European Journal of Pharmacology 561 (2007) 182 – 188

www.elsevier.com/locate/ejphar

Enhancement effects of nicotine on neurogenic contractile

responses in rabbit gastric fundus
Sevil Özger İlhan a , İsmail Mert Vural b,⁎, Ergin Dileköz c , Gökçe Sevim Öztürk b , Yusuf Sarioglu b
a
Refik Saydam Hygiene Center Presidency, School of Public Health, Ankara, Turkey
b
Department of Pharmacology, Medical School, Gazi University, 06510 Beşevler, Ankara, Turkey
c
Massachusetts General Hospital, Harvard Medical School Radiology Department Charlestown, MA 02129, USA
Received 27 July 2006; received in revised form 15 December 2006; accepted 21 December 2006
Available online 20 January 2007

Abstract

Nicotine, a nicotinic acetylcholine receptor agonist, plays a role in the modulation of neurotransmitter release following nerve stimulation in
both the central and the peripheral nervous system. Nitric oxide and prostaglandins modulate the release of various neurotransmitters in different
tissues. We aimed to investigate the effects of nicotine on neurogenic contractile responses via nicotinic acetylcholine receptors and, if a change
occurred, to investigate the effects of NW-nitro-L-arginine methyl ester (L-NAME) and indomethacin on this change in rabbit gastric fundus.
Electrical field stimulation (EFS)-evoked contractile responses were recorded from gastric fundus strips obtained from rabbits with an isometric
force displacement transducer. Nicotine was applied to preparations at varying concentrations. Then, the effects of hexamethonium, cadmium
(Cd2+), indomethacin, and L-NAME were tested on the EFS-evoked contractions in the presence of nicotine. Nicotine-induced transient
neurogenic contractions in a dose-dependent manner. Cd2+ and hexamethonium inhibited nicotine-induced transient neurogenic contractions, but
indomethacin and L-NAME produced no effect. In conclusion, nicotine increased EFS-evoked contractile responses, possibly by facilitating
neurotransmitter release from nerve terminals by a mechanism dependent on the influx of Ca2+ from voltage-gated Ca2+ channels via activation
of nicotinic acetylcholine receptors in isolated rabbit gastric fundus. Endogenous nitric oxide and prostaglandins do not play a physiological role
in the regulation of this neurotransmitter release.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Nicotine; Rabbit gastric fundus; Nicotinic acetylcholine receptor; Electrical field stimulation; Nitric oxide; Prostaglandins

1. Introduction (Todorov et al., 1991; Nedergaard and Schrold, 1977; Yokotani

et al., 2001).
Nicotine, an alkaloid isolated from the leaves of the tobacco It was previously demonstrated that, in peripheral tissues,
plant, acts as an agonist of nicotinic acetylcholine receptors, nicotine increases the release of various transmitters via
which are located in both the central and the peripheral nervous peripheral nicotinic acetylcholine receptors (Cuprian et al.,
system (Newman et al., 2002; McGehee et al., 1995; Todorov 2005; Yokotani et al., 2002, 2000; Schneider and Galligan,
et al., 1991). Nicotinic acetylcholine receptors are members of a 2000; Todorov et al., 1991; Rand and Li, 1992; Briggs and
superfamily of pentameric ligand-gated ion channels, and have Cooper, 1982). In guinea pig vas deferens, nicotine also
seventeen identified subunits. Nicotine is known to modulate increased neurotransmitter release induced by nerve stimulation
neurotransmitter release via nicotinic acetylcholine receptors, but (Todorov et al., 1991) Nicotine also induces transient neuro-
the underlying mechanisms of this action are poorly understood genic contractions in mouse vas deferens (Brain et al., 2001). In
another study, nicotine was shown to induce norepinephrine
release in rat stomach, due to activation of ganglionic nicotinic
⁎ Corresponding author. Gazi University, Faculty of Medicine, Department of
acetylcholine receptors localized at the celiac ganglia. This
Pharmacology, Beşevler 06510, Ankara, Turkey. Tel.: +90 312 202 6951, +90 effect of nicotine was sensitive to hexamethonium and was
312 202 46 22; fax: +90 312 212 46 47. inhibited by tetrodotoxin (Yokotani et al., 2000). Similarly, it
E-mail address: [email protected] (İ.M. Vural). was shown that, in both ganglionic vas deferens and guinea pig
0014-2999/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.ejphar.2006.12.033
 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188 183

ileum myenteric plexus, nicotine can modulate acetylcholine 2.3. Organ chamber experiments
release via nicotinic acetylcholine receptors (Cuprian et al.,
2005; Briggs and Cooper, 1982). Schneider and Galligan Each strip was mounted under 1 g isometric resting tension in
reported that presynaptic nicotinic acetylcholine receptors in an organ bath containing 15 mL Krebs−Henseleit solution
guinea pig ileum mediate the release of substance P and/or (composition in mmol/L: NaCl 118.0, KCl 4.7, CaCl2·2H2O
neurokinin A from myenteric neurons (Schneider and Galligan, 1.3, MgCl2·6H2O 0.5, Na2HPO4·2H2O 0.9, NaHCO3 24.9,
2000). Activation of nicotinic acetylcholine receptors by nico- glucose monohydrate 11.0). The pH of the solution was 7.4 after
tine mediates a Ca2+ influx through ligand-gated and/or voltage- bubbling with a mixture of 95% O2 and 5% CO2, and the solution
gated Ca2+ channels, which triggers the release of Ca2+ from was maintained at 37 °C. The tissues were allowed to equilibrate
intracellular calcium stores. This mechanism is thought to play for at least 1 h before experimental procedures. Isometric contrac-
an important role in nicotine-dependent effects on neurotrans- tions were evoked by EFS through a pair of platinum electrodes
mitter release (Brain et al., 2001; Dolezal et al., 1998; Shoop with an 8 Hz stimulation frequency by 10 s trains of impulses
et al., 2001; Sharma and Vijayaraghavan, 2001). delivered every 2 min. A stimulator (S48; Grass Instruments,
Nitric oxide modulates the release of various neurotransmit- Quincy, MA, USA) delivered 60 V pulses of 1 ms duration. EFS-
ters in different tissues, including acetylcholine release from evoked responses were recorded via Grass isometric force
hippocampal slices (Lonart et al., 1992) and mice ileum (Mang displacement transducers (Grass FT 03) connected to an ink
et al., 2002), γ-aminobutyric acid (GABA) and glycine release writing oscillograph (Grass 79 E) via a preamplifier. Thirty
from retina (Yu and Eldred, 2005), and norepinephrine release minutes after the EFS-evoked responses reached a steady state, to
from rat mesenteric arterial bed (Kolo et al., 2004). The test the contribution of the cholinergic component, the tissue was
underlying mechanisms by which nitric oxide modulates the treated with atropine (10− 6 M), a muscarinic receptor blocker,
release of these neurotransmitters remain unclear. Various theo- and neostigmine (10− 5 M), a reversible anticholinesterase drug.
ries have been suggested, targeting cyclic guanosine monopho- The effects of tetrodotoxin (3 × 10− 6 M), cadmium (Cd2+)
sphate (cGMP) (Guevara-Guzman et al., 1994), peroxynitrite (10− 4 M), hexamethonium (10− 5 M), NW-nitro-L-arginine methyl
(Ohkuma et al., 1995), Ca+2, and Na+ (Ohkuma et al., 1996) as ester (L-NAME) (10− 4 M), and indomethacin (10− 5 M) on the
mediators of the effects of nitric oxide on neurotransmitter EFS-evoked responses were also tested. To test the effects of
release. Prostaglandins are also known to modulate neurotrans- nicotine, different concentrations (10− 6 M, 3 × 10− 6 M, 10− 5 M,
mitter release. Prostaglandin E2 and arachidonic acid have been 3 × 10− 5 M, 10− 4 M) of nicotine were administered to the
shown to increase or decrease the release of neurotransmitters in preparations. To avoid any possible habituation effect or
various tissues, such as glutamate in rat hippocampus (Cunha tachyphylaxis, EFS was stopped after seven contractions, and
et al., 2004) and acetylcholine in the esophagus (Cheng et al., the preparations were washed for an hour in every 15 min.
2005) and frog neuromuscular junction (Arkhipova et al., 2006). Following washing, EFS was delivered again and the same
The aim of this study was to investigate the effects of experimental procedure was performed with the same tissue in the
nicotine on electrical field stimulation (EFS)-evoked neurogen- presence of hexamethonium (10− 5 M), Cd2+ (10− 4 M), L-NAME
ic contractile responses in rabbit gastric fundus, and to (10− 4 M), and indomethacin (10− 5 M). Antagonists/blockers
investigate the mechanism of any changes. The possible roles were added to the organ baths 30 min before the administration of
of nitric oxide and prostaglandins were also evaluated. nicotine.

2. Materials and methods 2.4. Drugs

2.1. Animals All drugs (nicotine, atropine sulfate, neostigmine, cadmium

chloride-1-hydrate, hexamethonium hydrochloride, tetrodotox-
Twenty New Zealand albino rabbits weighing 2.5−3.0 kg in, indomethacin, L-NAME hydrochloride were obtained from
were used for the experiments. All animals were kept under Sigma (St Louis, MO, USA). Stock solutions of drugs were
controlled temperature (23.2 °C) and humidity (55.5%) with a dissolved in distilled water, except for indomethacin, which was
14-hour light and 10-hour dark cycle. They were fed standard dissolved in 1% Na2CO3. Solutions were stored at −20 °C until
laboratory chow and given tap water. All experiments were use. The drugs were diluted in Krebs solution to the required
performed in accordance with the ethical regulations of the final concentration on the day of use. Vehicle controls showed
Helsinki Declaration. This study was approved by Gazi no effects.
University Ethics Committee for Animals.
2.5. Statistics
2.2. Tissues
Nicotine-induced increases were expressed as percentages of
Animals were sacrificed by exsanguination and their stom- the control and the average of seven EFS-evoked contractile
achs were rapidly excised, opened lengthwise, and emptied. responses. The value of the last contraction before the appli-
Adherent fat, gross connective tissues, and gastric mucosa were cation of nicotine was taken as the control value.
removed, and uniform longitudinal strips (15 mm × 3 mm) were Experimental values were expressed as the mean ± SEM.
prepared from the smooth muscle of the gastric fundus. Groups were compared statistically using general linear models
184 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188

of analysis of variance (ANOVA) followed by post hoc analysis

with the Bonferroni test.
P values of b 0.05 were considered to be statistically
significant.

3. Results

EFS evoked contractile responses in rabbit stomach. The

mean amplitude of the EFS-evoked contractile responses was
2.85 ± 0.42 g at a stimulation frequency of 8 Hz.
Tetrodotoxin (3 × 10− 6 M), a blocker of Na+ channels,
abolished EFS-evoked contractile responses in rabbit gastric
fundus strips (Fig. 1A).

3.1. Effects of drugs on neurogenic contractions of rabbit

gastric fundus strips

EFS-evoked contractile responses in rabbit gastric fundus

strips were abolished by atropine, a muscarinic receptor blocker,
at a concentration of 10− 6 M (Fig. 1B).
Neostigmine, a reversible anticholinesterase drug, increased
the amplitude of the EFS-evoked contractile responses in a

Fig. 2. Effects of different concentrations of nicotine (nic.) (10− 6 (n = 10);

3 × 10− 6 (n = 10); 10− 5 (n = 10); 3 × 10− 5 (n = 10); 10− 4 (n = 10)) on the mean of
seven EFS-evoked responses. Each point is expressed as a percentage of the
control and is given as the mean ± SEM (⁎, P b 0.05). (A) Typical traces showing
the effects of different concentrations of nicotine (nic.) (10− 5 M; 3 × 10− 5 M;
10− 4 M] on EFS-evoked responses (B).

dose-dependent manner (10− 6 M, 11.1%; 3 × 10− 6 M, 100%;

10− 5 M, 722.2%; P b 0.05) (Fig. 1C).

3.2. Effects of nicotine on neurogenic contractions of rabbit

gastric fundus strips

Nicotine increased the EFS-induced contractions transiently

Fig. 1. Typical traces showing the effects of tetrodotoxin (3 × 10− 6 M) (A), in a dose-dependent manner (3× 10− 6 M, 28.2 ± 10.4%; 10− 5 M,
atropine (10− 6 M) (B), and neostigmine (10− 6 M) (C) on EFS-evoked contractile 62.2± 8.5%; 3 × 10− 5 M, 112.5± 16.3%; 10− 4 M, 153.7 ± 16.8%;
responses. P b 0.05), but had no effect at a concentration of 10− 6 M
 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188 185

(Fig. 2A, B). Nicotine-induced enhancements were reproducible

and were not significantly changed during the second period of
EFS after washing. No tachyphylaxis was observed.
Nicotine had no contractile effects on non-stimulated prep-
arations at the concentrations listed above (10− 6 M to 10− 4 M).

3.3. Effects of drugs on nicotine-induced EFS-evoked transient

neurogenic contractions

Hexamethonium (10− 5 M, 75.73 ± 15.17%; P b 0.05) inhib-

ited nicotine-induced EFS-evoked transient neurogenic con-
traction increases (Fig. 3A) but did not alter EFS-induced
contractions.
Cd2+ (10− 4 M, 82.69 ± 10.34%; P b 0.05) inhibited nicotine-
induced EFS-evoked transient neurogenic contraction increases
(Fig. 3B). In two of ten strips, Cd2+ (10− 4 M) inhibited EFS-
induced contractions. These strips were excluded from the
study. In the other strips, Cd2+ (10− 4 M) did not alter EFS-
induced contractions.

3.4. Effects of L-NAME and indomethacin on nicotine-induced

EFS-evoked transient neurogenic contractions

To investigate the effects of nitric oxide and prostaglandins Fig. 4. Effects of L-NAME (10− 4 M) on EFS-evoked contractions (B) and
on nicotine-induced EFS-evoked contractile response increases, 10− 4 M nicotine-induced (nic.) EFS-evoked contraction increases (D) (A)
(n = 8). Each point is expressed as a percentage of the control and the average
the responses were repeated in the presence of L-NAME or of seven EFS-evoked contractile responses. All points are given as the mean ±
indomethacin. SEM. Typical traces showing the effects of L-NAME (10− 4 M) on EFS-
Although both L -NAME (Fig. 4A-B) and nicotine evoked responses and nicotine-induced (nic.) EFS-evoked contractile
(Fig. 4A-C) led to an increase in the amplitude of the EFS- responses (B).
evoked contractile responses (10− 4 M, 303.41 ± 33.95%),

L-NAME had no effect on nicotine-induced transient neu-

rogenic contractions (Fig. 4). The effect of nicotine in the
presence of L-NAME was not significantly different than the
sum of the effects of L-NAME and nicotine separately. In the
presence of L-NAME, nicotine had no contractile effects on
non-stimulated preparations.
Indomethacin had no significant effect on EFS-evoked
contractile responses or nicotine-induced transient neurogenic
contractions (Fig. 5).

Fig. 5. Effects of indomethacin (indo.) (10− 5 M) on EFS-evoked contrac-

Fig. 3. Effects of different agents (hexametonium (hexa.) 10− 5 M (A) (n = 8); tions and 10− 4 M nicotine-induced (nic.) EFS-evoked contraction increases
Cd+ 2 10− 4 M (B) (n = 8)) on EFS-evoked contraction increases. Each point is (n = 8). Each point is expressed as a percentage of the control and the
expressed as a percentage of the control and the average of seven EFS-evoked average of seven EFS-evoked contractile responses. All points are given as
contractile responses. All points are given as the mean ± SEM (⁎, P b 0.05). the mean ± SEM.
186 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188

4. Discussion nicotine on neurotransmitter release (Brain et al., 2001; Dolezal

et al., 1998; Shoop et al., 2001; Sharma and Vijayaraghavan,
Pharmacological analysis of EFS-induced mechanical 2001). In this study, the effect of Cd2+, which blocks the
responses in isolated smooth muscle strips is a useful in vitro presynaptic voltage-gated Ca2+ channels involved in the EFS-
technique for clarifying autonomic innervation. In the present evoked responses, was tested to confirm the involvement of
study, tetrodotoxin, a blocker of Na+ channels, abolished EFS- these channels in neuro-effector transmission. Contractions
evoked responses, suggesting that EFS-evoked responses are elicited by EFS and nicotine-induced transient neurogenic
induced by nerve stimulation. contractions were reduced by Cd2+ at a concentration of
The gastric fundus is supplied with cholinergic nerves that 10− 4 M. These results suggest that voltage-gated Ca2+ channels
produce contractions via muscarinic receptors and these are required for excitatory neuro-effector transmission in the
cholinergic nerves play an important role in the regulation of rabbit gastric fundus and play a role in nicotine-induced tran-
gastrointestinal motility (Hammer, 1980, Heitkemper and sient neurogenic contractions.
Marotta, 1983, Leclere and Lefebvre, 2002a,b). It was previously In this study, L-NAME caused an increase in the amplitude of
reported that various neurotransmitters (e.g. vasoactive intestinal the EFS-evoked contractile responses but did not alter nicotine-
peptide (VIP), nitric oxide, ATP) can interfere with acetylcholine induced transient neurogenic contractions, indicating that
at the presynaptic and/or postsynaptic levels (Lefebvre et al., endogenous nitric oxide released from non-adrenergic, non-
1992; Baccari et al., 1994). In this study, we used atropine and cholinergic nerves does not play a physiological role in the
neostigmine to evaluate the involvement of cholinergic activation regulation of the neurotransmitter release, but interferes with
in EFS-evoked contractions. Atropine abolished and neostigmine acetylcholine by functional antagonism in the rabbit gastric
enhanced EFS-evoked contractile responses, indicating domi- fundus. Previously, it was shown that non-adrenergic, non-
nance of the cholinergic system in rabbit gastric fundus. cholinergic nerves release nitric oxide in rat gastric fundus and
Nicotine, a non-specific nicotinic acetylcholine receptor this endogenous nitric oxide interferes with cholinergic neuro-
agonist, increased EFS-evoked transient neurogenic contractions transmission either at the postsynaptic level via functional
in a dose-dependent manner in our experiments. The transient antagonism or at the presynaptic level via inhibition of acetyl-
neurogenic contractions were reversible and nicotine did not choline release (Lefebvre et al., 1992). In tissues such as
cause any tachyphylaxis. The transient neurogenic contractions hippocampal slices, mouse ileum, rat mesenteric arterial bed,
were characterized by a rapid onset and an initial, transitory peak. and retina, it has been demonstrated that nitric oxide modulates
These transient neurogenic contractions were reproducible and the release of various neurotransmitters by an unclear pharma-
did not change significantly over 1 h. Hexamethonium, a non- cological process (Lonart et al., 1992; Mang et al., 2002; Kolo
specific nicotinic acetylcholine receptor antagonist, prevented the et al., 2004; Yu and Eldred, 2005). Various theories regarding
potentiation caused by nicotine but had no effect on EFS-induced this have been suggested. It was shown that release of GABA is
responses in the absence of nicotine, showing that nicotinic evoked by nitric oxide via two mechanisms: a Ca2+-dependent
acetylcholine receptors are responsible for this effect of nicotine. release system and an Na+-dependent uptake system (Ohkuma
This finding indicates that nicotinic receptors are not involved in et al., 1996). According to other studies, cGMP (Guevara-
the normal physiological activation of the stomach. Although Guzman et al., 1994) or peroxynitrite (Ohkuma et al., 1995)
nicotine has been shown to have a facilitating effect on neuro- may mediate the effects of nitric oxide on neurotransmitter
transmitter release related to its effect on nicotinic acetylcholine release. It has also been shown that nitric oxide may alter the
receptors in both central and peripheral neurons, little is known synaptic protein interactions that regulate neurotransmitter
about the mechanism of this action (McGehee et al., 1995; Rose release (Meffert et al., 1996). In our study, the presence of L-
et al., 1999; Wang et al., 2000; Brain et al., 2001). In our previous NAME and nicotine affected the basal tonus of three of eight
studies, nicotine-induced transient neurogenic contractions in strips. We speculate that the activation of nicotinic acetylcholine
rabbit myometrium (unpublished) and bladder (Vural et al., receptors by nicotine might mediate the release of a contractile
2006). Furthermore, activation of nicotinic acetylcholine recep- substance, as previously shown in guinea pig ileum (Schneider
tors in guinea pig ileum myenteric plexus stimulated the release and Galligan, 2000). In the absence of nitric oxide, this
of neurotransmitters including acetylcholine (Galligan, 1999) substance might effect basal tonus. However, the effect of
and in mouse isolated vas deferens acetylcholine release from nicotine on basal tonus in the presence of L-NAME needs
cholinergic nerve terminals was increased by activation of nico- further investigation.
tinic acetylcholine receptors (Cuprian et al., 2005). In a different Indomethacin, a non-selective cyclooxygenase inhibitor, did
study, nicotine induced norepinephrine release from rat stomach not affect EFS-evoked contractile responses or nicotine-induced
via ganglionic nicotinic acetylcholine receptors localized at the transient neurogenic contractions. This suggests that endoge-
celiac ganglia, and hexamethonium and tetrodotoxin abolished nous prostaglandin release is not involved in the regulation of
this effect (Yokotani et al., 2000). neurotransmitter release in the rabbit gastric fundus. In other
It has been shown in various studies that influx of Ca2+ studies, however, it has been shown that prostaglandin E2 and
through ligand-gated and/or voltage-gated Ca2+ channels, via arachidonic acid increase neurotransmitter release in frog
activation of nicotinic acetylcholine receptors by nicotine, neuromuscular junction (Arkhipova et al., 2006) and decreases
triggers the release of Ca2+ from intracellular calcium stores. neurotransmitter release in rat hippocampus and esophagus
This Ca2+ activation plays an important role in the effect of (Cunha et al., 2004; Cheng et al., 2005). Although prostaglandins
 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188 187

are not normally released by nerves, their presence facilitated Heitkemper, M.M., Marotta, S.F., 1983. Development of neurotransmitter
contractions in guinea pig bladder. This contractile effect of enzyme activity in the rat gastrointestinal tract. Am. J. Physiol. 244,
G58–G64.
prostaglandin E2 in guinea pig bladder is possibly mediated by Kolo, L.L., Westfall, T.C., Macarthur, H., 2004. Modulation of neurotrans-
the mobilization of Ca2+ to nerve terminals (Creed and Callahan, mitter release by NO is altered in mesenteric arterial bed of spon-
1989). taneously hypertensive rats. Am. J. Physiol, Heart Circ. Physiol. 287,
In conclusion, nicotine increased EFS-evoked contractile H1842–H1847.
Leclere, P.G., Lefebvre, R.A., 2002a. Characterization of pre- and postsynaptic
responses in isolated rabbit gastric fundus, possibly by facil-
muscarinic receptors in circular muscle of pig gastric fundus. Br. J.
itating neurotransmitter release from nerve terminals by al- Pharmacol. 135, 1245–1254.
tering Ca 2+ influx through voltage-gated Ca 2+ channels via Leclere, P.G., Lefebvre, R.A., 2002b. Presynaptic modulation of cholinergic
activation of nicotinic acetylcholine receptors. The nicotinic neurotransmission in the human proximal stomach. Br. J. Pharmacol. 135,
acetylcholine receptor subtypes involved in nicotine-induced 135–142.
neurotransmitter release need further investigation. Endoge- Lefebvre, R.A., De Vriese, A., Smits, G.J., 1992. Influence of vasoactive
intestinal polypeptide and NG-nitro-L-arginine methylester on cholinergic
nous nitric oxide and prostaglandins do not play a physio- neurotransmission in the rat gastric fundus. Eur. J. Pharmacol. 221,
logical role in the regulation of nicotine-induced acetylcholine 235–242.
release in isolated rabbit gastric fundus. Further studies are Lonart, G., Wang, J., Johnson, K.M., 1992. Nitric oxide induces
required to identify the roles of, for example, substance P and neurotransmitter release from hippocampal slices. Eur. J. Pharmacol.
VIP, and to elucidate the role of acetylcholine in the regulation 220, 271–272.
Mang, C.F., Truempler, S., Erbelding, D., Kilbinger, H., 2002. Modulation by NO
of acetylcholine release. of acetylcholine release in the ileum of wild-type and NOS gene knockout
mice. Am. J. Physiol.: Gasterointest. Liver Physiol. 283, G1132–G1138.
Acknowledgment Meffert, M.K., Calakos, N.C., Scheller, R.H., Schulman, H., 1996. Nitric
oxide modulates synaptic vesicle docking fusion reactions. Neuron 16,
This work was supported by Gazi University Unit of 1229–1236.
McGehee, D.S., Heath, M.J., Gelber, S., Devay, P., Role, L.W., 1995. Nicotine
Scientific Research Projects (Project number: 11/2003-09). enhancement of fast excitatory synaptic transmission in CNS by presynaptic
receptors. Science 269, 1692–1696.
References Nedergaard, O.A., Schrold, J., 1977. The mechanism of action of nicotine on
vascular adrenergic neuroeffector transmission. Eur. J. Pharmacol. 42,
Arkhipova, O.V., Grishin, S.N., Sitdikova, G.F., Zefirov, A.L., 2006. 315–329.
The presynaptic effects of arachidonic Acid and prostaglandin E(2) Newman, M.B., Arendash, G.W., Shytle, R.D., Bickford, P.C., Tighe, T.,
at the frog neuromuscular junction. Neurosci. Behav. Physiol. 36, Sanberg, P.R., 2002. Nicotine's oxidative and antioxidant properties in CNS.
307–312. Life Sci. 71, 2807–2820.
Baccari, M.C., Calamai, F., Staderini, G., 1994. Modulation of cholinergic Ohkuma, S., Narihara, H., Katsura, M., Hasegawa, T., Kuriyama, K., 1995.
transmission by nitric oxide, VIP and ATP in the gastric muscle. Nitric oxide-induced [3H] GABA release from cerebral cortical neurons is
NeuroReport 5, 905–908. mediated by peroxynitrite. J. Neurochem. 65, 1109–1114.
Brain, K.L., Trout, S.J., Jackson, V.M., Dass, N., Cunnane, T.C., 2001. Nicotine Ohkuma, S., Katsura, M., Chen, D.Z., Narihara, H., Kuriyama, K., 1996.
induces calcium spikes in single nerve terminal varicosities: a role for Nitric oxide-evoked [3H] gamma-aminobutyric acid release is mediated
intracellular calcium stores. Neuroscience 106, 395–403. by two distinct release mechanisms. Brain Res. Mol. Brain Res. 36,
Briggs, C.A., Cooper, J.R., 1982. Cholinergic modulation of the release of [3H] 137–144.
acetylcholine from synaptosomes of the myenteric plexus. J. Neurochem. Rand, M.J., Li, C.G., 1992. Activation of noradrenergic and nitrergic
38, 501–508. mechanisms in the rat anococcygeus muscle by nicotine. Clin. Exp.
Cheng, L., Cao, W., Fiocchi, C., Behar, J., Biancani, P., Harnett, K.M., 2005. Pharmacol. Physiol. 19, 103–111.
Platelet-activating factor and prostaglandin E2 impair esophageal ACh Rose, J.E., Westman, E.C., Behm, F.M., Jonhson, M.P., Goldberg, J.S., 1999.
release in experimental esophagitis. Am. J. Physiol.: Gasterointest. Liver Blockade of smoking satisfaction using the peripheral nicotinic antagonist
Physiol. 289, G418–G428. trimethaphan. Pharmacol. Biochem. Behav. 62, 165–172.
Creed, K.E., Callahan, S.M., 1989. Prostaglandins and neurotransmission Schneider, D.A., Galligan, J.J., 2000. Presynaptic nicotinic acetylcholine
at the guinea pig and rabbit urinary bladder. Pflugers Arch. 413, receptors in the myenteric plexus of guinea pig intestine. Am. J. Physiol.:
299–302. Gasterointest. Liver Physiol. 279, G528–G535.
Cunha, R.A., Ribeiro, J.A., Malva, J.O., 2004. Presynaptic kainate receptors Sharma, G., Vijayaraghavan, S., 2001. Nicotinic cholinergic signaling in
modulating glutamatergic transmission in the rat hippocampus are inhibited hippocampal astrocytes involves calcium-induced calcium release from
by arachidonic acid. Neurochem. Int. 44, 371–379. intracellular stores. Proc. Natl. Acad. Sci. 98, 3631–3632.
Cuprian, A.M., Solanki, P., Jackson, M.V., Cunnane, T.C., 2005. Cholinergic Shoop, R.D., Chang, K.T., Ellisman, M.H., Berg, D.K., 2001. Synaptically
innervation of the mouse isolated vas deferens. Br. J. Pharmacol. 146, driven calcium transients via nicotinic receptors on somatic spines.
927–934. J. Neurosci. 21, 771–781.
Dolezal, V., Schobert, A., Hertting, G., 1998. Presynaptic nicotinic receptors Todorov, L., Windisch, K., Shersen, H., Lajtha, A., Papasova, M., Vizi, E.S.,
stimulate increases in intraterminal calcium of chick sympathetic neurons in 1991. Prejunctional nicotinic receptors involved in facilitation of stimula-
culture. J. Neurochem. 65, 1874–1879. tion-evoked noradrenaline release from the vas deferens of the guinea-pig.
Galligan, J.J., 1999. Nerve terminal nicotinic cholinergic receptors on excitatory Br. J. Pharmacol. 102, 186–190.
motoneurons in the myenteric plexus of guinea pig intestine. J. Pharmacol. Vural, I.M., Ozturk, G.S., Ercan, Z.S., Sarioglu, Y., 2006. Effects of the nicotine
Exp. Ther. 291, 92–98. on the neurogenic contractile responses via nicotinic acetylcholine receptors
Guevara-Guzman, R., Emson, P.C., Kendrick, K.M., 1994. Modulation of in in rabbit bladder tissue: reactive oxygen species do not plays a role in this
vivo striatal transmitter release by nitric oxide and cyclic GMP. mechanism. Pharmacology online 2, 54–62.
J. Neurochem. 62, 807–810. Wang, M., Okada, S., Murakami, Y., Yokotani, K., 2000. Nicotine-induced
Hammer, R., 1980. Muscarinic receptors in the stomach. Scand. J. Gastro- noradrenaline release from the isolated rat stomach by activation of L- and
enterol., Suppl. 66, 5–11. N-type calcium channels. Jpn. J. Pharmacol. 83, 102–106.
188 ˙
S. Özger Ilhan et al. / European Journal of Pharmacology 561 (2007) 182–188

Yokotani, K., Wang, M., Okada, S., Murakami, Y., Hirata, M., 2000. Yokotani, K., Okada, S., Nakamura, K., 2002. Characterization of functional
Characterization of nicotinic receptor-mediated noradrenaline release from nicotinic acetylcholine receptors involved in catecholamine release from the
the isolated rat stomach. Eur. J. Pharmacol. 402, 223–229. isolated rat adrenal gland. Eur. J. Pharmacol. 446, 83–87.
Yokotani, K., Okada, S., Murakami, Y., Nakamura, K., 2001. Nicotinic receptors Yu, D., Eldred, W.D., 2005. Nitric oxide stimulates gamma-aminobutyric acid
involved in gastric noradrenaline release evoked by electrical stimulation of release and inhibits glycine release in retina. J. Comp. Neurol. 483,
the splanchnic nerve in rats. Eur. J. Pharmacol. 423, 149–155. 278–291.