J Appl Physiol 102: 2071–2076, 2007;
doi:10.1152/japplphysiol.01258.2006.
Point:Counterpoint: Release of an endothelium-derived vasoconstrictor and
RhoA/Rho kinase-mediated calcium sensitization of smooth muscle cell
contraction are/are not the main effectors for full and sustained hypoxic
pulmonary vasoconstriction
POINT: RELEASE OF AN ENDOTHELIUM-DERIVED
VASOCONSTRICTOR AND RHOA/RHO KINASE-MEDIATED
CALCIUM SENSITIZATION OF SMOOTH MUSCLE CELL
CONTRACTION ARE THE MAIN EFFECTORS FOR FULL AND
SUSTAINED HYPOXIC PULMONARY VASOCONSTRICTION
The ability of an organism, or an organ therein, to respond
appropriately to episodes of hypoxia is a key evolutionary
determinant. In mammalian lungs, the response to acute hyp-
oxia involves constriction of the intrapulmonary arteries (IPA)
in the hypoxic region of the lung. This response, termed
hypoxic pulmonary vasoconstriction (HPV), effectively
matches regional perfusion of the lung to regional ventilation
and constitutes a pivotal homeostatic mechanism (25). HPV
has, therefore, been the subject of intensive research for many
years, although little consensus exists regarding its underlying
sensor(s) and effector(s) mechanisms. Indeed, heated debates
abound once one mentions trigger words and phrases such as
“redox,” “reactive oxygen species,” “potassium channels,”
etc., in relation to HPV (1, 28). This article focuses attention on
another incendiary trigger phrase, namely “the endothelium.”
To consider the role of the endothelium in HPV, we must
first define the hypoxic constrictor response itself. In vivo and
in isolated blood-perfused lungs, HPV is usually immediate in
onset and sustained. In contrast, HPV in isolated arteries is
often biphasic, consisting of an initial transient constrictor
response superimposed on a more slowly developing, sustained
contraction (10). It has been argued that the latter, sustained,
response is more physiologically relevant as it would appear
important that HPV be a sustained response so that the redi-
rection of blood flow in response to regional hypoxia is
maintained for the duration of the hypoxic episode. It is
important to note that although sustained HPV in isolated
arteries is often referred to as “phase 2,” this does not mean
that it is secondary to the transient phase of HPV. When
pharmacological agents are used to block selectively the tran-
sient (i.e., “phase 1”) response (5, 20), it becomes apparent that
sustained HPV in isolated arteries is immediate in onset, akin
to that observed in perfused lungs and in vivo. In the review of
Aaronson et al. (2), the authors tabulated the results of studies
where the endothelium dependency of sustained HPV was
examined. The final score of studies finding an endothelium
dependency, be it partial or whole, versus those that found no
role for the endothelium was 15 to 4. Given this convincing
scoreline and the fact that it was spread over six species, I am
sure that a betting person would place their wager on the
proendothelium side of the debate!
Since the endothelial dependency of sustained HPV does not
appear to be attributable to a reduction in the release of
relaxing factors (2, 22), the case for an endothelium-derived
constrictor factor(s) (EDCF) is compelling. Indeed, several
studies have reported that an EDCF is released during HPV,
although its precise identity remains elusive (5, 7, 21, 24). One
http://www. jap.org 8750-7587/07 $8.00 Copyright © 2007 the American Physiological Society
Point:Counterpoint
possible candidate for this EDCF would be endothelin-1 (ET-
1), and it has been postulated that ET-1 may play a priming role
in some species (12, 22). However, ET-1 receptor antagonists are
more often than not found to be ineffective in blocking HPV (2,
22) and ET-1 would appear to be a more likely an “agent
provocateur” in chronic pulmonary hypertension (4).
Although the identity of the EDCF responsible for sustained
HPV is unclear, more clarity exists concerning the mechanism
by which it elicits contraction of the pulmonary vascular
smooth muscle. It is apparent that the two principal determi-
nants of smooth muscle tone, namely cytosolic Ca2⫹ ([Ca2⫹]i)
and the sensitivity of the contractile apparatus to Ca2⫹, are
Downloaded from jap.physiology.org on May 6, 2007
elevated during sustained HPV (18). Specifically, following the
transient rises in tension and [Ca2⫹]i associated with phase 1 of
HPV, [Ca2⫹]i remains at a constant level (above that prior to
hypoxia) while tension continues to rise (18, 20). The lack of
correlation between tension and [Ca2⫹]i during sustained HPV
is a classic example of Ca2⫹ sensitization. Endothelial denu-
dation was found to abolish sustained HPV independent of any
effect on the level of smooth muscle [Ca2⫹]i (17). These
observations are wholly consistent with the elevation in smooth
muscle [Ca2⫹]i being an inherent property of pulmonary vas-
cular smooth muscle, whereas the Ca2⫹ sensitization observed
during sustained HPV is the purview of the EDCF.
At the time of this initial observation, one potential mediator
of the Ca2⫹ sensitization observed during HPV, protein kinase
C, was eliminated and it was not until the availability of the
specific inhibitor of Rho-associated coiled coil-forming serine/
threonine kinase (Rho kinase), Y-27632, that the activation of
Rho kinase was first promulgated as a pivotal step in sustained
HPV (19). Since the first isolation of Rho kinase (11), it has
become apparent that, upon activation by the binding of the
small monomeric G-protein RhoA, Rho kinase inhibits myosin
light chain phosphatase (MLCP), resulting in Ca2⫹-indepen-
dent contraction of vascular smooth muscle. Since this initial
report, converging lines of evidence have pinpointed Rho
kinase activation and subsequent inhibition of MLCP (27) as
being a vital step in the initiation and maintenance of increased
pulmonary artery pressure in response to acute hypoxia (6, 23),
chronic hypoxia (6, 8, 13, 15), and pulmonary hypertensive
states per se (3, 13, 14). Moreover, Rho-kinase inhibitors
appear to show much promise as potential therapeutic agents
in pulmonary hypertension (9). It is, therefore, possible that
the activation of Rho kinase during sustained HPV repre-
sents the bridge between physiological HPV (i.e., that which
occurs in response to brief and reversible hypoxia) and the
pathophysiological consequences of chronic hypoxia.
It would seem logical to assume that since sustained HPV
and the associated Ca2⫹ sensitization are both endothelium
dependent then the activation of Rho kinase during hypoxia
should be similarly endothelium dependent. However, it was
reported that hypoxia can directly activate Rho kinase in
2071
Point:Counterpoint
2072
cultured pulmonary artery smooth muscle cells (PASMC; Ref.
26). At first glance, this appears to scupper any idea of an
EDCF-mediated increase in Rho kinase activation during sus-
tained HPV. However, the increase in Rho kinase activity in
cultured PASMC was a rather meager ⬃20% after 40 min of
hypoxia, rising to a rather disappointing ⬃40% after 1 h. This
is in stark contrast to the rather impressive ⬃260% increase in
Rho kinase activity recently reported in isolated pulmonary
arteries of the rat during sustained HPV (16). Moreover, this
activation of Rho kinase in rat pulmonary arteries was endo-
thelium dependent and in endothelium-denuded arteries the
increase in Rho kinase activity was similar (⬃40%) to that
reported for cultured PASMC. Furthermore, the translocation
of RhoA to the plasma membrane of the pulmonary smooth
muscle, a causal event in the activation of Rho kinase, was
similarly dependent on the presence on an intact endothelium.
In closing, the case for the EDCF-RhoA/Rho kinase-Ca2⫹-
sensitization axis being responsible for the maintenance of sus-
tained HPV is solid. My most worthy opponent will probably try
to convince the reader that the endothelium merely “modulates”
the hypoxic constrictor response. However, when disruption of
said axis results in removal of the hypoxic constrictor response,
then “modulate” must surely be changed to “mediate”!
REFERENCES
1. Aaronson PI, Robertson TP, Knock GA, Becker S, Lewis TH, Snetkov
V, Ward JPT. Hypoxic pulmonary vasoconstriction: mechanisms and
controversies. J Physiol 570: 53–58, 2006.
2. Aaronson PI, Robertson TP, Ward JPT. Endothelium-derived media-
tors and hypoxic pulmonary vasoconstriction. Resp Physiol Neurobiol
132: 107–120, 2002.
3. Abe K, Shimokawa H, Morikawa K, Uwatoku T, Oi K, Matsumoto Y,
Hattori T, Nakashima Y, Kaibuchi K, Sueishi K, Takeshit A. Long-
term treatment with a Rho-kinase inhibitor improves monocrotaline-
induced fatal pulmonary hypertension in rats. Circ Res 94: 385–393, 2004.
4. Channick RN, Sitbon O, Barst RJ, Manes A, Rubin LJ. Endothelin
receptor antagonists in pulmonary arterial hypertension. J Am Coll Cardiol
43: 62S– 67S, 2004.
5. Evans AM, Dipp M. Hypoxic pulmonary vasoconstriction: cyclic aden-
osine diphosphate-ribose, smooth muscle Ca2⫹ stores and the endothe-
lium. Respir Physiol Neurobiol 132: 3–15, 2002.
6. Fagan KA, Oka M, Bauer NR, Gebb SA, Ivy DD, Morris KG,
McMurtry IF. Attenuation of acute hypoxic pulmonary vasoconstriction
and hypoxic pulmonary hypertension in mice by inhibition of Rho-kinase.
Am J Physiol Lung Cell Mol Physiol 287: L656 –L664, 2004.
7. Gaine SP, Hales MA, Flavahan NA. Hypoxic pulmonary endothelial
cells release a diffusible contractile factor distinct from endothelin. Am J
Physiol Lung Cell Mol Physiol 274: L657–L664, 1998.
8. Hyvelin JM, Howell K, Nichol A, Costello CM, Preston RJ, McLough-
lin P. Inhibition of Rho-kinase attenuates hypoxia-induced angiogenesis in
the pulmonary circulation. Circ Res 97: 185–191, 2005.
9. Ishikura K, Yamada N, Ito M, Ota S, Nakamura M, Isaka N, Nakano
T. Beneficial acute effects of Rho-kinase inhibitor in patients with pul-
monary arterial hypertension. Circ J 70: 174 –178, 2006.
10. Leach RM, Robertson TP, Twort CH, Ward JP. Hypoxic vasoconstric-
tion in rat pulmonary and mesenteric arteries. Am J Physiol Lung Cell Mol
Physiol 266: L223–L231, 1994.
11. Leung T, Manser E, Tan L, Lim L. A novel serine/threonine kinase
binding the Ras-related RhoA GTPase which translocates the kinase to
peripheral membranes. J Biol Chem 270: 29051–29054, 1995.
12. Liu Q, Sham JS, Shimoda LA, Sylvester JT. Hypoxic constriction of
porcine distal pulmonary arteries: endothelium and endothelin depen-
dence. Am J Physiol Lung Cell Mol Physiol 280: L856 –L865, 2001.
13. Nagaoka T, Fagan KA, Gebb SA, Morris KG, Suzuki T, Shimokawa
H, McMurtry IF, Oka M. Inhaled Rho kinase inhibitors are potent and
selective vasodilators in rat pulmonary hypertension. Am J Respir Crit
Care Med 171: 494 – 499, 2005.
J Appl Physiol • VOL
14. Nagaoka T, Gebb SA, Karoor V, Homma N, Morris KG, McMurtry
IF, Oka M. Involvement of RhoA/Rho kinase signaling in pulmonary
hypertension of the fawn-hooded rat. J Appl Physiol 100: 996 –1002, 2006.
15. Nagaoka T, Morio Y, Casanova N, Bauer N, Gebb S, McMurtry I,
Oka M. Rho/Rho kinase signaling mediates increased basal pulmonary
vascular tone in chronically hypoxic rats. Am J Physiol Lung Cell Mol
Physiol 287: L665–L672, 2004.
16. Robertson T, Hashmi-Hill M, Vandenplas ML, Lewis SJ. Endothe-
lium-dependent activation of Rho-kinase during hypoxic pulmonary va-
soconstriction in rat intrapulmonary arteries (Abstract). FASEB J 19:
A1277, 2005.
17. Robertson TP, Aaronson PI, Ward JPT. Ca2⫹ sensitization during
sustained hypoxic pulmonary vasoconstriction is endothelium dependent.
Am J Physiol Lung Cell Mol Phys 284: L1121–L1126, 2003.
18. Robertson TP, Aaronson PI, Ward JPT. Hypoxic vasoconstriction and
intracellular Ca2⫹ in pulmonary arteries: evidence for PKC-independent
Ca2⫹-sensitization. Am J Physiol Heart Circ Physiol 37: H301–H307, 1995.
19. Robertson TP, Dipp M, Ward JPT, Aaronson PI, Evans AM. Inhibi-
tion of sustained hypoxic vasoconstriction by Y-27632 in isolated intrapul-
monary arteries and perfused lung of the rat. Br J Pharmacol 131: 5–9,
2000.
20. Robertson TP, Hague D, Aaronson PI, Ward JPT. Voltage-indepen-
dent calcium entry in hypoxic pulmonary vasoconstriction of intrapulmo-
nary arteries of the rat. J Physiol 525: 669 – 680, 2000.
Downloaded from jap.physiology.org on May 6, 2007
21. Robertson TP, Ward JPT, Aaronson PI. Hypoxia induces the release of
a pulmonary-selective, Ca2⫹-sensitising, vasoconstrictor from the per-
fused rat lung. Cardiovasc Res 50: 145–150, 2001.
22. Shimoda LA, Sham JS, Liu Q, Sylvester JT. Acute and chronic hypoxic
pulmonary vasoconstriction: a central role for endothelin-1? Respir
Physiol Neurobiol 132: 93–106, 2002.
23. Sylvester JT. The tone of pulmonary smooth muscle: ROK and Rho
music? Am J Physiol Lung Cell Mol Physiol 287: L624 –L630, 2004.
24. Talbot NP, Robbins PA, Dorrington KL. Release by hypoxia of a
soluble vasoconstrictor from rabbit small pulmonary arteries. Br J Anaesth
91: 592–594, 2003.
25. Von Euler US, Liljestrand G. Observations on the pulmonary arterial
blood pressure in the cat. Acta Physiol Scand 12: 301–320, 1946.
26. Wang Z, Jin N, Ganguli S, Swartz DR, Li L, Rhoades RA. Rho-kinase
activation is involved in hypoxia-induced pulmonary vasoconstriction.
Am J Respir Cell Mol Biol 25: 628 – 635, 2001.
27. Wang Z, Lanner MC, Jin N, Swartz D, Li L, Rhoades RA. Hypoxia
inhibits myosin phosphatase in pulmonary arterial smooth muscle cells:
role of Rho kinase. Am J Respir Cell Mol Biol 29: 465– 471, 2003.
28. Ward JP, Weir EK, Archer SL. Hypoxic pulmonary vasoconstriction
is/is not mediated by increased production of reactive oxygen species.
J Appl Physiol 101: 993–999, 2006.
Tom P. Robertson
Department of Physiology and Pharmacology
Institute of Comparative Medicine
College of Veterinary Medicine
The University of Georgia
Athens, Georgia
e-mail: troberts@vet.uga.edu
COUNTERPOINT: RELEASE OF AN ENDOTHELIUM-DERIVED
VASOCONSTRICTOR AND RHOA/RHO KINASE-MEDIATED
CALCIUM SENSITIZATION OF SMOOTH MUSCLE CELL
CONTRACTION ARE NOT THE MAIN EFFECTORS FOR FULL
AND SUSTAINED HPV
The great tragedy of Science—the slaying of a beautiful hypo-
thesis by an ugly fact
Thomas H. Huxley (1825–1895)
With a simple experiment 15 years ago, Jane Madden et al.
(9) slayed the beautiful (then unborn) hypothesis that a (still
mysterious) vasoconstrictor secreted from the endothelium in
response to hypoxia causes hypoxic pulmonary vasoconstric-
102 • MAY 2007 • www.jap.org

Fig. 1. Resistance pulmonary artery (PA) isolated smooth muscle cells (SMC), but not systemic arterial SMC, are able to fully contract to hypoxia, suggesting
that the endothelium is not essential (from Ref. 9 with permission).
tion (HPV) via the Rho-kinase pathway (1, 21). Freshly
isolated (and nondedifferentiated) pulmonary artery smooth
muscle cells (PASMC), but not systemic arterial SMC, were
able to contract to hypoxia in vitro (9); no endothelium, no
mysterious factors needed (Fig. 1). There is nothing ugly to
this experiment, although it might appear ugly in the eyes of
our opponent. We could rest our case right here, but let’s
continue.
Our job becomes even easier because of the way the debat-
able statement is worded: “. . . .the main effectors. . . .” being
the key words. Many mechanisms contribute to full HPV, from
the nervous to the endocrine system and multiple local medi-
ators. But the main effector, the cornerstone of an HPV theory,
has to support all the fundamental features of HPV. To win the
argument, we simply need to prove that our opponent’s theory
is in conflict with only some of these.
HPV is intrinsic to the pulmonary circulation. While the
resistance PAs constrict to hypoxia (to maintain ventilation-
perfusion matching), the systemic arteries do not, and often
dilate, increasing blood delivery to the hypoxic/ischemic tis-
sues. This means that a mechanism for HPV has to explain its
selectivity to the resistance PAs. Similarly, it needs to be
specific to hypoxia and ideally not shared by the many path-
ways involved in nonspecific PA and systemic artery constric-
tions. For example, HPV cannot be explained by a mechanism
involved in the vasoconstriction to prostanoids or phenyleph-
rine, which constrict proximal, resistance PAs and systemic
arteries in a similar manner. Based on these, here are three
points that support our case.
Point 1: An argument that our opponent will use is that the
well-documented increase in [Ca2⫹]i caused by hypoxia in
PASMC is small and transient, not enough to cause full and
sustained contraction (1). Therefore, he will claim, a “boost”
from an endothelium-derived vasoconstrictor and an increase
in Ca2⫹ sensitivity, mediated by Rho kinase, is necessary. This
argument is often used to justify pretone strategies, i.e., expo-
sure to a nonspecific vasoconstrictor prior to and during hyp-
oxia. The breakdown of HPV to phase I and II [phase II is the
endothelium- and Rho kinase-dependent phase (1)] is not a
J Appl Physiol • VOL
Point:Counterpoint
2073
Downloaded from jap.physiology.org on May 6, 2007
universal feature of HPV. For example, it is seen in the isolated
PA model, but not in the perfused lung or in vivo models. It
also appears that pretone is required in some but not all
laboratories. For example, in our hands, rat resistance PAs can
have full HPV without pretreatment (3). Although species
differences might exist, it is ironic that in the cat Pas [the
model in which HPV was first described (24)], no preconstric-
tion is required for HPV (7, 10). Furthermore, in the cat, rat,
lamb, dog, and other species (3, 6, 8, 10, 25), full HPV can be
observed in isolated PAs effectively denuded from endothe-
lium (Fig. 1B). While methodological differences can be de-
bated in the isolated PA model, our strongest argument comes
from the freshly isolated PASMC model (9). These cells can
effectively contract within a few minutes of exposure to phys-
iological hypoxia and without the need of pretreatment with
vasoconstrictors (Fig. 1A); obviously the increase in [Ca2⫹]i in
this case is by itself enough for effective contraction.
Our opponent’s theory has a conflict with this observation.
This is not the case with competing theories. Take for example
the theory describing an oxygen sensor within the PASMC (the
mitochondria, or the balance of redox couples), which in
response to changes in PO2, alters the production of a mediator
(for example ROS, whether increased or decreased), which
regulates the function of an effector (for example, voltage-
gated K⫹ channels, Kv) (15, 26). No need for endothelium,
mysterious factors, pretone requirements, etc. In its simplicity,
it is also shared by many other oxygen-sensing systems in the
body, including neurons or neuroendocrine cells (26). The Kv
channel inhibition in isolated PASMC occurs within minutes
from exposure to physiological hypoxia (19). The resulting
depolarization leads to increased [Ca2⫹]i and contraction. If
one keeps these cells in hypoxia for days (chronic hypoxia), he
will observe a sustained Kv channel inhibition and a sustained
increase in [Ca2⫹]i (18). Interestingly this does not occur in
systemic arterial SMC treated identically (17, 27). This sus-
tained mechanism, supported by molecular events that take
place in chronic hypoxia, has been proposed as the basis of
hypoxic pulmonary hypertension (11, 15, 26).
A proposed mechanism for HPV has to explain PA constric-
tion but also lack of effects or (ideally) dilatation of systemic
102 • MAY 2007 • www.jap.org
Point:Counterpoint
2074
arteries. Furthermore, it should not be a part of any nonspecific
vasoconstrictor response. The vasoconstrictor-Rho kinase the-
ory suffers badly here.
Point 2: The inhibition of HPV by Rho-kinase inhibitors (in
a manner similar to the inhibition caused by lack of endothe-
lium) is used by our opponent to defend his point (1). But
Rho-kinase inhibitors inhibit angiotensin II and KCl-induced
constriction in a manner similar to HPV (5). In contrast,
blocking Kv channels pharmacologically (3) or molecularly (2,
20) inhibits HPV in isolated PASMC, arteries, and lungs, but
not the response to other constrictors like phenylephrine.
Point 3: And here is the worst part: Rho-kinase inhibitors
dilate many systemic arteries, even cerebral arteries (22). In
fact, they have been proposed as potential treatment for systemic
hypertension (23). The systemic administration of Rho-kinase
inhibitors is limited in pulmonary hypertension (PHT) because of
significant systemic hypotension (16). In an attempt to bypass the
systemic effects, inhaled Rho-kinase inhibitors (fasudil) are used
to treat experimental PHT (16).
Now look at just one example of a drug (rotenone) that
blocks the mitochondrial complex I, thus proximally inhibiting
the mitochondria-Kv channel oxygen-sensing system that we
discussed above. In a system where a rat lung and kidney are
perfused in series, rotenone mimics hypoxia and constricts the
pulmonary while it dilates the renal circulation; this is con-
firmed in endothelium-denuded vascular rings and in isolated
SMC where rotenone inhibits Kv current in PA but activates
Kv current in renal artery SMC (13). This is explained by the
fact that mitochondria appear to be different in the PA vs.
systemic arterial SMC (13). Furthermore, dichloroacetate
(DCA), a drug that inhibits a mitochondrial enzyme (pyruvate
dehydrogenase kinase), when given systemically in several
models of PHT, significantly decreases pulmonary vascular
resistance, without affecting systemic arterial pressure (4, 12,
14). While Rho-kinase inhibitors appear to be nonspecific for
the pulmonary circulation, targeting the mitochondrial-Kv
channel axis selectively inhibits HPV and reverses PHT, spar-
ing systemic vessels.
HPV remains mysterious and elegant. In a future debate, we
could defend the mitochondria-ROS-Kv channel theory. For
this one, we only had to reject our opponent’s theory; a much
easier job. Now let’s clarify something. Our opponent’s work
is brilliant. We have been his and his team’s students over the
years. Their most recent review (1) is one of the best, most
balanced, and comprehensive reviews we have seen in this
field. Their original description of this pathway remains one of
the most influential in the field (21). The Rho-kinase pathway
is an important one for pulmonary vascular biology; but it is
not THE one for HPV.
If you are out to determine what it is that makes a Ferrari
accelerate so differently than a Fiat, you have to first accept
that there is probably more than one reason. However, if you
deflate the tires and the Ferrari does not run, you can’t con-
clude that you discovered what makes the Ferrari a Ferrari;
tires are equally important for all cars, like Rho kinase is
important in all blood vessels.
To reveal the elegance and uniqueness of a Ferrari, you have
to dig further, deep inside its engine, deep into the mitochon-
dria.
J Appl Physiol • VOL
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Calcium sensitization of smooth muscle mediated by a Rho-associated
protein kinase in hypertension. Nature 389: 990 –994, 1997.
24. Von euler US, Liljestrand G. Observations on the pulmonary arterial
blood pressure in the cat. Acta Physiol Scand 12: 301–320, 1946.
25. Wang Y, Coe Y, Toyoda O, Coceani F. Involvement of endothelin-1 in
hypoxic pulmonary vasoconstriction in the lamb. J Physiol 482: 421–434, 1995.
26. Weir EK, Lopez-Barneo J, Buckler KJ, Archer SL. Acute oxygen-
sensing mechanisms. N Engl J Med 353: 2042–2055, 2005.
27. Yuan XJ, Goldman WF, Tod ML, Rubin LJ, Blaustein MP. Hypoxia
reduces potassium currents in cultured rat pulmonary but not mesenteric
arterial myocytes. Am J Physiol Lung Cell Physiol 264: L116 –L123, 1993.
Gaël Y. Rochefort
Evangelos D. Michelakis
Pulmonary Hypertension Program
University of Alberta
Edmonton, Canada
e-mail: emichela@cha.ab.ca
REBUTTAL FROM DR. ROBERTSON
Two words came to mind as I read my opponent’s side of the
argument: “smoke” and “mirrors.” To quote my esteemed
opponents: “our strongest argument comes from the freshly
isolated PASMC model.” While I do not regard these experi-
ments as “ugly,” they may have more than a few unsightly
blemishes! The observation that enzymatically dispersed
PASMCs constrict to hypoxia in no way negates a primary role
for an endothelium-dependent Ca2⫹ sensitization in the intact
artery or in the whole animal. To infer that the PASMC is the
sole repository for the mechanism(s) of HPV as it can constrict
to hypoxia, with complete disregard to the myriad of cellular
changes that likely occur during its rather traumatic isolation,
is one inference too far!
I must also take issue with my opponent’s argument that
HPV “cannot be explained by a mechanism involved in the
vasoconstriction to prostanoids or phenylephrine.” I must
admit to finding this argument rather baffling. Could not the
specificity, with respect to HPV, be attributed to an endothe-
lium-derived constrictor factor released locally within the lung,
and/or one that acts solely within the pulmonary circulation
rather than in the signal transduction pathway it activates?
It is interesting that my opponents state that “methodologi-
cal differences can be debated in the isolated PA model.”
Indeed they can and since my opponents state that “in our
hands, rat resistance PAs can have full HPV” (1), this seems
like a suitable point to debate some methodological differ-
ences! In their work, the PAs were ⬃74 ␮m in diameter and
had a resting tension of 700 mg (⬃6.9 mN). With a generous
allowance of between 1 and 2 mm vessel length, for an artery
of this diameter, the equivalent transmural pressure would
probably lie between ⬃500 and ⬃1,000 mmHg compared with
15–30 mmHg reported from other laboratories (2–9). More-
over, the resultant constrictor responses in this report were,
J Appl Physiol • VOL 102 • MARCH 2007 •
Point:Counterpoint
2075
perhaps unsurprisingly, given the “resting” conditions, rather
small (⬃100 mg or ⬃1 mN) compared with those reported by
other investigators using small PAs (⬃30 mN; Refs. 2, 4, 6, 7).
In the laboratories in which I have had the privilege to work, a
pulmonary artery with a resting tension of 7 mN and a
subsequent constrictor response of 1 mN would have been a
candidate for life support measures!
Finally, HPV (like a Ferrari) is multifactorial. It requires a
spark (the mitochondria and increases in cytosolic Ca2⫹), fuel
(glucose for glycolysis), and an accelerator (Rho kinase). But
one thing is missing in this Ferrari. . .oh yes, the driver—
something to put the pedal to the metal when it’s really
needed. . . .now I wonder what that could be?
REFERENCES
1. Archer SL, Wu XC, Thebaud B, Nsair A, Bonnet S, Tyrrell B, Mc-
Murtry MS, Hashimoto K, Harry G, Michelakis ED. Preferential ex-
pression and function of voltage-gated, O2-sensitive K⫹ channels in
resistance pulmonary arteries explains regional heterogeneity in hypoxic
pulmonary vasoconstriction: ionic diversity in smooth muscle cells. Circ
Res 95: 308 –318, 2004.
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2. Dipp M, Evans AM. Cyclic ADP-ribose is the primary trigger for hypoxic
pulmonary vasoconstriction in the rat lung in situ. Circ Res 89: 77– 83,
2001.
3. Jones RD, Morice AH. The effect of the nitric oxide synthase inhibitor
Ngamma-nitro-L-argine methyl ester on hypoxic pulmonary vasoconstric-
tion. Eur J Pharmacol 402: 111–117, 2000.
4. Leach RM, Hill HM, Snetkov VA, Robertson TP, Ward JPT. Divergent
roles of glycolysis and the mitochondrial electron transport chain in hypoxic
pulmonary vasoconstriction of the rat: identity of the hypoxic sensor.
J Physiol 536: 211–224, 2001.
5. Morecroft I, Loughlin L, Nilsen M, Colston J, Dempsie Y, Sheward J,
Harmar A, MacLean MR. Functional interactions between 5-hydroxy-
tryptamine receptors and the serotonin transporter in pulmonary arteries.
J Pharmacol Exp Ther 313: 539 –548, 2005.
6. Robertson TP, Aaronson PI, Ward JP. Hypoxic vasoconstriction and
intracellular Ca2⫹ in pulmonary arteries: evidence for PKC-independent
Ca2⫹ sensitization. Am J Physiol Heart Circ Physiol 268: H301–H307,
1995.
7. Robertson TP, Dipp M, Ward JP, Aaronson PI, Evans AM. Inhibition
of sustained hypoxic vasoconstriction by Y-27632 in isolated intrapulmo-
nary arteries and perfused lung of the rat. Brit J Pharmacol 131: 5–9, 2000.
8. Teng GQ, Barer GR. In vitro responses of lung arteries to acute hypoxia
after NO synthase blockade or chronic hypoxia. J Appl Physiol 79: 763–
770, 1995.
9. Thomas BJ, Wanstall JC. Alterations in pulmonary vascular function in
rats exposed to intermittent hypoxia. Eur J Pharmacol 477: 153–161, 2003.
REBUTTAL FROM DRS. ROCHEFORT AND MICHELAKIS
Opening his argument, our opponent calls you, the readers,
to a gambling exercise; admittedly a safe bet. Because in one
of his reviews on the subject, only four papers found the
endothelium to be nonessential for HPV, compared to the 15
that found it to be essential (1), he asks you to bet your money
on his side of the argument. However, even in Las Vegas you
get in trouble if you play safe bets, but with weighted dice! To
support their side of the story in that review, the authors listed
(in Table 1 of that review) apparently all the papers that were
studying sustained HPV (more than 20 min of hypoxia) (1).
We do not understand the basis on which 20 min was used as
the cutoff between acute and “sustained HPV.” But, even using
this rule, several papers were omitted from that table and
argument, which, interestingly, strongly suggest a nonessential
role of the endothelium. For example, Wang et al. (3) studied
www.jap.org
Point:Counterpoint
2076
isolated lamb pulmonary arteries: even up to 120 min of
hypoxia, caused the same constriction in the vessels with and
without endothelium. In fact, the endothelium-denuded vessels
had a faster constriction to hypoxia. Jane Madden’s work
[including the paper that we discussed in our argument and in
our Fig. 1A (2)] is also not included in the 4/15 argument. We
could improve the odds more by adding papers from our group
(published before and after that review), but lucky you, you are
scientists and not gamblers.
Nevertheless, this Point:Counterpoint series proves to be
much easier to settle than expected. For example, our opponent
alerts you to the fact that we would try to downgrade the
importance of endothelial-derived factors by presenting them
as modulators of HPV. But we do not really have to. He has
done this for us, in the “gambler’s guide to HPV,” that he
offered you (2). The conclusion of this review reads: “media-
tors derived from the pulmonary vascular endothelium exert a
J Appl Physiol • VOL
powerful modulating influence on the response of the pulmo-
nary circulation to hypoxia.” We could not agree more.
In his current argument, our opponent concludes: “It is,
therefore, possible that the activation of Rho kinase during
sustained HPV represents the bridge between physiological
HPV. . . and the pathophysiological consequences of chronic
hypoxia.”
REFERENCES
1. Aaronson PI, Robertson TP, Ward JP. Endothelium-derived mediators
and hypoxic pulmonary vasoconstriction. Respir Physiol Neurobiol 132:
107–120, 2002.
2. Madden JA, Vadula MS, Kurup VP. Effects of hypoxia and other
vasoactive agents on pulmonary and cerebral artery smooth muscle cells.
Am J Physiol Lung Cell Mol Physiol 263: L384 –L393, 1992.
3. Wang Y, Coe Y, Toyoda O, Coceani F. Involvement of endothelin-1 in
hypoxic pulmonary vasoconstriction in the lamb. J Physiol 482: 421– 434,
1995.
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102 • MARCH 2007 • www.jap.org
J Appl Physiol 102: 2077–2079, 2007;
doi:10.1152/japplphysiol.01435.2006.
The following letters are in response to Point:Counterpoint:
“Release of an endothelium-derived vasoconstrictor and
RhoA/Rho kinase-mediated calcium sensitization of smooth
muscle cell contraction are/are not the main effectors for full
and sustained hypoxic pulmonary vasoconstriction” that
appears in this issue.
To the Editor: Obviously, I’m prejudiced, but evidence for the
PASMC as the main effector for HPV seems compelling, not
only because of our work (1) kindly cited by Rochefort and
Michelakis (5), but also because of work not mentioned.
Murray et al. (2) showed that fetal calf PASMCs grown on a
flexible membrane could generate tension when exposed to
hypoxia. Olson et al.’s (3), provocative study in the dorsal
aorta of the most primitive vertebrate, the cyclostome, suggests
that hypoxic vasoconstriction is phylogenetically ancient and
inherent to vascular smooth muscle. This vessel exhibited
repeatable and sustained hypoxic contractions without precon-
ditioning and without an endothelium or contributions from
other agonist pathways. Olson et al. (3) postulate that evolution
has resulted in the almost exclusive association of hypoxic
vasoconstriction to respiratory organs in the higher vertebrates
and that “it has been embellished with secondary regulatory
mechanisms” such as those from the endothelium. Tweaking,
embellishing, and reinforcing seem to be common themes in
nature and maybe Robertson’s argument for an endothelium
derived contracting factor is based on the glitz and not the grit
of HPV. Finally, Robertson (4) uses the trauma of isolation to
argue against the PASMC as the effector of HPV. If the cells
were traumatized, how could they contract to hypoxia? Isola-
tion and the resultant loss of an endothelial contractile factor
did not confer this ability upon them. In isolated arteries,
however, HPV is often lost after any manipulation that disrupts
the vessel’s functional integrity (think mechanical and/or
chemical endothelium removal).
REFERENCES
1. Madden JA, Vadula MS, Kurup VP. Effects of hypoxia and other
vasoactive agents on pulmonary and cerebral artery smooth muscle cells.
Am J Physiol Lung Cell Mol Physiol 263: L384 –L393, 1992.
2. Murray T, Chen L, Marshall B, Macarak E. Hypoxic contraction of
cultured pulmonary vascular smooth muscle cells. Am J Respir Cell Mol
Biol 3: 457– 465, 1990.
3. Olson KR, Russell MJ, Forster ME. Hypoxic vasoconstriction of cyclo-
stome systemic vessels: the antecedent of hypoxic pulmonary vasoconstric-
tion? Am J Physiol Regul Integr Comp Physiol 280: R198 –R206, 2001.
4. Robertson TP. Point: Release of an endothelium-derived vasoconstrictor
and RhoA/Rho kinase-mediated calcium sensitization of smooth muscle
cell contraction are the main effectors for full and sustained hypoxic
pulmonary vasoconstriction. J Appl Physiol. In press.
5. Rochefort GY, Michelakis ED. Counterpoint: Release of an endothelium-
derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitiza-
tion of smooth muscle cell contraction are not the main effectors for full and
sustained hypoxic pulmonary vasoconstriction. J Appl Physiol. In press.
Jane A. Madden
Medical College of Wisconsin and
Zablocki Veterans Affairs Medical Center
To the Editor: Cultured or dispersed pulmonary vascular
smooth muscle cells can constrict when subjected to hypoxia
and a very large body of evidence implicates oxygen-sensitive
K⫹ (and perhaps other) ion channels. However the question
that is ignored by the proponents of smooth muscle ion chan-
http://www. jap.org 8750-7587/07 $8.00 Copyright © 2007 the American Physiological Society
Point:Counterpoint Comments
nels is whether cells in culture are behaving normally. Thus the
challenge for Rochefort and Michelakis (2) is to devise an
experiment that will demonstrate oxygen-sensitive ion chan-
nels in an intact pulmonary artery. Numerous studies of pul-
monary artery rings in vitro (provided they have experimental
verification that the endothelium was destroyed) have shown
that the hypoxic contraction is dependent on the presence of the
endothelium (6). However a sine qua non in these experiments
is the presence of “resting” arterial tone, a dynamic balance of
preconstruction and endogenous (mainly endothelial) vasodi-
lation. This is demonstrated by the dependence of hypoxic
contraction on the level of preconstriction and further contrac-
tion caused by inhibition of NO synthase (6). Under these
conditions, a contraction is caused by withdrawal of a tonic
vasodilator influence. Indeed inhibition of NO synthase pre-
vents hypoxic constriction of isolated artery rings (3, 4). Thus
the challenge for Robertson (5) is to devise an experiment that
will demonstrate that pulmonary rings generate a vasoconstric-
tor (that can be distinguished from loss of a vasodilator) to
match the time course of the contraction. It has already been
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shown that hypoxia inhibits NO formation by the endothelium
(1). NO regulates arterial tone through simultaneous modula-
tion of several pathways, one of which is to inhibit calcium
sensitization. Thus the evidence for an involvement of Rho or
Rho kinase in HPV is compatible with NO withdrawal.
REFERENCES
1. Demiryürek AT, Wadsworth RM, Kane AJ, Peacock KA. The role of
endothelium in hypoxic constriction of human pulmonary artery rings. Am
Rev Resp Dis 147: 283–290, 1993.
2. Rochefort GY, Michelakis ED. Counterpoint: Release of an endothelium-
derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitiza-
tion of smooth muscle cell contraction are not the main effectors for full and
sustained pulmonary vasoconstriction. J Appl Physiol. In press.
3. Karamsetty MR, Wadsworth RM, Kane KA. Effects of K⫹ channel
blocking drugs and nitric oxide synthase inhibition on the response to
hypoxia in rat pulmonary artery rings. J Auton Pharmacol 18: 49 –56, 1998.
4. Karamsetty MR, MacLean MR, McCulloch KM, Kane KA, Wads-
worth RM. Hypoxic constrictor response in the isolated pulmonary artery
from chronically hypoxic rats. Respir Physiol 105: 85–93, 1996.
5. Robertson TP. Point: Release of an endothelium-derived vasoconstrictor
and RhoA/Rho kinase-mediated calcium sensitization of smooth muscle
cell contraction are the main effectors for full and sustained pulmonary
vasoconstriction. J Appl Physiol. In press.
6. Wadsworth RM. Vasoconstrictor and vasodilator effects of hypoxia.
Trends Pharmacol Sci 15: 47–53, 1994.
Roger M. Wadsworth
University of Strathclyde
Glasgow, United Kingdom
To the Editor: We very much enjoyed the authors’ elegant
papers and we agree with the conclusion that Rho kinase plays
an important role in the sustained phase of hypoxic pulmonary
vasoconstriction (4). In regard to the endothelium-derived
vasoconstrictor, selective endothelin receptor A blockade has
been shown to attenuate acute and chronic HPV in both intact
rats and isolated rat lungs (5), suggesting that the endothelin
system plays a significant role in HPV. However, as Drs.
Rochefort and Michelakis (1) point out, it seems unlikely that
a process as complex as HPV should have a single dominant
culprit mediator. It appears plausible that sustained HPV,
similar to other processes such as the regulation of systemic
vascular resistance, is mediated by multiple mechanisms. For
2077
Point:Counterpoint Comments
2078
example, sustained HPV is also mediated by protein kinase C
and p38 MAP kinase (2, 6). Voltage-gated K⫹ channels have
been proposed to be a central mediator of HPV since their
regional selectivity to the pulmonary vasculature matches that
of HPV. However, while both restoration of K⫹ channel
function and Rho kinase inhibition decrease pulmonary artery
pressures in chronic hypoxic vasoconstriction, neither ap-
proach has been shown to completely normalize PA pressures
(3). We have learned from several diseases that therapeutic
manipulation of multiple pathways is more beneficial than
treating one single pathophysiological mechanism alone. In
addition, many investigators are also performing the needed
examinations of pharmacogenetics and sex differences in these
models. To attenuate the deleterious effects of sustained HPV
(like chronic pulmonary hypertension and cor pulmonale), a
multimodal approach targeting Rho kinase and endothelial
factors as well as redox balance and voltage-gated K⫹ channels
will hopefully drive us close to perfect control.
REFERENCES
1. Michelakis E, Rochefort G. Counterpoint: Release of an endothelium-
derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitiza-
tion of smooth muscle cell contraction are not the main effectors for full and
sustained hypoxic pulmonary vasoconstriction. J Appl Physiol. In press.
2. Morrell ED, Tsai BM, Wang M, Crisostomo PR, Meldrum DR. P38
mitogen-activated protein kinase mediates the sustained phase of hypoxic
pulmonary vasoconstriction and plays a role in phase I vasodilation. J Surg
Res 134: 335–341, 2006.
3. Morrell ED, Tsai BM, Crisostomo PR, Wang M, Markel TA, Lillemoe
KD, Meldrum DR. Therapeutic concepts for hypoxic pulmonary vasocon-
striction involving ion regulation and the smooth muscle contractile appa-
ratus. J Mol Cell Cardiol 40: 751–760, 2006.
4. Robertson T. Point: Release of an endothelium-derived vasoconstrictor
and RhoA/Rho kinase-mediated calcium sensitization of smooth muscle
cell contraction are the main effectors for full and sustained hypoxic
pulmonary vasoconstriction. J Appl Physiol. In press.
5. Sato K, Morio Y, Morris KG, Rodman DM, McMurtry IF. Mechanism
of hypoxic pulmonary vasoconstriction involves ETA receptor-mediated
inhibition of KATP channel. Am J Physiol Lung Cell Mol Physiol 278:
L434 –L442, 2000.
6. Tsai BM, Wang M, Pitcher JM, Meldrum KK, Meldrum DR. Hypoxic
pulmonary vasoconstriction and pulmonary artery tissue cytokine expres-
sion are mediated by protein kinase C. Am J Physiol Lung Cell Mol Physiol
287: L1215–L1219, 2004.
Tim Lahm
Daniel R. Meldrum
Indiana University
School of Medicine
Indianapolis, Indiana
To the Editor: I appreciate the frank exchange of views and
comprehensive discussion between both authors and enjoy
reading the discussions on the issue (1, 3). The major function
of the lung is gas exchange, and hypoxic pulmonary vasocon-
striction (HPV) is undoubtedly a critical physiological mech-
anism for maximizing oxygenation of the venous blood. An
important physiological mechanism like HPV must have mul-
tiple pathways to assure its effectiveness and consistency. To
ensure the reliability of HPV, multiple cell types, different
mediators, various membrane proteins, and diverse intracellu-
lar signaling cascades should be involved (and accurately
coordinated) in stimulating smooth muscle to contract (and to
relax). A rise in cytosolic Ca2⫹ ([Ca2⫹]cyt), along with an
increase in Ca2⫹ sensitization, has been implicated as a major
J Appl Physiol • VOL
mechanism involved in smooth muscle contraction or vasocon-
striction. How many proteins, molecules, and signaling path-
ways are involved in regulating [Ca2⫹]cyt, binding Ca2⫹/cal-
modulin to contractile proteins, or movement of actomyosin?
The answer is, a lot. Therefore, why do we fantasize for a
single mechanism, or the mechanism, by which alveolar hyp-
oxia causes pulmonary vasoconstriction? Can’t we trust the
outcome of evolution that may have precisely “designed” a
heterogeneous network to guarantee HPV? Isn’t it possible
that alveolar hypoxia, the initial trigger for HPV, is just a
“playboy” who uses whatever he can get (simply, easily,
quickly, and inexpensively) to achieve the goal? As an old
saying goes, all roads lead to Rome. It is time for all inves-
tigators to sit together in a meeting held at a high-altitude or
hypoxic environment and try to draw a map of the network
(2, 4 – 6).
REFERENCES
1. Michelakis E, Rochefort G. Counterpoint: Release of an endothelium-
derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitiza-
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tion of smooth muscle cell contraction are not the main effectors for full and
sustained hypoxic pulmonary vasoconstriction. J Appl Physiol. In press.
2. Moudgil R, Michelakis ED, Archer SL. Hypoxic pulmonary vasocon-
striction. J Appl Physiol 98: 390 – 403, 2005.
3. Robertson T. Point: Release of an endothelium-derived vasoconstrictor
and Rhoa/Rho kinase-mediated calcium sensitization of smooth muscle cell
contraction are the main effectors for full and sustained hypoxic pulmonary
vasoconstriction. J Appl Physiol. In press.
4. Ward JPT, Aaronson PI. Mechanisms of hypoxic pulmonary vasocon-
striction: can anyone be right? Respir Physiol 115: 261–267, 1999.
5. Weir EK, Barneo JL, Buckler KJ, Archer SL. Acute oxygen-sensing
mechanisms. N Engl J Med 353: 2042–2055, 2005.
6. Yuan JX-J. (editor). Hypoxic Pulmonary Vasoconstriction: Cellular and
Molecular Mechanisms. Boston, MA: Kluwer Academic, 2004.
Jason X-J. Yuan
University of California
San Diego, California
To the Editor: Whether hypoxia causes vasoconstriction or
vasodilation is dependent on vascular smooth muscle type and
not on presence of the endothelium (5, 6). Mammalian pulmo-
nary arterial and venous muscles both contract to hypoxia (5,
6), a phenomenon defined as hypoxic pulmonary vasoconstric-
tion (HPV). Yet arteries and veins have dissimilar endothelia.
Hypoxic vasoconstriction is not limited to mammalian pulmo-
nary vasculature but is manifested in a variety of vessels from
a variety of species (1, 4). While Robertson argues that HPV is
often biphasic (2), biphasic responses are actually reported in
relatively few vessel types from relatively few species, perhaps
only in rat pulmonary artery. Rat pulmonary veins respond to
hypoxia with a monophasic contraction (5). Robertson refers
only to literature that supports the idea that the second sus-
tained phase of rat HPV is endothelium dependent (2). How-
ever, others report that the early short-lived phase of hypoxic
contraction is the only endothelium-dependent component (6).
Importantly, Rochefort and Michelakis (3) point out that nei-
ther the biphasic nature nor the pretone or pretreatment require-
ment (i.e., preconditioning or precontracting) of the rat pulmo-
nary arterial response to hypoxia is a universal feature of HPV.
Indeed, pig, hagfish, lamprey, and the California blackworm
can be added to the list of species in which hypoxic vasocon-
striction occurs without preconditioning (1, 4). Perhaps the
102 • MAY 2007 • www.jap.org

conflicting results of studies in which the endothelium has been
removed might be explained by trauma to the underlying
smooth muscle that occurs in the hands of some investigators
but not in the hands of others during the endothelial denudation
process.
REFERENCES
1. Phelps S, Pelaez NJ. Hypoxia narrows and prolongs contraction of the
dorsal blood vessel in the California blackworm, Lumbriculus variegatus.
FASEB J 17: A425, 2003.
2. Robertson T. Point: Release of an endothelium-derived vasoconstrictor
and RhoA/Rho kinase-mediated calcium sensitization of smooth muscle
cell contraction are the main effectors for full and sustained hypoxic
pulmonary vasoconstriction. J Appl Physiol. In press.
3. Rochefort GY, Michelakis ED. Counterpoint: Release of an endothelium-
derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitiza-
J Appl Physiol • VOL 102 • MAY 2007 •
Point:Counterpoint Comments
2079
tion of smooth muscle cell contraction are not the main effectors for full and
sustained hypoxic pulmonary vasoconstriction. J Appl Physiol. In press.
4. Russell MJ, Pelaez NJ, Packer CS, Forster ME, Olson KE. Intracellular
and extracellular calcium utilization during hypoxic vasoconstriction in
cyclostome aortas. Am J Physiol Regul Integr Comp Physiol 281: R1506 –
R1513, 2001.
5. Zhao Y, Packer CS, Rhoades RA. Response of rat isolated pulmonary
vein to hypoxia. Am J Physiol Lung Cell Mol Physiol 265: L89 –L92, 1993.
6. Zhao Y, Rhoades RA, Packer CS. Hypoxia induced pulmonary arterial
contraction appears to be dependent on myosin light chain phosphorylation.
Am J Physiol Lung Cell Mol Physiol 271: L768 –L774, 1996.
C. Subah Packer1
Nancy J. Pelaez2
1
Cellular and Integrative Physiology
Indiana University School of Medicine
2
National Science Foundation
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Letters To The Editor
Last Word on Point:Counterpoint “Release of an endothelium-derived
vasoconstrictor and RhoA/Rho kinase-mediated calcium sensitization of
smooth muscle cell contraction are/are not the main effectors for full and
sustained HPV”
T. P. Robertson
University of Georgia, Athens, Georgia
To the Editor: I would like to congratulate Drs. Rochefort and
Michelakis (3) for a most enjoyable and spirited debate!
Drs. Madden and Packer (2) both intimate that one possible
explanation for the loss of HPV following endothelial denuda-
tion could be that some experimentalists damage the underly-
ing smooth muscle during the denudation process. However, it
has been reported several times that denudation eliminates
sustained HPV, whereas smooth muscle-specific contractile
responses, such as potassium-induced depolarization, are un-
affected (e.g., Ref. 4).
Indeed, when considering potential “damage” to smooth mus-
cle and the resultant effects on function, I would concur with Dr.
Wadsworth (2) when posing the question as to whether enzymat-
ically dispersed cells in culture are behaving normally?
Dr. Wadsworth makes a case for sustained HPV being the
result of an inhibition of the release of endothelium-derived
nitric oxide by hypoxia. As with many aspects of HPV, there
is evidence both for and against this hypothesis. However, the
balance of data seems to tip against this possibility, and it is
more probable that NO may be increased in HPV rather than
decreased (discussed in Ref. 1).
Address for reprint requests and other correspondence: T. P. Robertson,
Univ. of Georgia, Athens, GA 30605 (e-mail: troberts@vet.uga.edu).
2080 8750-7587/07 $8.00 Copyright © 2007 the American Physiological Society
J Appl Physiol 102: 2080, 2007;
doi:10.1152/japplphysiol.00002.2007.
Drs. Meldrum, Lahn, and Yuan (2) seek to bring an accord
to the current debate by suggesting that HPV is comprised of
a concert of inter- and intradependent signal transduction
pathways acting in a harmonious fashion to elicit the desired
contractile response. At this point, we must put this notion in
the context of the Point:Counterpoint debate, which is to avoid
objectivity and be completely subjective. Objectively, I concur
with Drs. Meldrum, Lahn, and Yuan and agree that HPV is
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most certainly a well orchestrated multi-factorial process. Sub-
jectively, it’s the endothelium, dummy!
REFERENCES
1. Aaronson PI, Robertson TP, Ward JPT. Endothelium-derived mediators
and hypoxic pulmonary vasoconstriction. Resp Physiol Neurobiol 132:
107–120, 2002.
2. Madden JA, Wadsworth RM, Meldrum DR, Lahm T, Yuan J, Packer
CS, Pelaez NJ. Comments on Point:Counterpoint: Release of an endothe-
lium-derived vasoconstrictor and RhoA/Rho kinase-mediated calcium sen-
sitization of smooth muscle cell contraction are/are not the main effectors
for full and sustained HPV. J Appl Physiol. In press.
3. Rochefort GY, Michelakis ED. Counterpoint: Release of an endothelium-
derived vasoconstrictor and Rhoa/Rho kinase-mediated calcium sensitiza-
tion of smooth muscle cell contraction are not the main effectors for full and
sustained HPV. J Appl Physiol. In press.
4. Robertson TP, Aaronson PI, Ward JPT. Ca2⫹ sensitization during
sustained hypoxic pulmonary vasoconstriction is endothelium dependent.
Am J Physiol Lung Cell Mol Physiol 284: L1121–L1126, 2003.
http://www. jap.org