GASTROENTEROLOGY 2002;123:1120 –1128
The Role of the Gastric Afferent Vagal Nerve in Ghrelin-Induced
Feeding and Growth Hormone Secretion in Rats
YUKARI DATE,*,‡ NOBORU MURAKAMI,§ KOJI TOSHINAI,* SHIGERU MATSUKURA,* AKIRA NIIJIMA,㛳
HISAYUKI MATSUO,‡ KENJI KANGAWA,‡ and MASAMITSU NAKAZATO*
*Department of Internal Medicine, Miyazaki Medical College, Miyazaki; ‡National Cardiovascular Center Research Institute, Osaka;
§Department of Veterinary Physiology, Faculty of Agriculture, Miyazaki University, Miyazaki; and the 㛳Department of Physiology,
Niigata University School of Medicine, Niigata, Japan
Background & Aims: Visceral sensory information is
transmitted to the brain through the afferent vagus
nerve. Ghrelin, a peptide primarily produced in the stom-
ach, stimulates both feeding and growth hormone (GH)
secretion. How stomach-derived ghrelin exerts these
central actions is still unknown. Here we determined the
role of the gastric afferent vagal nerve in ghrelin’s func-
tions. Methods: Food intake and GH secretion were ex-
amined after an administration of ghrelin intravenously
(IV) to rats with vagotomy or perivagal application of
capsaicin, a specific afferent neurotoxin. We investi-
gated Fos expression in neuropeptide Y (NPY)-producing
and growth hormone–releasing hormone (GHRH)-pro-
ducing neurons by immunohistochemistry after admin-
istration IV of ghrelin to these rats. The presence of the
ghrelin receptor in vagal afferent neurons was assessed
by using reverse-transcription polymerase chain reaction
and in situ hybridization histochemistry. A binding study
on the vagus nerve by 125I-ghrelin was performed to
determine the transport of the ghrelin receptor from
vagus afferent neurons to the periphery. We recorded
the electric discharge of gastric vagal afferent induced
by ghrelin and compared it with that by cholecystokinin
(CCK), an anorectic gut peptide. Results: Blockade of the
gastric vagal afferent abolished ghrelin-induced feeding,
GH secretion, and activation of NPY-producing and
GHRH-producing neurons. Ghrelin receptors were syn-
thesized in vagal afferent neurons and transported to
the afferent terminals. Ghrelin suppressed firing of the
vagal afferent, whereas CCK stimulated it. Conclusions:
This study indicated that the gastric vagal afferent is the
major pathway conveying ghrelin’s signals for starvation
and GH secretion to the brain.
hrelin, a 28-amino acid peptide with an n-octanoyla-
G tion indispensable for its activity, was originally
discovered in human and rat stomach as a cognate en-
dogenous ligand for the growth hormone (GH) secreta-
gogue receptor.1 At present, ghrelin has been found in
fish, amphibians, reptiles, birds, and many mammals. It
is predominantly produced in a distinct type of endocrine
cell in the stomach.2 Ghrelin increases secretion of GH,
food intake, and body weight gain when administered
peripherally or centrally.1,3–10 Ghrelin activates agouti-
related protein-producing and neuropeptide Y (NPY)-
producing neurons localized in the arcuate nucleus of the
hypothalamus,6 –10 which is one of the brain regions of
primary importance in the regulation of feeding. Secre-
tion of ghrelin is up-regulated under conditions of neg-
ative energy balance such as starvation, insulin-induced
hypoglycemia, cachexia, and anorexia nervosa, whereas it
is down-regulated under conditions of positive energy
balance such as feeding, hyperglycemia, and obesity.11–15
GH secretion is also up-regulated under negative nutri-
tional status, whereas it is down-regulated under positive
nutritional status.16
The afferent fibers of the vagus nerve are the major
neuroanatomic linkage between the alimentary tract
and the nucleus of the solitary tract in the hindbrain,
where afferent input is integrated with descending
hypothalamic input and ascending output to the hy-
pothalamus is produced.17–20 Some meal-related me-
tabolites, monoamines, and peptides, as well as me-
chanical and chemical stimuli, transmit their satiety
signals to the nucleus of the solitary tract via the vagal
afferent or to the hypothalamus via the bloodstream.
Ghrelin had been thought to enter the brain across the
blood brain barrier. However, a recent study showed
that an intraperitoneal injection of ghrelin to totally
vagotomized mice did not stimulate food intake.8
Although ghrelin’s orexigenic activity may depend on
the vagus nerve, it has not been clarified how ghrelin
transmits information from the stomach to the brain.
Abbreviations used in this paper: GH, growth hormone; GHRH,
growth hormone-releasing hormone; ICV, intracerebroventricular; RT-
PCR, reverse-transcription polymerase chain reaction.
© 2002 by the American Gastroenterological Association
0016-5085/02/$35.00
doi:10.1053/gast.2002.35954
October 2002
Furthermore, it has not been known whether the vagus
nerve is involved in the regulation of GH secretion.
We studied food intake and plasma GH response
after an administration of ghrelin intravenously (IV)
to rats with gastric vagotomy or perivagal application
of a specific afferent neurotoxin, capsaicin.21 We also
examined the effects of these procedures on ghrelin-
induced c-fos expression, a marker of neuronal activa-
tion,22 in NPY and growth hormone–releasing hor-
mone (GHRH) neurons. We showed that ghrelin
suppressed the afferent discharge of the vagus nerve,
whereas cholecystokinin (CCK), an anorectic peptide
produced in the intestine,23 stimulated it. Finally, we
investigated the localization of the ghrelin receptor in
the vagal afferents. Stomach-derived ghrelin’s signals
for starvation and GH secretion are relayed to the
brain by means of the vagus nerve, which ultimately
interacts with the hypothalamus.
Materials and Methods
Animals
Male Wistar rats weighing 300 –350 g (Charles
River Japan, Inc., Shiga, Japan) were used in all experi-
ments. Rats were housed individually in plastic cages at a
constant room temperature in a 12-hour light (7 AM–7
PM)/dark cycle and were given standard laboratory chow and
water ad libitum. All procedures were performed in accor-
dance with the Japanese Physiological Society’s guidelines
for animal care. Anesthesia was performed by an intraperi-
toneal injection of sodium pentobarbital (Abbot Laborato-
ries, Chicago, IL). Rats were used as follows. First, bilateral
subdiaphragmatic vagotomy and gastric branch vagotomy
were performed as described previously.24 Second, the bi-
lateral subdiaphragmatic vagal nerves were exposed, then
loosely tied with a cotton string immersed with capsaicin
(Sigma Chemical Co., St. Louis, MO) dissolved in olive oil
(5% wt/vol).25 The cotton string was removed 30 minutes
later and the abdominal incision was closed. Third, intra-
cerebroventricular (ICV) cannulae were implanted into the
lateral cerebral ventricle.26 Proper placement of the cannu-
lae was verified at the end of the experiment by the admin-
istration of dye. Only animals that showed progressive
weight gain 4 days after cannulation were used in subse-
quent vagotomy, perivagal capsaicin application, or sham
operation.
Food Intake
Experiments were performed 1 week after vagotomy
or perivagal capsaicin application. Only rats that showed
progressive weight gain and food intake (body weight:
treated rats, 321.7 ⫾ 7.0 g; sham, 324.3 ⫾ 4.2 g; P ⬎ 0.7,
n ⫽ 10; dark phase food intake: treated rats, 23.4 ⫾ 1.0 g;
sham, 23.0 ⫾ 1.2 g; P ⬎ 0.7, n ⫽ 10) were used in feeding
ROLE OF VAGUS NERVE IN GHRELIN’S FUNCTIONS 1121
experiments. First, rat ghrelin (Peptide Institute, Inc.,
Osaka, Japan) was dissolved in 0.9% saline, and this solu-
tion (10 pmol–10 nmol/100 ␮L) was administered IV at 9
AM to naive rats fed ad libitum (n ⫽ 8 –10 per group).
Second, ghrelin (1.5 and 5 nmol/100 ␮L) was administered
IV to rats that had undergone bilateral subdiaphragmatic or
gastric branch vagotomy or capsaicin treatment, or to sham-
operated rats (n ⫽ 10 per group) after light anesthesia with
ether. Third, ghrelin (200 pmol/10 ␮L) was administered
ICV at 9 AM to rats that had undergone subdiaphragmatic
vagotomy or capsaicin treatment, or to sham-operated rats
(n ⫽ 10 per group). Fourth, rat des-acyl ghrelin (1.5 and 5
nmol/100 ␮L) was administered IV to naive rats (n ⫽ 10
per group). After injection, rats were immediately returned
to their cages, after which 2-hour food intake was measured.
We analyzed groups of data (mean ⫾ SEM) by using
analysis of variance (ANOVA) and post hoc Fisher test.
Growth Hormone Response
Rat ghrelin (1.5 nmol/100 ␮L) or rat GHRH (Sigma,
1.5 nmol/100 ␮L) was administered at 9 AM IV to rats that had
received perivagal capsaicin treatment, bilateral subdiaphrag-
matic vagotomy, or sham operation (n ⫽ 8 per group). Blood
samples (20 ␮L) were obtained from the tail vein at 0, 5, 10,
15, 20, 30, and 60 minutes after administration. The plasma
concentration of GH was determined with a Biotrak Rat GH
RIA kit (Amersham, Buckinghamshire, England).
Fos Expression
Ghrelin (1.5 nmol/100 ␮L) or saline was injected IV
to rats that had received bilateral subdiaphragmatic vagot-
omy, perivagal capsaicin treatment, or sham operation (n ⫽
3 per group) 90 minutes before transcardial perfusion with
fixative containing 4% paraformaldehyde. The brain was
sectioned into areas 40-␮m or 10-␮m thick. Immunohis-
tochemistry of Fos was performed as described.27 We sub-
jected some sections of the arcuate nucleus to double stain-
ing with anti-NPY antiserum (DiaSorin, Stillwater, MN;
final dilution 1/2000) or anti-GHRH antiserum (Chemicon
International, Inc., Temecula, CA; final dilution 1/1000).
Reverse-Transcription Polymerase Chain
Reaction and In Situ Hybridization
Total RNA was extracted from the nodose ganglion
and hypothalamus of Wistar rats with Trizol (Life Technolo-
gies, Fredrick, MD). Reverse-transcription polymerase chain
reaction (RT-PCR) for the ghrelin receptor was performed as
described.2 Some portions of the PCR products were then
sequenced by the BigDye Terminator Cycle Sequencing Sys-
tem (Applied Biosystems, Foster City, CA).
Nodose ganglia were cut with a cryostat in slices 12-␮m
thick. In situ hybridization of the ghrelin receptor messenger
RNA (mRNA) was performed by using 2 pairs of 33P 3⬘
end-labeled antisense oligonucleotide probes.28 A 100-fold
molar excess of each unlabeled probe served as the controls.
After washing, hybridization sections were exposed to autora-
1122 DATE ET AL. GASTROENTEROLOGY Vol. 123, No. 4

Figure 1. Effect of vagotomy or capsaicin treatment on ghrelin-induced food intake. (A) Two-hour food intake (mean ⫾ SEM) of free-feeding rats
after single administration of ghrelin IV (0.01–10 nmol). *P ⬍ 0.0001 vs. control vehicle. (B) Food intake of rats with bilateral subdiaphragmatic
or gastric branch vagotomy after single administration of ghrelin IV (1.5 and 5 nmol). Control rats underwent sham operation. *P ⬍ 0.0001. (C)
Food intake of rats with perivagal capsaicin application after single administration of ghrelin IV (1.5 and 5 nmol). *P ⬍ 0.001 vs. control. (D) Food
intake of rats with subdiaphragmatic vagotomy after single ICV administration of ghrelin (200 pmol). *P ⬍ 0.0001. (E) Food intake of rats with
perivagal capsaicin application after single ICV administration of ghrelin (200 pmol). *P ⬍ 0.0001. 䊐, saline; ■, ghrelin.

diography film (Hyperfilm; Amersham) for 21 days. The re-
sulting images were visualized by using an MCID imaging
analyzer (Imaging Research, Ontario, Canada). The sections
were coated with Kodak NTB3 emulsion (Eastman Kodak,
Rochester, NY) for autoradiography and further exposed for 60
days. After being developed in Kodak D-19 and fixed with
Fujifix (Fuji Film, Tokyo, Japan), they were counterstained
with H&E.
Vagal Ligation and Autoradiography
37°C with binding buffer (20 mmol/L HEPES, 150 mmol/L
NaCl, 5 mmol/L MgCl2 , 1 mmol/L ethylene glycol-bis(␤-
aminoethyl ether)-N,N,N⬘,N⬘-tetraacetic acid, and 0.1%
bovine serum albumin) for 60 minutes, after which 3
nmol/L [125I-Tyr29]-rat ghrelin was put onto the nerve and
incubated for 30 minutes. Nonspecific binding was deter-
mined in the presence of excess (3 ␮mol/L) unlabeled
ghrelin. The sections were exposed to an IP plate (Fuji Film)
for 24 hours, and analyzed in BAS-2000 (Fuji Film).

A crushing ligation of the vagus nerve of rats (n ⫽ Electrophysiologic Study

5) with suture thread was made 20-mm caudal to the
nodose ganglion. The vagus nerve was excised 16 hours
later, embedded in Tissue-Tek O.C.T. compound (Sakura
Finetechnical Co., Ltd., Tokyo, Japan), and frozen. Serial
sections (10 ␮m) were cut by using a cryostat along the
longitudinal axis of the nerve and they were mounted onto
gelatin-coated glass slides. The sections were incubated at
An isolated gastric vagal nerve filament of rats was
placed on a pair of silver wire electrodes to record afferent
discharges.29 Ghrelin (1.5 nmol), des-acyl ghrelin (1.5 nmol),
or CCK (Peptide Institute, Inc., 1.5 nmol) was administered
IV in the morning through the catheter inserted into the
inferior vena cava (n ⫽ 8 per group). Multiunit afferent nerve
October 2002 ROLE OF VAGUS NERVE IN GHRELIN’S FUNCTIONS 1123

Figure 2. Effect of capsaicin treatment or vagotomy on ghrelin-induced GH secretion. (A, B) Time courses of plasma GH concentration (mean ⫾
SEM) after administration of ghrelin IV to rats with (A) perivagal capsaicin application and (B) bilateral subdiaphragmatic vagotomy. (C, D) Time
courses of plasma GH concentration after administration of GHRH IV to rats with (C) perivagal capsaicin application and (D) gastric branch
vagotomy. *P ⬍ 0.01; **P ⬍ 0.001 vs. control. A and C: F, control; E, capsaicin; B and D: F, control; E, vagotomy.

discharge was recorded for 90 minutes after administration and
analyzed.29
Results
Peripheral Ghrelin Increases Food Intake
and Growth Hormone Secretion Via
the Afferent Vagal Nerve
We first tested various doses of ghrelin ranging
from 10 pmol to 10 nmol in a food intake experiment
(Figure 1A ). The lowest effective dose of ghrelin
administered IV was 1.5 nmol, which was used as a
standard dose in a subsequent series of experiments.
To investigate whether peripherally administered
ghrelin regulates feeding behavior via the abdominal
vagal nerve afferent, we evaluated the effect of vagot-
omy on ghrelin-induced food intake. Testing began 1
week after surgery when vagotomized rats had a stable
intake of food and water. A single administration of
ghrelin IV significantly increased food intake of con-
trol rats, but did not increase food intake in rats that
had undergone subdiaphragmatic or gastric branch
vagotomy (Figure 1B). There were no differences in
the effect of ghrelin given IV on feeding between the
subdiaphragmatic vagotomized rats and gastric branch
vagotomized rats. Therefore, we chose subdiaphrag-
matic vagotomy in the following experiments using
vagotomy. Next, because it is not clear if the critical
lesion involves gastric efferent fibers, afferent fibers, or
both, we applied a specific afferent neurotoxin, capsa-
icin, to the abdominal vagal trunk. Capsaicin abol-
ished feeding induced by administration of ghrelin IV
(Figure 1C ), suggesting that blockade of vagal affer-
1124 DATE ET AL. GASTROENTEROLOGY Vol. 123, No. 4

Figure 3. Localization of Fos expression in response to administration of ghrelin IV to rats that received capsaicin treatment or sham operation.
(A) Fos is found in neurons of the arcuate nucleus of sham-operated rats. Costaining of (B) Fos (blue-black) and NPY neurons (brown) and (C)
GHRH neurons (brown) in the arcuate nucleus of sham-operated rats. (D) No Fos expression is seen in response to ghrelin in capsaicin-treated
rats. 3V, third ventricle. Scale bars: A, D, 100 ␮m; B, C, 50 ␮m.

ents is the critical lesion. Because vagotomy and cap- received these treatments. An ICV administration of
saicin application may nonspecifically suppress feed- ghrelin equally increased food intake in vagotomized,
ing in response to ghrelin, we tested the orexigenic capsaicin-treated, and control groups (Figure 1D
effect of centrally administered ghrelin in rats that had and E ).
October 2002 ROLE OF VAGUS NERVE IN GHRELIN’S FUNCTIONS 1125

Next, we studied whether the GH-releasing activity of
peripheral ghrelin was also mediated by vagal sensory
function. The GH response to ghrelin was profoundly
attenuated by both capsaicin treatment (area under the
curve from 0 – 60 min, 5999.8 ⫾ 1264.9 ng/mL ⫻ hr vs.
1947.4 ⫾ 329.6 ng/mL ⫻ hr, P ⬍ 0.05) and gastric
branch vagotomy (area under the curve from 0 – 60 min,
8161.3 ⫾ 2014.0 ng/mL ⫻ hr vs. 3288.7 ⫾ 619.5
ng/mL ⫻ hr, P ⬍ 0.05) (Figure 2A and B), but the GH
response to GHRH was not affected by these 2 proce-
dures (Figure 2C and D).
Peripheral Ghrelin Activates Neuropeptide
Y– and Growth Hormone Releasing
Hormone–Containing Neurons Via
the Afferent Vagal Nerve
To establish the neuronal populations activated
by peripheral ghrelin, we investigated c-fos expression
after administration of ghrelin IV. Fos-immunoreactive
neurons were observed only in the arcuate nucleus of
control rats (Figure 3A). Of the 3 rats examined by
double immunohistochemistry, ghrelin induced Fos ex-
pression in 43% ⫾ 5% of NPY-containing neurons and
15% ⫾ 5% of GHRH-containing neurons (Figure 3B
and C). These results are consistent with a previous finding
that 51% of NPY neurons and 23% of GHRH neurons
expressed Fos after peripheral administration of GHRP-6,30
a synthetic GH-releasing hexapeptide that binds to the
ghrelin receptor.31 In both capsaicin-treated rats (Figure
3D) and vagotomized rats (data not shown), ghrelin did
not induce Fos in any neurons of the arcuate nucleus.
Ghrelin Receptor Is Present in the Vagal
Nodose Ganglion and Is Transported
to Afferent Terminals
We investigated the expression of the ghrelin
receptor in the vagal nodose ganglion by using RT-PCR,
direct sequencing, and in situ hybridization histochem-
istry. A ghrelin-receptor transcript product was found in
an mRNA sample isolated from the vagal nodose gan-
glion (Figure 4A). The DNA sequence of this PCR
product was consistent with that of the ghrelin receptor
(data not shown). Signals specific for 33P-labeled anti-
sense probes for the ghrelin receptor were found in the
area containing afferent neurons of the nodose ganglion
(Figure 4B and C). The signals disappeared with the
addition of an excess of unlabeled probes (Figure 4D).
Approximately 40% of neuronal cell bodies in the vagal
nodose ganglion had positive signals for the ghrelin
receptor probes (Figure 4E).
To study the transport of the ghrelin receptor in the
vagus nerve, we examined the effect of vagal ligation on
the accumulation of binding sites detected by using
125I-ghrelin. Binding sites of 125I-ghrelin accumulated in
the segments proximal to the ligature (Figure 4F and
G). This binding was abolished by unlabeled ghrelin
(Figure 4G).
Ghrelin Decreases Gastric Vagal
Afferent Activity
Ghrelin administered IV (1.5 nmol) significantly
decreased gastric vagal afferent activity (Figure 5A).
Administration of des-acyl ghrelin IV, which lacks the
n-octanoylation at Ser 3 that is essential for ghrelin’s
binding activity to the receptor,1 did not induce food
intake (2-hour food intake: 5 nmol des-acyl ghrelin,
0.28 ⫾ 0.13 g; vehicle, 0.22 ⫾ 0.08 g; P ⬎ 0.5) or GH
secretion (data not shown). Administration of des-acyl
ghrelin IV did not affect the afferent activity (Figure 5B).
These findings suggest the electrophysiologic specificity
of ghrelin on the gastric vagal afferent. In contrast,
administration of CCK IV significantly increased gastric
vagal afferent activity (Figure 5C).
Discussion
The vagus nerve is a cranial nerve that contains
both efferent and afferent fibers. Approximately 90% of
the vagus nerve fibers in the subdiaphragm are afferent
and are composed of unmyelinated, thin, capsaicin-sen-
sitive fibers.32 There are some afferent endings within the
gastrointestinal mucosa and submucosa that are more
optimally positioned to monitor luminal composition
and bioactive substances released from enteroendocrine

Š
Figure 4. Expression of ghrelin receptor mRNA in vagal nodose ganglion and binding of 125I-ghrelin in the vagus nerve. (A) Representative
electrophoretic analysis patterns of RT-PCR products of ghrelin receptor mRNA in the nodose ganglion and hypothalamus of rats. (B)
Photomicrograph of the nodose ganglion stained with H&E to visualize its cytoarchitecture. (C, D, and E) In situ hybridization for ghrelin receptor
mRNA. (C) Hybridization signals on the nodose ganglion (red). (D) No hybridization signals are seen on the nodose ganglion after addition of an
excess of unlabeled probes. (E) Bright-field photomicrograph of liquid emulsion autoradiography. Positive signals are present on cell bodies.
Scale bars: B, C, and D, 500 ␮m; E, 100 ␮m. (F ) Representative autoradiograph showing binding sites of 125I-ghrelin in the vagus nerve. (G)
Accumulation of 125I-ghrelin binding sites around the ligation of the vagus nerve. Binding of 125I-ghrelin is abolished by addition of an excess of
ghrelin. Values along the abscissa indicate distance (mm) from the ligature. Negative values correspond to areas proximal to the ligature and
positive values correspond to areas distal to the ligature. Data are expressed as mean ⫾ SEM (n ⫽ 5). *P ⬍ 0.02; **P ⬍ 0.005 vs. distal 2
mm. (G) F, 125I-ghrelin; E, cold excess.
1126 DATE ET AL.
Figure 5. Effects of administration of ghrelin IV, des-acyl ghrelin, and CCK on gastric vagal afferent discharge. (A) Ghrelin, (B) but not des-acyl
ghrelin, suppresses gastric vagal afferent activity. *P ⬍ 0.001; **P ⬍ 0.0001 vs. value at 0 minutes. (C) Stimulative effect of CCK on the gastric
vagal afferent activity. *P ⬍ 0.05; **P ⬍ 0.005 vs. value at 0 minutes. Representative data of gastric vagal afferent discharge rates are shown
in the upper figures. Vertical bar: 100 impulses/5 sec; horizontal bar: 30 minutes.
cells.33 Neural and humoral signals produced in the
gastrointestinal tract transmit messages for satiety and
starvation to the brain via the afferent vagal nerve, blood
circulation, or both. A large number of peptides that
regulate food intake come from the gut-brain group of
neuroenteric peptides. The existence of an orexigenic
peptide-based system in the periphery has yet to be
shown. Ghrelin is the first neuroenteric peptide that acts
as a starvation-signaling molecule in the periphery and
stimulates feeding after peripheral administration.
The present study showed that circulating ghrelin acts
predominantly on the stimulation of feeding and secre-
tion of GH via the gastric vagal afferents. Selective
chemical and surgical deafferentation of the gastric vagal
nerve blocked the food intake induced by peripheral
administration of ghrelin, but did not affect the food
intake induced by centrally administered ghrelin. Ghre-
lin administered IV induced Fos expression only in the
arcuate nucleus as reported previously,34 whereas cen-
trally administered ghrelin induced Fos expression in
various brain regions including several hypothalamic
nuclei, the dentate gyrus, the hippocampus, and the
cerebral cortex, as well as in the arcuate nucleus.7 These
findings imply that a neuronal pathway mediated by
peripheral ghrelin differs from a pathway mediated by
central ghrelin. Perivagal capsaicin treatment and vagot-
omy profoundly reduced GH secretion and canceled Fos
expression in GHRH neurons in rats given Ghrelin IV.
This treatment did not affect GHRH-induced GH se-
cretion. Ghrelin elicits GH secretion by a dual action, a
direct effect on the pituitary and modulation of GHRH
and somatostatin in the hypothalamus.1,35,36 Residual
GH secretion induced by ghrelin when vagal afferents are
GASTROENTEROLOGY Vol. 123, No. 4
blocked may be attributed to a direct pituitary response
via the systemic circulation.
Receptors in the vagus are synthesized at the cell
bodies and transported to the nerve terminals through
axonal transport. A prominent swelling of the vagus
immediately before its entrance into the cranial cavity is
known as the nodose ganglion, which is composed of
about 6000 neurons in the rat.37 Here we showed that
the ghrelin receptor was synthesized in vagal afferent cell
bodies and transported to the periphery. This may sug-
gest that there is a close proximity between ghrelin-
producing cells and vagal afferent terminals in the stom-
ach. Vagal afferent neurons have been known to produce
several bioactive peptides including substance P and
calcitonin gene–related peptide relative to feeding.38
Substance P and calcitonin gene–related peptide, which
are present in a large number of vagal afferent neurons,
suppress food intake in 24-hour fasted rats when admin-
istered ICV.39,40 Although whether or not substance P–
and calcitonin gene–related peptide– containing neurons
are sensitive to ghrelin has not been clarified, ghrelin’s
stimulative action on feeding may be exhibited partly
through the regulation of the substance P and calcitonin
gene–related peptide systems.
Vagal afferent fibers have a continuous low-frequency
spontaneous discharge that is modulated by sensory in-
puts.41,42 The present electrophysiologic studies verified
that the effective dose of ghrelin to stimulate feeding and
GH secretion suppressed gastric vagal afferent discharge.
In contrast, CCK, a gut-brain peptide that transmits a
satiety signal to the nucleus of the solitary tract via vagal
afferents,24,43,44 excited it as reported.23,45 Ghrelin may
October 2002
play a paracrine or endocrine role in the transduction of
afferent signals.
In addition to satiety information, the stomach ap-
prises the brain of ghrelin-derived hunger information
via vagal afferent. Ghrelin also provides a new insight
into the vagus-mediated GH secretion system. Ghrelin
and GH serve as anabolic signaling molecules during
energy depletion. Ghrelin production in the stomach
seems logical considering these anabolic effects. The
study of ghrelin will extend our understanding of how
energy balance and growth are controlled by the stomach.
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Received May 6, 2002. Accepted June 13, 2002.
Address requests for reprints to: Masamitsu Nakazato, M.D.,
Ph.D., Third Department of Internal Medicine, Miyazaki Medical Col-
lege, Kiyotake, Miyazaki, 889-1692, Japan. e-mail: nakazato@post.
miyazaki-med.ac.jp; fax: (81) 985-85-7902.
Supported in part by grants in aid from the Ministry of Education,
Culture, Sports, Science, and Technology, Japan, and the Ministry of
Health, Labor, and Welfare, Japan (to M.N.).
The authors thank M. S. Mondal, T. Hanada, K. Fukunaga, and R.
Matsuura for technical assistance.