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SEX DIFFERENCE IN SUSCEPTIBILITY TO PICROTOXIN-INDUCED

SEIZURES IN RATS FOLLOWING OCTREOTIDE
Meliha Tan a; Nuri Ihsan Kalyoncu a; Uner Tan a
a
BlackSea Technical University Medical School, Department of Neurology Trabzon, Turkey.

Online Publication Date: 01 August 2002

To cite this Article Tan, Meliha, Kalyoncu, Nuri Ihsan and Tan, Uner(2002)'SEX DIFFERENCE IN SUSCEPTIBILITY TO
PICROTOXIN-INDUCED SEIZURES IN RATS FOLLOWING OCTREOTIDE',International Journal of Neuroscience,112:8,903 — 911
To link to this Article: DOI: 10.1080/00207450290025914
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 Intern. J. Neuroscience, 112:903–911, 2002
Copyright  2002 Taylor & Francis
0020-7454/02 $12.00 + .00
DOI: 10.1080/00207450290025914

SEX DIFFERENCE IN SUSCEPTIBILITY

TO PICROTOXIN-INDUCED SEIZURES
IN RATS FOLLOWING OCTREOTIDE

MELIHA TAN
BlackSea Technical University
Medical School, Department of Neurology
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Trabzon, Turkey

NURI IHSAN KALYONCU

BlackSea Technical University
Medical School
Department of Pharmacology
Trabzon, Turkey

UNER TAN
BlackSea Technical University
Medical School
Department of Physiology
Trabzon, Turkey

It was previously reported that females with proestrous are more sus-
ceptible to picrotoxin-induced epileptic seizures than males, provided
that the estrous cycle is taken into consideration (Tan & Tan, 2001).
The sex difference in susceptibility to picrotoxin-induced seizures was
reconsidered in the present study using rats with a previous octreotide
(a stable somatostatin analogue) injection, as somatostatin is known to
be involved in epileptic phenomena. Contrary to rats without octreotide,
males were more susceptible to picrotoxin-induced seizures than females.
Statistical analysis indicated that only males showed decreased latencies

Address correspondence to Prof. Dr. Uner Tan, BlackSea Technical University, Medical
School, Department of Physiology, Trabzon, Turkey. E-mail: unertan@meds.ktu.edu.tr

903
 904 M. Tan et al.

to seizures and increased seizure frequencies following octreotide;

females were not affected by this experimental procedure. It was sug-
gested that as a GABA antagonist, somatostatin may contribute to the
occurrence of epileptic discharges in nervous system, creating a sex
difference to epileptic seizures. These results may have implications for
understanding the epileptic mechanisms, with possible therapeutic con-
sequences.

Keywords epilepsy, octreotide, picrotoxin, rat, sex difference, somatostatin

The scientific literature indicates that there is a sex difference in

susceptibility to epileptic seizures. However, where the difference
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lies has remained controversial and unresolved (see Tan & Tan,
2001). Using picrotoxin, an antagonist of the Cl– channel at the
GABAA receptor, Thomas (1990) reported a sex-associated suscep-
tibility to picrotoxin-induced myoclonic, focal, akinetic, and GTC
seizures in rats. At low doses, females had significantly shorter la-
tencies to myoclonic and GTC convulsions; at high picrotoxin doses,
the pattern of relative susceptibility of males and females to myo-
clonic seizures was reversed, with males having shorter latencies
than females. Thomas (1990) stated that significant sex differences
in seizure susceptibility exist, but only for specific seizure catego-
ries, and that the relative susceptibility of males and females de-
pends upon the dose of picrotoxin tested. On the other hand, there
is no significant difference in susceptibility to picrotoxin-induced
seizures between gonadectomized males and females (Schwartz-
Giblin et al., 1998): picrotoxin is subconvulsive for male rats but
convulsive for female rats (Pericic et al. (1985); female cats and
rats are more susceptible to seizures than male cats and rats (Pericic
et al., 1986).
Tan and Tan (2001) reconsidered the sex-related difference in
susceptibility to picrotoxin-induced epileptic seizures with relation
to the estrous cycle of the female rats. They found no significant
sex differences in the mean latencies to picrotoxin-induced seizures:
proestrous-females had the shortest latencies compared to males and
estrous-females. The authors concluded that there is a sex differ-
ence in susceptibility to seizures in rats, provided that the estrous
cycle is taken into account. Conversely, Dagci et al. (2002) studied
 Sex and Epileptic Seizures 905

wet dog shakes induced by kainic acid followed by epileptic sei-

zures induced by octreotide, a stable somatostatin analogue. The
authors discovered that octreotide increased the number of wet dog
shakes and decreased the number of convulsions, particularly in
males. Accordingly, we have hypothesized that the sex difference in
susceptibility to epileptic seizures would disappears under the so-
matostatin analogue octreotide. This hypothesis was tested in the
present work.

MATERIALS AND METHODS

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The experiments were performed on 10 male and 10 female Sprague-

Dawley rats, aged 70–90 days. The rats were housed in groups
of 3–4 and maintained on a 12-h light/dark cycle before testing;
only the cycling female rats (proestrous) were included in the ex-
periments. On the day of testing, rats were weighed and smears
were taken by a blunt plastic Eppendorf tip on the vaginal opening.
Pairs of males and females were tested at the same time in different
glass jars.
To begin, 1 mg/kg octreotide (Sandostatin) was intraperitoneally
(i.p.) injected into a pair of rats (one male and one female). Thirty
min later, picrotoxin dissolved in saline was administered i.p. (4
mg/kg), to induce epileptic seizures. Animals were then placed into
a plexiglas cage and observed for at least 60 min. Latencies to the
various seizure types were determined using a chronometer. Ani-
mals first entered a state of akinesis, characterized by motionless
posture and loss of righting ability (akinetic seizure). This was fol-
lowed by focal seizures with some tonic or clonic seizures in the
limbs, usually following an arching of the head and trunk. Myo-
clonic seizures were then started with quick, whole-body twitches
and jerks, followed by generalized tonic-clonic (OTC) epileptic
seizures. Some of the rats entered into status epilepticus following
the GTC seizures, which usually ended with exitus.
The statistical analysis was performed using the statistical pack-
age SPSS for Windows (V. 9.0). The effect size (e) was always
computed, if the differences were found to be statistically signifi-
cant.
 906 M. Tan et al.

RESULTS

Sex and Latencies of Seizures

Table 1 presents the mean latencies for various seizure types in the
male and female rats. ANOVA indicated that there was no sig-
nificant difference between the latencies of the akinetic seizures
(F1,18 = 1.06, p > .30). The mean latency of the focal seizures
was significantly longer in females than males (F1,18 = 5.59, p < .05,
e = .24), as was the myoclonic seizures (F1,14 = 11.20, p = .005, e =
.44); there was no significant difference between the mean laten-
cies of the GTCs (F1,16 = 2.48, p > .10) and status (F1,16 = 0.06, p >
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.80). The mean latency for exitus was found to be longer in females
than males, but the difference did not reach the traditional signifi-
cance level (F1,16 = 3.88, p = .06, e = .20). The mean latencies for
exitus were 24.0 ± 4.7 min and 29.1 ± 6.3 min for males and fe-
males, respectively.

Sex and Frequency of Seizures

Table 2 presents the mean frequencies for the akinetic, focal, myo-
clonic, and GTC seizure types in the male and female rats. There
was no significant sex difference in the frequencies of the akinetic
seizures (F = 0.15, p > .70). The frequency of the focal seizures
were significantly higher in females than males (F = 9.57, p < .01).
The mean frequency of the myoclonic seizure was found to be higher
in males than females, but the difference did not reach the tradi-
tional significance level of p = .05 (F = 3.82, p = .08). The mean

TABLE 1. Mean latencies for seizure types in male and female rats

Akinetic Focal Myoclonus GTC

Rats N Mean SD N Mean SD N Mean SD N Mean SD

MALE 10 2.85 1.42 10 5.23 0.64 10 5.87 1.96 10 8.06 1.14

FEMALE 10 3.84 2.69 10 7.82 3.41 6 15.08 8.52 8 10.11 3.94
MALEPIC 22 4.18 1.94 22 10.97 4.23 22 13.41 5.26 22 14.14 3.37
FEMALEP 22 3.17 1.10 22 9.13 5.69 22 12.27 6.86 22 15.02 9.26

Note. MALE and FEMALE: under octreotide + picrotoxin; MALEPIC and FEMALEP: only under picro-
toxin.
 Sex and Epileptic Seizures 907

TABLE 2. Mean frequencies of seizures in male and female rats

Akinetic Focal Myoclonic GTC

Rats Mean SD Mean SD Mean SD Mean SD

MALE 6.55 5.55 3.85 7.06 16.38 20.68 250.51 136.41

FEMALE 7.67 7.14 44.55 41.56 1.84 3.51 72.64 90.56

frequency for the GTC seizures was significantly higher in males

than females (F = 10.00, p < .01).

Comparison of Rats with Octreotide + Picrotoxin

with Those Having Only Picrotoxin
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Akinetic Seizures
The mean latencies to the various seizure activities are presented in
Table 1 (MALEFIC and FEMALEP). Statistical analysis indicated
that the mean latency for the akinetic seizure was significantly shorter
in the male rats with picrotoxin + octreotide (P + O) than the male
rats with picrotoxin only (t = 2.2, df = 30, p < .05). There was,
however, no significant group difference between the mean
latencies of the focal seizures for the female rats (t = 1.01, df = 30,
p > .30).

Focal Seizures
The mean latency was significantly shorter in the P + O males than
the P males (t = 4.23, df = 30, p < .001). The difference between the
mean latencies of the P + O females and P females was found to be
statistically nonsignificant (t = 0.67, df = 30, p > .50).

Myoclonic Seizures
The mean latency for the myoclonic seizures was significantly shorter
in the O + P males than the P males (t = 4.36, df = 30, p < .001).
There was, however, no significant difference between the mean
latencies of the myoclonic seizures for the P + O and P females (t =
1.00, df = 30, p > .30).
 908 M. Tan et al.

GTC Convulsions
The mean latency was found to be significantly shorter in P + O
males than P males (t = 5.52, df = 30, p < .001). The difference
between the mean latencies for the P + O females and P females
was, however, statistically nonsignificant (t = 1.60, df = 30, p >
.10).

Status Epilepticus
The mean latency for the P + O males was not significantly differ-
ent from that for the P males (t = 1.63, df = 30, p > .10); the mean
latency for the P + O females was also not significantly different
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from that for the P females (t = 0.78, df = 22, p > .40).

Exitus
The mean exitus latency was significantly shorter in the P + O
males than that for the P males (t = 8.55, df = 22, p < .001). There
was, however, no significant difference between the mean latencies
for the P + O females and the P females.

DISCUSSION

Tan and Tan (2001) previously reported that proestrous female rats
are more susceptible to picrotoxin-induced epileptic seizures than
male rats. Dagci et al. (2002) found, conversely, that octreotide, a
stable somatostatin analogue, suppressed kainic acid seizures, espe-
cially in the male rats. In the present work, we tested picrotoxin-
induced seizures in octreotide-treated rats, and discovered a reversal
of the picrotoxin effects concerning the sex difference. In particular,
male rats had usually shorter latencies to picrotoxin-seizures than
the female rats, and were more susceptible to epileptic seizures than
females, in contrast to the picrotoxin-induced seizures without octreo-
tide. Mean seizure frequencies were also higher in males than fe-
males, except the focal seizures, indicating different mechanisms of
octreotide actions in the rat brain. Generally speaking, it may be
 Sex and Epileptic Seizures 909

concluded that, under octreotide, males become more susceptible

to picrotoxin seizures than females, contrary to picrotoxin-induced
seizures without octreotide, as shown by Tan and Tan (2001).
Interestingly enough, the latencies for the picrotoxin-induced epi-
leptic seizures were significantly reduced by somatostatin, but only
in the male rats; the mean latencies were not significantly different
between the P + O females and the P females. As such, somatosta-
tin appears to affect cerebral circuits differentially in males and
females. Dagci et al. (2002) have also reported a similar sex-depen-
dent action of somatostatin on kainate-induced seizures in rats, where
somatostatin reduced the epileptic activity, especially in male rats.
Sex difference seems to be close to our results, but these investiga-
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tors have reported that somatostatin decreased the number of kainite-

induced convulsions, particularly in male rats (suppressive effect),
contrary to our results indicating that somatostatin increased the
number of convulsions and decreased the mean latencies of epilep-
tic seizures, but only in the male rats (activational effect).
Consistent with the report by Dagci et al. (2002), somatostatin is
generally found to be suppressive and protective for the epileptic
seizures. For instance, cortistatin similar to somatostatin exhibits an
anticonvulsive effect against kainate-induced seizures in rats (Braun
et al., 1998). Tallent and Siggins (1999) have studied the effects of
somatostatin on the hippocampal epileptiform activity in the rat hip-
pocampal slice preparation and found that somatostatin markedly
reduces the intensity of evoked epileptiform after discharge and the
frequency of spontaneous bursts in both CA1 and CA3. The authors
concluded that somatostatin receptors could be a unique target for
treatment of limbic seizures. However, this study does not provide
an answer to the question of whether this effect would be the same
in vivo. However, somatostatin is shown to have an inhibitory role
in hippocampal excitability and anticonvulsant properties in experi-
mental models of seizures (see Vezzani & Hoyer, 1999). The loss
of hilar somatostatinergic interneurons is the best documented
and most consistent interneuron deficit; somatostatin may be a
weak anticonvulsant in mice, but its loss alone may not account for
seizures in temporal lobe (Buckmaster et al., 2002). These studies
are consistent with the suppressive action of somatostatin on the
epileptic activity. However, there are no studies concerning the sex
 910 M. Tan et al.

difference in its actions. These results implicating an antiepileptic

role of somatostatin are inconsistent with our results, since we ob-
served a decrease in the latencies and an increase in the frequencies
of the picrotoxin-induced seizures in somaostatin treated rats.
On the other hand, there are also studies indicating a proconvul-
sive action of somatostatin, consistent with our results, but they also
did not consider the sex differences. It was reported that somato-
statin inhibits GABA-mediated synaptic transmission via a presyn-
aptic mechanism in cat and rat thalamocortical neurons, facilitating
the occurrence of seizures (Leresche et al., 2000); somatostatin has
excitatory effects on neurons in children with febrile seizures (Hirai
& Seki, 2000); it is elevated in cortex, striatum, and amygdala dur-
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ing electrically and pharmacologically kindled models, indicating

an active participation in the process of seizure susceptibility (see
Nagaki et al., 2000). There are more studies indicating the recruit-
ment of somatostatin-synthesizing neurons during seizures (e.g., Pretel
et al., 1996; Vezzani et al., 1996a, b).

CONCLUSIONS

It was previously reported that female rats in proestous are more

susceptible to picrotoxin-induced epileptic seizures than male rats
(Tan & Tan, 2001). Since somatostatin is involved in epileptic phe-
nomena, the sex difference in susceptibility to picrotoxin-induced
seizures was reconsidered in rats under octreotide, a stable soma-
tostatin analogue, treatment. Interestingly, the sex role reversed under
somatostatin: males were more susceptible to seizures than females
and females were not affected by this treatment. These results sug-
gest that as a GABA antagonist, somatostatin may trigger or en-
hance epileptic seizures, depending upon the sex. This would have
implications for understanding of the epileptic mechanisms with
possible therapeutic consequences.

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