0.22
mm). Upon the resumption of room-air breathing, the diameter
was not different from prehypoxia (5.93 0.17 mm).
Although there was no relation between sympathetic barore-
ex responsiveness and sympathetic activity before or after just
1 h of hypoxia, the greater sympathetic baroreex responsive-
ness was associated with lower sympathetic bursting rate
across individuals at 7 h of hypoxia (r 0.89, P 0.05) and
after returning to room-air breathing (r 0.79, P 0.05).
However, within individuals, these variables were not consis-
tently related across time points, with correlation coefcients
ranging from 0.92 to 0.99 (mean 0.23 0.32). Further-
more, the baroreex responsiveness was not related to periph-
eral chemoreex drive, as suggested by ET
CO
2
, with intra-
individual correlation coefcients ranging from 0.26 to 0.57,
with all P values 0.23.
DISCUSSION
We hypothesized that the decline in sympathetic activity
seen after 8 h of poikilocapnic hypoxia would be associated
with increased sympathetic baroreex responsiveness. How-
ever, our data fail to support this hypothesis. Rather, they
indicate that baroreex responsiveness declined during hyp-
oxia and was at its lowest after returning to room-air breathing.
This reduction does not appear to be related to changes in the
mechanical transduction of arterial pressure into the barorecep-
tor stretch within the carotid artery. Rather, altered neural
properties within the reex arc may underlie this unexpected
reduction in responsiveness. We are aware of no previous data
providing insight into changes in the baroreex function during
short-term VAH.
Most data regarding the effects of hypoxia muscle sympa-
thetic activity and its regulation are limited to either short-term
exposures lasting no more than 30 min or long-term exposures
lasting weeks or months. During short-term hypoxia, vascular
sympathetic activity is elevated 20126%, whereas baroreex
responsiveness is unchanged or slightly augmented (12, 13,
48). During long-term exposure to hypoxia, vascular sympa-
thetic activity is elevated 200300% (6, 15). Little is known
regarding baroreex function after long-term hypoxia. Based
on these data, it is reasonable to assume that the dose-response
relation between hypoxia and MSNA follows a simple thresh-
old model. However, the current data, as well as previous data
from our laboratory, show that MSNA and sympathetic barore-
ex responsiveness decline to prehypoxia levels during the
initial stages of VAH. Thus, taking all data together, it would
appear that the hypoxia and MSNA relation displays hormesis
(23). Although it is difcult to explain this nding, it must be
considered that profound physiological adjustments are occur-
ring during this period of time. The sensitivity of arterial
chemoreceptors to PO
2
initially declines and then is augmented
along with the sensitivity of the central nervous system (31).
Concurrently, there are changes in circulating factors that
effect sympathetic activity, such as a decline in angiotensin II
(39, 41, 49).
In humans, both cardiovagal and sympathetic baroreex
responsiveness are largely unaffected by transient (25 min)
hypoxia (4, 5, 7, 12, 13). However, when the hypoxic stimulus
is prolonged to an hour, cardiovagal baroreex responsiveness
declines (35). Our data demonstrate that when the hypoxic
stimulus is prolonged to 8 h, allowing for short-term VAH,
then not only does cardiovagal baroreex tend to decline, but
the responsiveness of the sympathetic limb declines as well.
Similar interactions between autonomic reex loops have been
reported, even during acute exposure. For example, Somers
and coworkers (38) examined the effect of baroreceptor stim-
ulation on peripheral chemoreex responses during transient
hypoxia. They show that chemoreceptor-mediated responses
are blunted during baroreex activation by phenylephrine.
With the addition of ultrasound imaging during reex per-
turbation, we were able to examine two broad components of
the reex arc: 1) the mechanical, which is an estimate of the
ability of barosensory vessels to transduce arterial pressure
changes into vessel stretch; and 2) the neural, which encom-
passes afferent, central, and efferent neural function. In our
study volunteers, the carotid diameter increased during hyp-
oxia. Because the distensibility decreases with increasing ar-
terial diameter (27), it is reasonable to presume that the
Fig. 4. Integrated cardiovagal (black bars) and sympathetic (gray bars) barore-
ex responsiveness before, during, and after isocapnic hypoxia (Hx). Data are
presented as means SE. *P 0.05 vs. previous time point. AIU, arbitrary
integrated unit.
Fig. 5. The mechanical component (black bars) and neural component (gray
bars) of the sympathetic baroreex arc before hypoxia, after 1 and 7 h of
poikilocapnic hypoxia, and after return to room air. Data are presented as
means (numbers shown in bars) SE (numbers shown in parentheses). *P
0.05 vs. prehypoxia.
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transduction of arterial pressure into the carotid stretch may be
diminished during hypoxia when the carotid diameter is
greater, leading to a reduction in baroreex responsiveness.
However, we did not observe any systematic decline in the
mechanical component during the hypoxic exposure. This
could be due to several factors. First, the dilation observed is
likely due to endothelium-dependent (24, 44) or -independent
(32) ow-mediated processes. Thus, the increased distensibil-
ity associated with vascular smooth muscle relaxation may
offset the stretch-related decrease in distensibility. Second,
estimates of arterial distensibility are often derived from data
measured while arterial pressure and diameter are in a rela-
tively steady state. As we have shown before, these pulsatile
estimates do not reect the dynamic pressure-diameter inter-
action that occurs during acute baroreex perturbations (18).
Lastly, the fact that we used peripheral blood pressure esti-
mates rather than carotid pressure estimates may induce some
systematic error. The use of peripheral pressure estimates in
determining central vascular characteristics has been criticized
in the past (28). However, a direct comparison between the
central and peripheral pressure-based estimates has shown that
the use of tonometry-derived carotid pressures makes no dif-
ference in the estimate of reex relations (25). This may be
due, in part, to inherent problems regarding the valid calibra-
tion of the tonometric signals (47).
Although the reduction in sympathetic baroreex respon-
siveness was not related to a decline in the mechanical com-
ponent in our volunteers, the neural component was reduced
during hypoxia, particularly after acclimatization. This may be
due to a reduction in baroreceptor discharge rate (baroreceptor
resetting) in response to the chronic stretch of the barosensory
vessels. However, the receptor resetting is an acute phenome-
non in response to the rapid and large changes in arterial blood
pressure (20). A chronic stimulation of receptors does not lead
to further reductions in afferent neural activity (20). Therefore,
the difference in baroreex responsiveness at 1 and 7 h of
hypoxia cannot be explained by a resetting of the barorecep-
tors. Moreover, the acute reduction in baroreceptor discharge
rate is not sustained when the stimulus is removed (20). The
reduced medullary transduction of baroreceptor afferent activ-
ity into sympathetic efferent activity could also underlie the
depressed neural component of the sympathetic limb of the
baroreex. Endogenous nitric oxide production, which can
increase during hypoxia, has been shown to depress the barore-
ex afferent responsiveness in animals (22). Although a reduc-
tion in afferent responsiveness should lead to a reduction in the
neural component for both cardiovagal and sympathetic limbs,
our data show a reduction in only the sympathetic limb. This
may be due in part to the inhibitory effects of nitric oxide on
sympathetic efferent outow (36). In addition, the medullary
cardiovagal and sympathetic efferent outow may be differen-
tially regulated as demonstrated by Ma and colleagues (21).
Taken together, these previous ndings may explain the selec-
tive reduction in integrated sympathetic baroreex responsive-
ness seen in our data. However, our data cannot directly
address this possibility.
It is plausible that the hyperventilation per se may have
reduced baroreex responsiveness. Using a closed-loop ap-
proach to characterize baroreex function (frequency domain
analysis), Van De Borne and colleagues (45) reported that
isocapnic hyperventilation was associated with a decline in the
-index between systolic blood pressure and MSNA. This
decline suggests that hyperventilation alone can reduce barore-
ex modulation of vascular sympathetic activity (45). Con-
versely, using an open-loop approach (modied Oxford) Hal-
liwill et al. (13) demonstrated that hyperventilation had no
affect on baroreex responsiveness. Therefore, it remains un-
clear as to whether hyperventilation per se can reduce barore-
ex function, although it is important to note that neither Van
De Borne et al. (45) nor Halliwill et al. (13) reported that
hyperventilation affected mean MSNA levels.
Given that baroreex responsiveness and sympathetic activ-
ity were not consistently related within individuals, it is un-
likely that the decline in sympathetic activity is primarily
mediated through the baroreex. This appears to be true during
transient hypoxic exposure as well (12). However, our data
cannot address the possibility that, although blunted, the
baroreex inhibited sympathetic activity to some extent. The
effect of angiotensin II on MSNA should be considered. Acute
hypoxia has been shown to reduce arterial angiotensin II
concentrations (39, 41, 49). Since angiotensin II stimulates
sympathetic outow from the medulla, the hypoxia-mediated
decline in angiotensin II may play a role in reducing sympa-
thetic activity during VAH. However, since we did not collect
blood for analysis, our data cannot address this possibility.
It is interesting to note that upon the cessation of the hypoxic
stimulus, the forearm vascular resistance remained below pre-
exposure levels, whereas the carotid diameters returned to
baseline values. Both peripheral and cerebral blood ow ve-
locities increase during poikilocapnic hypoxia. However, fore-
arm blood ow remains elevated after hypoxia (42), whereas
cerebral blood ow returns to preexposure levels after expo-
sure (30). With this in mind, the reduced forearm vascular
resistance is likely due to reduced vascular sympathetic activity
along with enhanced nitric oxide production causing vasodila-
tion in the more muscular peripheral radial and ulnar arteries
and arterioles. In contrast, the blood ow through the carotid
artery likely returned to baseline levels, reducing shear stress.
Moreover, the less muscular artery is likely less affected by the
reduced vascular sympathetic outow.
Limitations
A consistent subject of debate in the study of autonomic
regulation is the proper metric for expressing cardiovagal
baroreex responsiveness. Because of the inverse, curvelinear
relation between R-R interval and heart rate, a rather large
change in R-R interval can result in a small change in heart
rate, potentially obscuring changes in autonomic regulation.
Since heart rate is linearly related to cardiac output, and thus
blood pressure, this metric may be most appropriate when
examining the ability of the baroreex to buffer changes in
arterial blood pressure. However, in the current investigation,
our experimental question was related to changes in the auto-
nomic control of afferent neural activity. Therefore, since
carotid afferent frequency is linearly related to R-R interval in
cats and humans (3, 19), this metric seems most appropriate for
this investigation.
Given that the microelectrode was removed after 1 h of
hypoxia and then replaced before the 7-h hypoxia measure-
ment, the reproducibility of MSNA is an important issue. First,
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it should be noted that the key dependent variable, sympathetic
baroreex responsiveness, is based on the change in MSNA
rather than the absolute levels of MSNA. Nonetheless, intra-
individual MSNA is highly reproducible, even when measured
several years apart (9). Moreover, MSNA recorded in the leg is
strongly associated with MSNA measured simultaneously in
the arm (33, 40, 46). Taken together, this should provide
assurance that our measurements of both steady-state MSNA
and sympathetic baroreex responsiveness, taken in the same
proximity but different site within the peroneal nerve, can be
compared quantitatively.
Conclusions
These data provide evidence that baroreex function is
inhibited during the initial stages of VAH. Furthermore, by the
addition of continuous ultrasound images of the carotid artery
during baroreceptor stimulation, we have been able to provide
data that strongly suggest that the reduction in baroreex
responsiveness is likely due to changes in the medullary
afferent-efferent integration, possibly associated with ventila-
tory acclimatization to hypoxia. Although our data do not
directly address this hypothesis, taken together with previous
data in animals, it is likely. Furthermore, in contrast to previous
work that has only examined the chemoreceptor-baroreceptor
interactions before acclimatization, our work suggests that this
transition period during hypoxia encompasses a unique inte-
gration between chemoreceptor, baroreceptor, and end-organ
function.
GRANTS
This research was supported by National Heart, Lung, and Blood Institute
Grant HL-072648-01A1.
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