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Seizure: Taha Gholipour, Mehdi Ghasemi, Kiarash Riazi, Majid Ghaffarpour, Ahmad Reza Dehpour
Seizure: Taha Gholipour, Mehdi Ghasemi, Kiarash Riazi, Majid Ghaffarpour, Ahmad Reza Dehpour
Seizure
journal homepage: www.elsevier.com/locate/yseiz
Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, P.O. Box 13145-784, Tehran, Iran
Iranian Center of Neurological Research, Tehran University of Medical Sciences, Tehran, Iran
c
Interdisciplinary Neuroscience Research Program, Tehran University of Medical Sciences, Tehran, Iran
d
Department of Physiology and Biophysics, University of Calgary, Calgary, Alberta, Canada T2N 4N1
b
A R T I C L E I N F O
A B S T R A C T
Article history:
Received 6 May 2009
Received in revised form 28 October 2009
Accepted 29 October 2009
There is some evidence that epileptic seizures could be induced or increased by 5-hydroxytryptamine (5HT) attenuation, while augmentation of serotonin functions within the brain (e.g. by SSRIs) has been
reported to be anticonvulsant. This study was performed to determine the effect of selective 5-HT3
channel/receptor antagonist granisetron and agonist SR57227 hydrochloride on the pentylenetetrazole
(PTZ)-induced seizure threshold in mice. The possible interaction of this effect with nitrergic system was
also examined using the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME)
and the NO precursor L-arginine. SR57227 (10 mg/kg, i.p.) signicantly increased the seizure threshold
compared to control group, while high dose granisetron (10 mg/kg, i.p.) proved proconvulsant. Coadministration of sub-effective doses of the 5-HT3 agonist with L-NAME (5 and 60 mg/kg, i.p.,
respectively) exerted a signicant anticonvulsive effect, while sub-effective doses of granisetron (3 mg/
kg) was observed to have a proconvulsive action with the addition of L-arginine (75 mg/kg, i.p.). Our data
demonstrate that enhancement of 5-HT3 receptor function results in as anticonvulsant effect in the PTZinduced seizure model, and that selective antagonism at the 5-HT3 receptor yields proconvulsive effects.
Furthermore, the NO system may play a role in 5-HT3 receptor function.
2009 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.
Keywords:
5-HT3 receptor
Nitric oxide (NO)
Clonic seizure threshold
Pentylenetetrazole
Mice
1. Introduction
Among seven known classes of receptors for serotonin (5hydroxytryptamine: 5-HT), the 5-HT3 receptor is unique as being a
ligand-gated ion channel.1 It is structurally similar to other
neurotransmitter-dependent ion channels which also includes the
nicotinic acetylcholine channel/receptor.2,3 Unlike other 5-HT
receptors, 5-HT3 receptor causes an early depolarization effect in
postsynaptic membranes by directly conducting Na+, K+ and Ca2+
ions.1,4 As shown in animal brains, serotonergic brainstem
projections have excitatory effects on target neurons expressing
5-HT3 receptors.5 These targets are generally a subset of inhibitory
interneurons in cerebral neocortex6 and hippocampus,5,79 resulting in an overall inhibitory effect despite the excitatory nature of
1059-1311/$ see front matter 2009 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.seizure.2009.10.006
18
2.4. Experiments
First, animals received an intraperitoneal (i.p.) injection of
different doses of SR57227 (5 or 10 mg/kg), or granisetron (3 or
10 mg/kg) 30 min prior to seizure threshold measurement. In the
control group, mice received identical volumes of isotonic saline.
Next, we investigated the effects of NO on interaction between
serotonin and seizures. Sub-effective doses of SR57227 with LNAME (60 mg/kg; 45 min prior to seizure, i.e. 15 min prior to
SR57227), or sub-effective doses of granisetron with L-arginine
(75 mg/kg; again 45 min prior to seizure) were co-administered,
seeking for any probable additive effect.
2.5. Data analysis
Data are presented as mean standard error of the mean
(S.E.M.). Students t-test or One-way analysis of variance (ANOVA)
followed by the TukeyKramer multiple comparisons were used,
where appropriate, to analyze the data. The signicance level was
dened as p < 0.05.
3. Results
3.1. Effect of different doses of granisetron on seizure threshold
Granisetron (0 (saline), 3 or 10 mg/kg, i.p.) was injected 30 min
prior to the seizure determination. One-way ANOVA followed by
post hoc comparisons demonstrated that granisetron at 10 mg/kg
could lower the threshold of PTZ-induced clonic seizure (P < 0.01;
Fig. 1). However, a lower dose of granisetron (3 mg/kg, i.p.) failed to
show this proconvulsant activity, and was thus selected as a subeffective dose.
3.2. Effect of different doses of SR57227 on seizure threshold
The 5-HT3 receptor agonist SR57227 (0 (saline), 5 or
10 mg/kg, i.p.) was injected to groups of mice 30 min before
the seizure experiment. Analysis using one-way ANOVA
followed by post hoc comparisons showed the anticonvulsive
effect of this agent at 10 mg/kg (P < 0.001), but not at 5 mg/kg
(P > 0.05; Fig. 1).
2.2. Chemicals
Granisetron was used as 1 mg/ml injectable solution (manufactured and marketed by Roche, Switzerland) and was diluted,
when needed, by saline solution. SR57227 (1-(6-chloro-2-pyridinyl)-4-piperidinamine) hydrochloride was purchased from Tocris
(UK). Pentylenetetrazole (PTZ), L-NAME, and L-arginine were
purchased from Sigma (UK). Drugs were all dissolved in sterile
physiological saline. All injections were done in the volume of
10 ml/kg of the bodyweight of the mice. PTZ was prepared in
physiological saline as 1% solution.
2.3. Seizure threshold determination
The threshold of PTZ-induced seizures was measured by an
infusion of PTZ into the tail vein of freely moving mice at a constant
rate of 0.6 ml/min via a 30-gauge needle, connected by a
polyethylene tube to a Hamilton microsyringe.29 Minimal dose
of PTZ (mg/kg) required to induce general clonus was recorded. The
general clonus was characterized by forelimb clonus followed by
whole body clonus. As such, seizure threshold is dependent on PTZ
dose administered and time-related.3032
Fig. 2. Effects of different doses of L-arginine (50, 75, and 100 mg/kg, i.p.) on
pentylenetetrazole (PTZ)-induced seizure threshold in mice. L-Arginine was
administered intraperitoneally 45 min prior to seizure threshold determination.
Data represent mean S.E.M. of 68 mice. **P < 0.01 compared to control group
receiving normal saline (shown as dose 0).
19
Fig. 3. Additive effect of granisetron and L-arginine when co-administered in subeffective doses (3 and 75 mg/kg, i.p., respectively). Both drugs were administered
intraperitoneally; L-arginine 45 min and granisetron 30 min prior to seizure
threshold determination. In appropriate groups, normal saline was injected as
control. ***P < 0.001 compared to control group receiving no drug.
Fig. 4. Additive effect of SR57227 and L-NAME when co-administered in subeffective doses (5 and 60 mg/kg, i.p., respectively). Both drugs were administered
intraperitoneally; L-NAME 45 min and SR57227 30 min prior to seizure threshold
determination. In appropriate groups, normal saline was injected as control.
***P < 0.001 compared to control group receiving no drug.
20
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