PHYSIOLOGY, ENDOCRINOLOGY, AND REPRODUCTION
Analysis of Plasma Serotonin Levels and Hemodynamic Responses
Following Chronic Serotonin Infusion in Broilers Challenged
with Bacterial Lipopolysaccharide and Microparticles1
M. E. Chapman,*2 R. L. Taylor,† and R F. Wideman Jr.*
*Department of Poultry Science, University of Arkansas, Fayetteville 72701;
and †Department of Animal and Nutritional Sciences, University of New Hampshire, Durham 03824
ABSTRACT There has been extensive interest in the
role of serotonin (5-hydoxytryptamine, 5-HT) in the
pathogenesis of pulmonary hypertension because epi-
sodes of pulmonary arterial hypertension in humans have
been linked to serotoninergic appetite-suppressant drugs.
In this study, we investigated the role of serotonin in
the development of pulmonary hypertension induced by
intravenously injecting bacterial lipopolysaccharide (LPS,
endotoxin) and cellulose microparticles. In experiment 1,
we used a 5-HT ELISA kit for the in vitro quantitative
determination of 5-HT in plasma during the development
of pulmonary hypertension induced by injecting LPS and
cellulose microparticles i.v. in broilers. In experiment 2,
broilers were either chronically infused with 5-HT via
surgically implanted osmotic pumps or received sham
surgery as a control. After a period of 10 d, the pulmonary
arterial pressure was recorded during challenge with in-
jected LPS or microparticles. Microparticles elicited 5-HT
plasma levels more than 2-fold greater than those elicited
by LPS from 15 to 45 min postinjection. This indicates
Key words: hypertension, broiler, serotonin, lipopolysaccharide, microparticle
INTRODUCTION
Fast-growing broilers are susceptible to pulmonary ar-
terial hypertension (PAH), leading to pulmonary hyper-
tension syndrome (ascites) when their right ventricle
must progressively elevate the pulmonary arterial pres-
sure (PAP) to propel the requisite cardiac output through
lungs having a marginally inadequate pulmonary vascu-
©2008 Poultry Science Association Inc.
Received April 18, 2007.
Accepted September 10, 2007.
1
United States patent no. 6,720,473 protects the exclusive rights of
the University of Arkansas to all uses of the intravenous micro-particle
injection technology within the context of evaluating or affecting pulmo-
nary vascular capacity, pulmonary vascular resistance, pulmonary hy-
pertension, cardio-pulmonary hemodynamics, and susceptibility to pul-
monary hypertension and pulmonary hypertension syndrome (ascites)
in domesticated animal species.
2
Corresponding author: mchapman@uark.edu
116
that 5-HT is an important mediator in the pulmonary
hypertensive response of broilers to microparticles, but
may not play a prominent role in the pulmonary hyper-
tensive response to LPS. Furthermore, chronic 5-HT infu-
sion via osmotic pumps caused an increase in the duration
of the pulmonary hypertensive response of broilers to
microparticles, indicating that the infused 5-HT was se-
questered by circulating thrombocytes and then released
upon microparticle-mediated thrombocyte activation. Se-
rotonin appears to play a less prominent role in the pul-
monary hypertensive response of broilers to LPS, indicat-
ing that other mediators within the innate response to
inflammatory stimuli may also be involved. These results
are consistent with our hypothesis that pulmonary arte-
rial hypertension ensues when vasoconstrictors such as
5-HT overwhelm the dilatory affects of vasodilators such
as nitric oxide, thereby effectively reducing the pulmo-
nary vascular capacity of pulmonary arterial hyperten-
sion-susceptible broilers.
2008 Poultry Science 87:116–124
doi:10.3382/ps.2007-00160
lar capacity (Peacock et al., 1989; Julian, 1993; Wideman
and Bottje, 1993; Wideman, 2000, Wideman et al., 2001).
However, we still do not know what initiates PAH. Vaso-
constriction, remodeling of the pulmonary vessel wall,
and thrombosis are all thought to contribute to increased
pulmonary vascular resistance in PAH. Any factor that
increases the cardiac output, reduces the pulmonary vas-
cular capacity, or triggers pulmonary vasoconstriction
can contribute to the pathogenesis of PAH in broilers
(Wideman, 2000), particularly when occurring within a
genetic predisposition. Indeed, several cell types, includ-
ing endothelial and smooth muscle cells, as well as in-
flammatory cells and thrombocytes, may play a signifi-
cant role in PAH. There has been extensive interest in
the role of serotonin (5-hydoxytrytamine, 5-HT) in the
pathogenesis of pulmonary hypertension, because epi-
sodes of PAH in humans have been linked to serotoniner-
gic appetite-suppressant drugs (Abenhaim et al., 1996).
Increased plasma 5-HT levels have also been recorded in
 ROLE OF SERATONIN IN PULMONARY ARTERIAL HYPERTENSION
human patients who spontaneously developed idiopathic
PAH (Hervé et al., 1995).
Serotonin is a potent pulmonary vasoconstrictor (Boe
et al., 1980) and smooth muscle mitogen (Nemecek et al.,
1986; Fanburg and Lee, 1997) that is synthesized from the
essential amino acid Trp. It is actively accumulated by
mammalian platelets and avian thrombocytes, where 98%
or more of all circulating 5-HT is stored, and is released
into the plasma during platelet or thrombocyte aggrega-
tion (Meyer and Sturkie, 1974; Cox, 1985; Lacoste-
Eleaume et al., 1994). Serotonin is taken up by the 5-HT
transporter (5-HTT) in several cells, including platelets
and thrombocytes, as well as in pulmonary artery smooth
muscle and endothelial cells. Serotonin has been impli-
cated in the mechanisms responsible for pulmonary hy-
pertension in several human, animal, and broiler studies
(Douglas et al., 1981; Abenhaim et al., 1996; Chapman
and Wideman, 2002). When platelets and thrombocytes
aggregate, they release several physiologically active sub-
stances, including 5-HT, which causes proliferation of
pulmonary vascular smooth muscle cells and stimulates
vasoconstriction, thereby reducing the blood flow at the
site of injury (Lee et al., 1994; Fanburg and Lee, 1997).
Pulmonary vasoconstriction induced by 5-HT is believed
to be mediated through 5-HT1B/1D and 5-HT2A receptors
expressed by pulmonary smooth muscle cells, whereas
the vascular remodeling associated with pulmonary hy-
pertension appears to require the serotonin transporter
(5-HTT). Therefore, determination of altered 5-HT plasma
concentrations has great clinical significance for the as-
sessment of broiler PAH.
In experiment 1, we used an enzyme immunoassay for
the in vitro quantitative determination of plasma 5-HT
during the development of pulmonary hypertension over
a 12-h period, induced by injecting bacterial lipopolysac-
charide (LPS) or cellulose microparticles i.v. in broilers.
Lipopolysaccharide and microparticles cause pulmonary
vasoconstriction and pulmonary hypertension in broilers
(Wideman et al., 2001; Wideman and Erf, 2002), but the
specific mediator of vasoconstriction has not been deter-
mined. Bacterial LPS can produce direct injury to the
pulmonary vascular endothelium (Brigham and Meyrick,
1986; Meyrick et al., 1986); however, the pulmonary hy-
pertension in gram-negative sepsis primarily results from
LPS-activated host inflammatory responses (Brigham and
Meyrick, 1986; Ghosh et al., 1993). Entrapped micropar-
ticles, in addition to physically occluding pulmonary arte-
rioles, stimulate local tissues and leukocytes to release
vasoactive substances capable of altering pulmonary vas-
cular resistance by dilating or constricting the nearby
vasculature (Wang et al., 2003; Wideman et al., 2004).
Therefore, the objective of experiment 1 was to determine
whether LPS- or microparticle-induced pulmonary hy-
pertensive responses in broilers corresponded to in-
creased plasma 5-HT levels. In experiment 2, broilers
were either chronically infused with 5-HT via surgically
implanted osmotic pumps or received sham surgery as
a control (McCorkle and Taylor, 1989; Eddahibi et al.,
1997). After a period of 10 d, the broilers were injected
117
with LPS or microparticles while recording PAP. Experi-
ment 2 had the objective of determining whether supple-
mental 5-HT infusions would exacerbate the pulmonary
hypertensive response of broilers to LPS or micropar-
ticles.
MATERIALS AND METHODS
Male broilers from a commercial hatchery (Cobb-Van-
tress Inc., Fayetteville, AR) were wing-banded and trans-
ported on the day of hatch (d 1) to the Poultry Environ-
mental Research Laboratory at the University of Arkan-
sas. They were placed on fresh wood shavings in
environmental chambers (8 m2 of floor space) and were
brooded at 33°C from d 1 to 5, 29°C from d 6 to 10,
and 27°C from d 11 to 17. Thereafter, the broilers were
maintained at 23.8°C until the experiment was termi-
nated. The photoperiod was 24 h of light from d 1 to 5,
and 23L:1D thereafter. Water was provided ad libitum
via nipple type waterers. A corn-soybean meal starter
ration (22.7% CP, 3,059 kcal of ME/kg, 1.5% Arg, and
1.43% Lys) was provided ad libitum and was formulated
to meet minimum NRC (1994) standards for all ingredi-
ents. The diet was provided as crumbles during wk 1 and
2 and as pellets thereafter.
Experiment 1
From d 31 to 36, unanesthetized male broilers (n = 240;
1,904 ± 15.6 g of BW, mean ± SEM) were injected via the
basilica vein with either 1 mg of Salmonella enterica serovar
Typhimurium LPS (Sigma Chemical Co., St. Louis, MO;
0.5 mL dissolved at 2 mg/mL in a 0.9% sodium chloride
solution, n = 120) or 0.35 mL of microgranular CM-32
ion-exchange cellulose microparticles (Fisher Scientific,
St. Louis, MO), suspended at 0.02 g/mL in heparinized
saline (n = 120), by using a 1-mL tuberculin syringe with
a 22-gauge needle. Previous studies demonstrated that
the l-mg dose of LPS elicits a maximal pulmonary hyper-
tensive response in broilers (Wideman et al., 2001) with-
out triggering endotoxin shock (Xie et al., 2000) and that
approximately 0.3 to 0.35 mL of the 0.02 g/mL micropar-
ticle suspension is required to elevate the PAP of 6- to 7-
wk-old male broilers significantly while triggering <10%
mortality within 24 h postinjection (Wideman and Erf,
2002; Wideman et al., 2005b). At 15, 30, and 45 min, and
1, 2, 3, 4, 5, 6, 8, 10, and 12 h postinjection, 1.5-mL blood
samples were collected from the basilica vein by using
syringes containing 0.1 mL of a 0.9% sodium chloride
solution containing 50 mg/mL of EDTA. The EDTA was
used to chelate calcium and thereby prevent platelet acti-
vation and degranulation (Cox, 1985; Gobbi et al., 2003).
The plasma was separated by centrifugation and was
stored frozen at −4°C until analysis. Plasma 5-HT levels
were measured by using a commercial enzyme immuno-
assay kit (Alpco Diagnostics, Windham, NH).
Experiment 2
Beginning on d 34 and continuing at daily intervals
thereafter during the ensuing week, unanesthetized male
118 CHAPMAN ET AL.
broilers (n = 40; 2,307 ± 139.1 g of BW, mean ± SEM)
were placed on a surgical board and restrained in dorsal
recumbency. The feathers were removed from the dorsal
surface of the neck as needed, intracutaneous injections
of 2% lidocaine HCl were administered as a local anesthe-
tic, and an incision was made adjacent to the site chosen
for the subcutaneous surgical implantations. Osmotic
pumps (2ML2 Alzet, Durect Corporation, Cupertino, CA)
containing 2 mL of 5-HT (Sigma Chemical Co.) at 10 mg/
mL in 0.9% saline (pump group, n = 20) or gelatin capsules
(sham group, n = 20) were implanted and the implanta-
tion site was then closed with wound clips. The 2ML2
Alzet osmotic pumps are designed to deliver a test drug
at the consistent rate of 5 ␮L/h for 14 d. Based on a
previous study (Chapman and Wideman, 2002) the 10
mg/mL concentration of 5-HT was chosen to increase 5-
HT levels in circulating thrombocytes without causing
the pulmonary vasoconstriction leading to PAH.
Ten to 12 d postimplantation (starting on d 44), the
broilers were anesthetized to a light surgical plane with
intramuscular injections of allobarbitol (5,5-diallylbarbi-
turic acid, 3.0 mL, 25 mg/mL, Sigma Chemical Co.) and
ketamine HCl (1.0 mL, 100 mg/mL, Bedford Laboratories,
Bedford, OH). The birds were placed on a heated surgical
board (30°C) and restrained in dorsal recumbency. The
left wing was extended and feathers were removed from
the ventral surface as needed to uncover the skin over
the basilica vein. After intracutaneous injections of 2%
lidocaine HCl were administered as a local anesthetic, an
incision was made to expose the vein, which then was
cannulated with a Silastic catheter (0.03 cm i.d., 0.09 cm
o.d.) filled with a 0.9% sodium chloride solution con-
taining 200 IU of heparin/mL. The catheter was attached
to a blood pressure transducer interfaced through a
Transbridge preamplifier (World Precision Instruments,
Sarasota, FL) to a Biopac MP 100 data acquisition system
using AcqKnowledge software (Biopac Systems Inc., Go-
leta, CA). The catheter was advanced through the right
atrium and ventricle into a pulmonary artery while the
characteristic pulse pressures were monitored to identify
the location (Wideman et al., 1996; Chapman and Wide-
man, 2001). All pulmonary arterial pressure readings
were made with the transducer at the level of the thoracic
inlet. Prior to recording, the system was calibrated in
millimeters of mercury (mmHg) by using a mercury ma-
nometer. The left basilica vein was also cannulated with
PE-50 polyethylene tubing filled with heparinized saline
for i.v. injections. Control data were recorded for 10 min
after surgical preparations were complete and a stabiliza-
tion period of 10 min had elapsed. The osmotic pump
and sham groups were then injected i.v. with 1 mg of
Salmonella Typhimurium LPS (0.5 mL dissolved at 2 mg/
mL in a 0.9% sodium chloride solution, n = 20) or 0.35
mL of cellulose microparticles suspended at 0.02 g/mL
in heparinized saline (n = 20). Data were recorded for a
further 60 min, after which the birds were euthanized
with 10 mL i.v. of 0.1 M KCl.
Data Analysis
The primary channel of the Biopac MP 100 data acquisi-
tion system recorded PAP (mmHg). These data was aver-
aged electronically during representative sample inter-
vals while accommodating for the influences of pulse
pressure and respiratory cycles on pulmonary arterial
pressure (Wideman et al., 1996). Data were analyzed by t-
test (comparison between 2 groups within a single sample
interval) or within a group over time (across sample inter-
vals) by using the SigmaStat Repeated Measures ANOVA
procedure (Jandel Scientific, 1994). The Dunnett method
was used for the separation of treatment means. The
threshold for significance in all cases was P ≤ 0.05.
RESULTS
Experiment 1
At 15, 30, and 45 min postinjection, microparticles elic-
ited plasma levels of 5-HT of 171 ± 40, 234 ± 80, and 175
± 50 ng/mL, respectively during the pulmonary arterial
hypertensive response in broilers (Figure 1). This 5-HT
response to microparticles was more than 2-fold greater
(P = 0.04) than the responses elicited by LPS (69 ± 20, 31
± 13, and 80 ± 50 ng/mL, respectively). With the exception
of a significant difference at the 6-h sample interval, the
plasma 5-HT levels of broilers injected with micropar-
ticles were similar to 5-HT levels observed in broilers
injected with LPS. The 5-HT concentrations in micropar-
ticle-injected birds declined after the 30-min mea-
surement.
Experiment 2
For broilers that subsequently were injected with LPS,
the initial baseline PAP averaged approximately 20
mmHg in the broilers from the sham group, whereas after
10 d of chronic 5-HT infusion via osmotic pumps, the
baseline PAP averaged approximately 23 mmHg in the
pump group (sample intervals St to S10; Figure 2a). The
numerically greater initial baseline PAP values for broil-
ers in the pump group apparently shifted the group’s
absolute PAP response to LPS upward by approximately
3 mmHg when compared with the absolute PAP response
to LPS by the sham group (sample intervals T to T60).
Accordingly, absolute PAP values for both groups were
recalculated to reflect the LPS-induced percentage change
from the initial baseline PAP values, as shown in Figure
2b. Injecting 1 mg of LPS into the wing vein caused the
PAP to rise within 10 min and reach a peak of approxi-
mately 70% above baseline values within 25 min (P >
0.05) in both groups of broilers, regardless of treatment
during the preceding 10 d (5-HT infusion, sham controls).
The percentage change in PAP then subsided toward
baseline values by the end of the experiment in both the
5-HT-treated and control groups. At no sample interval
during the hypertensive response to LPS (T to T60) did
the percentage change in PAP differ between the groups
(P > 0.05).
 ROLE OF SERATONIN IN PULMONARY ARTERIAL HYPERTENSION 119

Figure 1. Plasma serotonin levels (ng/mL) for male broilers (n = 240; 1,904 ± 15.6 g of BW, mean ± SEM) in experiment 1, at 15, 30, and 45 min
and at 1, 2, 3, 4, 5, 6, 8, 10, and 12 h after lipopolysaccharide (LPS) or microparticle injection. Subscript letters (a and b) represent differences (P
≤ 0.05) between groups within individual sample intervals. Asterisks (*) represent differences (P ≤ 0.05) within a group across sample intervals.
5-HT = 5-hydroxytryptamine (serotonin).

For broilers that were subsequently injected with mi-
croparticles, the initial baseline PAP was approximately
16.5 mmHg in birds from the sham group and approxi-
mately 18.5 mmHg in birds that had received chronic 5-
HT infusion via osmotic pumps (sample intervals St to
S10; Figure 3a). Because of the numerically greater initial
absolute PAP values in broilers from the pump group
when compared with the sham broilers, the percentage
change from the initial PAP was calculated as shown
in Figure 3b for comparisons of the pulmonary arterial
hypertensive response between the groups. Injecting mi-
croparticles into the wing vein caused the PAP to rise
within 5 min to a peak of approximately 50% above base-
line values (P > 0.05) in broilers, regardless of treatment
(sample interval TPk, sham and pump groups). The per-
centage change in PAP remained elevated by approxi-
mately 20% above baseline values by the end of the experi-
ment (sample interval T60) in broilers that were chroni-
cally infused with 5-HT, whereas in broilers from the
sham group, the pulmonary hypertensive response sub-
sided 20 min earlier (sample interval T40) to a level that
did not differ from the initial baseline values. At no sam-
ple interval during the hypertensive response to micro-
particles (T to T60) did the absolute PAP or percentage
change in PAP differ between the groups (P > 0.05).
DISCUSSION
Previously, we demonstrated that the nonselective 5-
HT1/2 receptor antagonist methiothepin virtually elimi-
nated the pulmonary hypertensive response of broilers
to i.v. microparticle injections but had minimal impact on
LPS-mediated pulmonary hypertension (Chapman and
Wideman, 2006b). However, changes in plasma 5-HT lev-
els following LPS or microparticle injections had not pre-
viously been demonstrated. In experiment 1 at 15 to 45
min postinjection, the microparticles elicited plasma lev-
els of 5-HT more than 2-fold greater than those elicited
by LPS in broilers. Plasma 5-HT levels then declined over
the remainder of the experiment toward levels similar to
those observed in broilers injected with LPS. The present
and previous studies have consistently indicated that 5-
HT is an important mediator of broilers’ pulmonary hy-
pertensive response to microparticles; however, 5-HT
may not play a prominent role in the pulmonary hyper-
tensive response of broilers to LPS. This suggests that
other mediators of the innate immune response interact
with LPS.
It has been shown that within minutes after being in-
jected, entrapped microparticles are surrounded by focal
aggregates of thrombocytes and monocytes or macro-
phages (Wideman et al., 2002; Wang et al., 2003), which
potentially can trigger these leukocytes and the adjacent
vascular endothelium to synthesize and release potent
vasoactive compounds within close proximity to the vas-
cular smooth muscle (Wideman, 2001; Wideman et al.,
2004). The tendency for elevated plasma 5-HT levels to
wane within 1 h postmicroparticle injection, despite the
persistence of cellulose microparticles within the pulmo-
120 CHAPMAN ET AL.

Figure 2. a) Pulmonary arterial pressure (PAP) and b) percentage change from the initial PAP (PAP % change) for male broilers (n = 40; 2,307
± 139.1 g of BW, mean ± SEM) in experiment 2, at the start of data collection and at intervals of 5 min thereafter (St, S5, S10), within 30 s after
lipopolysaccharide (LPS) injection and at intervals of 5 min thereafter (T to T60). Asterisks (*) represent differences (P ≤ 0.05) within a group across
sample intervals.

nary microvasculature for more than 10 d (Wideman et
al., 2002; Wang et al., 2003), may be attributable to exhaus-
tion of thrombocyte 5-HT reserves, cessation of thrombo-
cyte activation, or inhibition by nitric oxide (NO; Moro
et al., 1995). The available 5-HT also may be internalized
into pulmonary endothelial cells via the 5-HTT. Pulmo-
nary arterial hypertension is characterized by vasocon-
striction, followed by structural remodeling associated
with smooth muscle cell proliferation, both of which can
be caused by increases in plasma 5-HT. Pulmonary vaso-
constriction is thought to be mediated by 5-HT2A and 5-
HT1B/1D receptors expressed on vascular smooth muscle
cells (Choi and Maroteaux, 1996; MacLean et al., 1996;
Keegan et al., 2001). Structural remodeling is mediated
by 5-HTT expressed on vascular smooth muscle cells in
the lungs. Serotonin translocated into the smooth muscle
 ROLE OF SERATONIN IN PULMONARY ARTERIAL HYPERTENSION 121

Figure 3. a) Pulmonary arterial pressure (PAP) and b) percentage change from the initial PAP (PAP % change) for male broilers (n = 40; 2,307
± 139.1 g of BW, mean ± SEM) in experiment 2, at the start of data collection and at intervals of 5 min thereafter (St, S5, S10), within 30 s after
microparticle injection and at intervals of 5 min thereafter (T to T60). Asterisks (*) represent differences (P ≤ 0.05) within a group across sample intervals.

cells by 5-HTT is mitogenic, causing hypertrophy and
proliferation of the medial muscle layer in pulmonary
arterioles (Lee et al., 1991, 1994; Ramamoorthy et al., 1993;
Fanburg and Lee, 1997; Eddahibi et al., 1999).
Serotonin, it seems, may not play a prominent role in
the pulmonary hypertensive response to LPS in broilers,
suggesting that the pathological progression toward PAH
has more than one progenitor. The vasoconstrictor throm-
boxane A2 (TxA2) has been shown to increase pulmonary
vascular resistance and PAP in broilers (Wideman et al.,
1998a, 1999a, 2001), and several studies have associated
TxA2 with the pulmonary hypertension during endotoxe-
mia. Levels of thromboxane B2 (a stable metabolite of
TxA2) in plasma and lung lymph increased acutely after
LPS administration in several mammalian species (Ball
et al., 1983; Snapper et al., 1983; Kubo and Kobayashi,
122 CHAPMAN ET AL.
1985). Furthermore, the fawn-hooded rat, which sponta-
neously develops PAH, is believed to be a model of 5-
HT-mediated PAH owing to a platelet storage defect that
impairs uptake and retention of serotonin and elevates
plasma serotonin levels (Tschopp and Zucker, 1972; Gon-
zalez et al., 1998). However, recent studies have impli-
cated overexpression of endothelin-1 (ET-1) mRNA and
overproduction of the pulmonary vasoconstrictor ET-1
peptide in fawn-hooded rat lung tissues as the potential
mediator of PAH (Hahn et al., 1990; Nagaoka et al., 2001).
Additional studies using fawn-hooded rat lungs point
toward impaired endothelial NO synthase expression
(Tyler et al., 1999), leading to sustained pulmonary vaso-
constriction and PAH. Endothelial dysfunction has been
identified as playing a key role in PAH pathophysiology,
leading to impaired release of NO and prostacyclin and an
overexpression of vasoconstrictors such as thromboxane
and endothelin-1 (Humbert et al., 2004). Pulmonary hy-
pertension therefore appears to have a multifactorial pa-
thology.
In experiment 2, chronic (10-d) 5-HT infusion via os-
motic pumps caused a numerical increase in PAP in com-
parison with control broilers that had received sham sur-
gery, suggesting a role for 5-HT in maintaining the basal
tone of the pulmonary arterial wall in broilers. These
results are consistent with the observation that the 5-HT
receptor blocker methiothepin causes a 25% reduction in
baseline PAP in broilers (Chapman and Wideman,
2006a,b). The peak PAP response to 1 mg of LPS attained
a similar magnitude of 70% above baseline values within
25 min postinjection in both 5-HT-infused and sham
groups, providing further evidence that 5-HT may not
play a prominent role in the pulmonary hypertensive
response to LPS in broilers. Injecting microparticles into
the wing vein caused the PAP to rise within 5 min to a
peak of approximately 50% above baseline values in both
5-HT-infused and control broilers. Thereafter, the PAP
remained elevated by approximately 20% above baseline
values for the duration of the experiment in broilers
chronically infused with 5-HT but not in broilers from
the sham group. These observations are consistent with
the hypothesis that the infused 5-HT was sequestered by
circulating thrombocytes via the 5-HTT and then released
upon microparticle-mediated thrombocyte activation. En-
dothelial NO synthase can be induced in endothelial cells
to produce the vasodilator NO in response to increased
rates in blood flow through constricted vascular channels.
Previous studies have demonstrated that NO produced
by endothelial NO synthase likely contributes to the atten-
uation of the pulmonary hypertensive responses of broil-
ers to LPS and microparticles seen toward the end of
experiment 2 (Bowen et al., 2006; Wideman et al., 2006).
In summary, this study provides direct evidence that
5-HT plays an important role in the pulmonary hyperten-
sive response to intravenously injected microparticles in
broilers. Serotonin appears to play a less prominent role
in the pulmonary hypertensive response of broilers to
intravenously injected LPS, indicating that other media-
tors within the innate response to inflammatory stimuli,
such as TxA2, ET-1, or NO, may also be involved. The
current observations support our hypothesis that pulmo-
nary hypertension syndrome ensues when vasoconstric-
tors overwhelm the dilatory affects of vasodilators such
as NO.
ACKNOWLEDGMENTS
This project was supported by the National Research
Initiative of the USDA Cooperative State Research, Educa-
tion and Extension Service (Washington, DC), USDA/
CSREES/NRI grant no. 2003-35204-13392, and by an Ani-
mal Health Grant from the University of Arkansas Ag-
ricultural Experiment Station (Fayetteville, AR).
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