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Changes of cortical epileptic afterdischarges

after status epilepticus in immature rats

ARTICLE in EPILEPSY RESEARCH · MARCH 2008

Impact Factor: 2.19 · DOI: 10.1016/j.eplepsyres.2007.11.008 · Source: PubMed

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Epilepsy Research (2008) 78, 178—185

journal homepage: www.elsevier.com/locate/epilepsyres

Changes of cortical epileptic afterdischarges after

status epilepticus in immature rats
Grygoriy Tsenov, Hana Kubová, Pavel Mareš ∗

Institute of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, CZ-14220 Prague 4, Czech Republic

Received 7 August 2007; received in revised form 20 November 2007; accepted 22 November 2007

KEYWORDS Summary Status epilepticus (SE) in developing rats leads to neuronal degeneration in many
brain structures including neocortex but the functional consequences of cortical damage were
Pilocarpine;
studied only exceptionally.
Status epilepticus;
Lithium—pilocarpine SE was elicited in 12- (P12) and 25-day-old (P25) rats, convulsions were
Cortical
interrupted after 2 h by paraldehyde. Cortical electrodes were implanted 3, 6, 9, 13 and/or
afterdischarges;
26 days after SE. Low-frequency stimulation of sensorimotor cortex was repeated with at
Ontogeny;
least 10-min intervals with a stepwise increasing intensity (0.2—14 mA). Thresholds for move-
Rat
ments elicited by stimulation, spike-and-wave afterdischarges (ADs), clonic seizures, mixed
ADs (transition into a limbic type of ADs) and recurrent ADs as well as duration of ADs were
evaluated.
The first three phenomena were not influenced by SE with the exception of lower thresholds
for movements during stimulation. Transition into limbic seizures and recurrent seizures were
delayed in both age groups and threshold intensities for limbic ADs were at some intervals
higher in SE than in control animals. Duration of ADs was changed only at short intervals after
SE; it was shortened at 3 and 6 days in P25 and 3 days in P12 rats, respectively. P12 group then
exhibited a transient increase in duration of ADs 6 days after SE.
Our results did not prove a higher cortical excitability after SE in either age group. On the
contrary, there were some signs of a decreased excitability.
© 2007 Elsevier B.V. All rights reserved.

Introduction epileptogenesis. It was demonstrated that it leads to degen-

Lithium—pilocarpine-induced status epilepticus (SE) in rats
represents a model of brain damage inducing chronic
∗ Corresponding author. Tel.: +420 24106 2495;
fax: +420 24106 2488.
E-mail address: maresp@biomed.cas.cz (P. Mareš).
eration of neurons in many brain structures in adult rodents
(Turski et al., 1983; Olney et al., 1986). It is clear that
lithium—pilocarpine SE induced at early stages of develop-
ment may also result in neuronal degeneration but these
changes are age-dependent (Sankar et al., 1998, 2000;
Kubová et al., 2001, 2002, 2004). Other models of SE
may lead to different patterns as well as age profile
of damage in the developing brain (kainic acid-induced
SE—–Sperber et al., 1991; Leite et al., 1996; electrical

0920-1211/$ — see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.eplepsyres.2007.11.008
Changes of cortical epileptic ADs after SE in rats
stimulation-induced SE—–Sankar et al., 2000), i.e. neuronal
degeneration is also model-dependent (Sankar et al., 1998,
2000). There are descriptions of damage in limbic struc-
tures after lithium—pilocarpine SE (Cavalheiro et al., 1987;
Sankar et al., 1997; Nairismagi et al., 2006), age-dependent
changes in subunit composition of GABA-A (decrease of ␣1
subunit after SE induced during adulthood in contrast to an
increase if SE was elicited at postnatal day 10 or 20—–Zhang
et al., 2004; Brooks-Kayal, 2005; Raol et al., 2006) and exci-
tatory amino acid receptors (Porter et al., 2006) in dentate
gyrus. Functional studies were also focused on hippocam-
pal formation. Among behavioral studies Morris water maze
(test of spatial memory dependent on hippocampus—–Morris,
1984) is most common (Mareš, 2007), studies of excitability
are also focused on limbic structures in a pilocarpine model
of SE (amygdala kindling—–Cilio et al., 2003) as well as in
other developmental models (Jensen et al., 1992, 1998; Lee
et al., 1995; Chen et al., 1999; Dube et al., 2000; Jensen
and Baram, 2000; Cilio et al., 2003; Zhang et al., 2004).
Recently morphological changes of thalamus were described
in detail (Kubová et al., 2001, Druga et al., 2005) but neo-
cortical damage was sometimes shortly mentioned (Turski
et al., 1983; Cavalheiro et al., 1987) but not studied in
detail in spite of the dominant role of cerebral cortex in
childhood epilepsies (Aicardi, 1998). Irreversibly damaged
neurons were demonstrated after lithium—pilocarpine SE in
cortex of 3- and 4-week-old rats (Sankar et al., 1997). Activ-
ity of cortical neurons analyzed in Cavalheiro’s (Sanabria et
al., 2002) and as well as in our laboratory (Dóczi et al., 2003)
revealed changes in the firing pattern of nerve cells after
SE induced at early developmental stages. Benzodiazepine
receptors in the cerebral cortex also exhibit age-dependent
changes: an increase 1 week after SE elicited at P12 and a
decrease 3 months after SE induced in adult rats (Rocha et
al., 2007). In connection with these findings we started a
detailed study of morphological and functional changes of
sensorimotor cortical area. Our first study described oppo-
site changes of transcallosal (interhemispheric) responses
in the sensorimotor cortex of rats exposed to SE at the age
of 12 and/or 25 days (Mareš et al., 2005). There were no
significant differences 3, 6 and 9 days after SE in compar-
ison with control rats. Changes were observed only 2 and
more weeks after SE, amplitude of responses was lower in
animals undergoing SE at P12 than in corresponding con-
trols whereas input—output curve was steeper in rats with
SE at P25 than in their control group. Cortical excitability
was also studied by means of rhythmic stimulation: series of
five stimuli can induce frequency potentiation and/or fre-
quency depression. Our study of these phenomena did not
reveal any marked changes of cortical excitability after SE
elicited at the age of 12 and/or 25 days (Tsenov and Mareš,
2007).
Cortical epileptic afterdischarges represent another pos-
sibility how to study cortical excitability (Mareš and Kubová,
2006). This model elicited by low-frequency stimulation
of sensorimotor cortical area is frequently used in our
laboratory in adult as well as immature rats (Lojková
et al., 2006; Mareš and Šlamberová, 2006). It allows to
study four different phenomena, three of them epileptic
in nature: movements directly elicited by stimulation, EEG
afterdischarges of the spike-and-wave type, clonic seizures
accompanying this type of afterdischarges, and with high
179
stimulation intensities a transition from spike-and-wave into
another type of afterdischarges identical with that induced
by hippocampal stimulation. Ontogeny of these phenomena
was described in detail (Mareš et al., 2002) so that longitu-
dinal study after SE could demonstrate not only a marked
pathology but also a modification of brain development.
Methods
Animals
Male albino rats of Wistar strain (breeding of the Institute of Phys-
iology, Academy of Sciences of the Czech Republic) were used.
Animals were kept under standard conditions (12/12 h light/dark
cycle, temperature 22 ± 1 ◦ C, humidity 50—60%) with a free access
to food and water. The experimental protocol was approved by Ani-
mal Care and Use Committee of the Institute of Physiology to be in
agreement with Animal Protection Law of the Czech Republic (which
is fully compatible with European Community Council directives
86/609/EEC as well as with NIH Guidelines).
Status epilepticus
Status epilepticus (SE) was elicited in 12- and 25-day-old rats by
pilocarpine (40 mg/kg, i.p.; # P6503, Sigma Chemical Co., St. Louis,
MO) 20—24 h after LiCl (3 mequiv./kg i.p.; # L-0505, Sigma Chemical
Co., St. Louis, MO) pretreatment. Paraldehyde (0.3 ml/kg, i.p.; #
76260, Fluka Chemie AG, Buchs, Switzerland) was injected after
2 h of continuous convulsions to prevent mortality. Younger animals
were returned to their mothers, older ones needed an intensive care
for at least 2 days after SE. Control animals received saline instead
of pilocarpine, other treatments were the same as in SE group.
Surgery
Cortical stimulation and registration electrodes were implanted
under ether anesthesia 3, 6, 9, 13 and/or 26 days after SE. Silver
electrodes were implanted epidurally—–two over the right senso-
rimotor cortical area (AP + 1 and −1; L = 2 mm) for stimulation;
registration electrodes over left sensorimotor cortical area (AP = 0;
L = 2 mm), over left association parietal area (AP = 3; L = 3 mm), and
over left and right visual cortical regions (AP = 6; L = 4 mm). The
coordinates in parentheses are for adult rats; they were recalcu-
lated for immature animals on the basis of bregma-lambda distance.
An indifferent electrode was put into nasal bone, ground electrode
over cerebellum. All silver electrodes were fixed to the skull by
fast curing dental acrylic (Duracrol® Dental, Prague). Implanta-
tion of electrodes lasted less 8—10 min, then ether anesthesia was
interrupted. The experiment started after 1-h rest, rat pups with
immature thermoregulation (up to the end of the third postnatal
week) were on pad heated to 34 ◦ C during this period as well as
during the experiment.
Stimulation procedure
Animals were placed individually into plastic boxes and connected
to the amplifier and stimulator. Every rat had an easy access to food
pellets and water.
Cortical epileptic afterdischarges were elicited by a constant
current stimulator (15-s series of 1-ms biphasic rectangular pulses
with 8-Hz frequency). Intensity was increased stepwise from 0.2 to
15 mA; at least 10-min interval was preserved between stimulations
with individual intensities (Fig. 1). EEG was digitalized at a rate of
500 Hz and saved on the hard-disc (Kaminskij Biomedical Research
Instruments, Prague). It was recorded before, during and 2 min (as
minimum) after stimulation. The behavior of animals was coded into
180
Figure 1 Original recordings of cortical afterdischarges from
the left sensorimotor area of 18-day-old (12 + 6) control rat pup.
Individual traces represent increasing stimulation intensities
(indicated by black numbers). Clear spike-and-wave ADs were
elicited by intensities from 0.6 to 5.0 mA, mixed type of ADs was
induced by 6.0-mA stimulation. Individual recordings from left:
last 2 s of 15-s stimulation (limited by a line) and 40 s after the
end of stimulation. Time mark 2 s, amplitude calibration equals
to 1 mV.
the EEG recording. The type, duration, and thresholds of ADs and
motor phenomena were evaluated. As duration of ADs is concerned,
three calculations were done: total duration without relation to the
type of ADs; duration of spike-and-wave ADs (i.e. from mixed ADs
only a part formed by SW activity was taken); duration of mixed ADs.
Modified Racine’s five-point scale (0: no activity; 1: motor activities
not synchronous with stimuli and/or EEG spikes; 2: head jerks; 3:
clonic forelimb movements; 4: clonic forelimb movements + rearing;
5: clonic forelimb movements + rearing + falling) and symbols were
used for quantification of behavior (Racine, 1972; Mareš et al.,
2002). Rats could be used only once, 7—10 animals formed each
group.
Statistics
Data for thresholds and duration of afterdischarges and intensity
of motor phenomena were statistically evaluated by means of Rank
Sum Mann—Whitney test. Incidence of mixed and recurrent ADs was
evaluated by means of Fisher Exact Test (SigmaStat® SYSTAT). The
level of statistical significance was put on 5%.
Results
Thresholds
P12 group
Developmental changes of thresholds for the first three phe-
nomena (movements during stimulations, spike-and-wave
afterdischarges and clonic seizures) in control rats exhib-
ited an U-shape pattern with the lowest value at the age
of 21 days. This developmental curve was distorted in SE
animals—–there was an increase of threshold values at the
9-day interval after SE (i.e. at the age of 21 days). Both
SE and control rats exhibited the highest thresholds at the
G. Tsenov et al.
Figure 2 Thresholds for cortically induced phenomena
(mean + S.E.M.). From top to bottom: movements during stimu-
lation; spike-and-wave afterdischarges; clonic seizures; mixed
afterdischarges (transition into the limbic type); recurrent
afterdischarges. Left column: P12; right column: P25 group.
Abscissae: intervals after SE in days: 3, 6, 9, 13 and 26; ordi-
nates: threshold current intensity in mA. Control group: white
columns and SE: dashed columns. Asterisks at pairs of columns
denote a significant difference between SE and control animals.
longest interval, i.e. at the age of 38 days. Significantly
lower threshold for movements directly elicited by stimu-
lation was found at 3-day interval after SE in comparison
with the age-matched control group. The same change was
found at 13-day interval (Fig. 2). Threshold current inten-
sities for elicitation of spike-and-wave afterdischarges and
clonic seizures tended to be lower 3, 6 and 13 days after
SE than in corresponding control groups but the level of
significance was not reached.
The incidence of mixed and recurrent ADs was negligible
at short intervals after SE and increased gradually with age
(Table 1). This behavior was especially expressed at 3-day
interval, when mixed type of afterdischarges was recorded
in 4 out of 10 control animals but in none of 7 rats after SE;
and in 7 out of 9 control animals compared with 4 out of 9
rats 6 days after SE. Recurrent ADs were recorded since the
age of 18 days only in control rats (4 out of 9); 3 days later
(i.e. 9 days after SE) recurrent AD appeared in 1 out of 8 SE
rats compared to 5 out of 7 animals in the control group.
Minor changes were observed at longer intervals.
Threshold for mixed type of ADs was higher in SE than in
control groups after 9 and 26 days, no significant differences
Changes of cortical epileptic ADs after SE in rats
Table 1 Incidence of mixed seizures and recurrent after discharges
P12
3 days 6 days 9 days 13 days
Mixed ADs
Control 4/10 7/9 6/7 5/7
SE 0/7 4/9 5/8 8/8
Recurrent ADs
Control 0/10 4/9 5/7 6/7
SE 0/7 0/9 *1/8 7/8
Data are expressed as number of animals with observed phenomena/number animals in group in both rats with SE at P12 (left part) and
at P25 (right part) and in corresponding control groups.
were found at other intervals. Significantly higher thresh-
olds were found for recurrent afterdischarges only at 26-day
interval after SE when compared with controls.
P25 group
No significant changes were found in P25 group as the thresh-
olds of the first three phenomena (movements bound to
stimulation, spike-and-wave ADs, and clonic seizures accom-
panying these ADs) are concerned (Fig. 2). Developmental
changes were only marginal in both SE and control groups.
Incidence of mixed type and recurrent ADs showed that
spread of epileptic activity into limbic system was not con-
siderably influenced (Table 1). Four out of 9 rats 3 days after
SE generated mixed type of afterdischarges and only one
exhibited recurrent ADs in comparison with control group
where all animals had both phenomena. This was the only
difference found; SE and control animals at other intervals
yielded similar results. Significant differences of thresholds
were found for mixed type of ADs: animals 9 days after SE
needed higher current intensities than control rats. Simi-
lar difference was found for threshold of recurrent ADs in
animals 6 days after SE.
Duration
P12 group
Total duration of ADs 3 days after SE was shorter than in
controls if intensities from 3 to 14 mA were used. Duration
of spike-and-wave activity was shorter, too, significant dif-
ferences were the same as for total duration of ADs (Fig. 3).
Opposite changes (longer ADs in SE rats) were registered 3
days later (i.e. 6 days after SE); the level of statistical sig-
nificance was reached with low stimulation intensities (from
1 to 4 mA) for both total duration of ADs and spike-and-wave
ADs. Mixed ADs which appeared in SE group regularly for the
first time at the 9-day interval (i.e. at the age of 21 days) did
not exhibit any significant changes (data not shown). Dura-
tion of either type of ADs was not systematically changed
at longer intervals after SE, only isolated significant differ-
ences were found.
P25 group
Rats in SE P25 group exhibited shorter total as well as
spike-and-wave ADs than control rats not only 3 but also
6 days after SE (Fig. 3). This difference was observed at
181
P25
26 days 3 days 6 days 9 days 13 days 26 days
9/10 7/7 8/8 10/10 10/10 10/10
10/10 4/9 6/8 10/10 10/10 8/8
10/10 7/7 7/8 9/10 10/10 10/10
10/10 *1/9 7/8 8/10 7/8 8/8
many stimulation intensities, especially at the interval of
6 days (from 1.4 to 8 mA, differences in values for 10- and
12-mA stimulation intensities did not reach the level of sta-
tistical significance). Significant differences in duration of
spike-and-wave ADs were again the same as those in total
duration. Mixed type of ADs participated in changes in total
duration especially if higher stimulation intensities were
used (data not shown but see the difference between total
duration and duration of spike-and-wave ADs at higher stim-
ulation intensities in Fig. 3) but the statistical results were
not changed. Again, there were no systematic differences
at longer intervals after SE.
Motor phenomena
Intensity of movements directly elicited by stimulation as
well as of clonic seizures accompanying spike-and-wave type
of ADs were not significantly changed at any interval after
SE in both P12 and P25 groups (data not shown).
Discussion
Individual phenomena evaluated in our experimental
paradigm are generated by different mechanisms. Move-
ments directly elicited by stimulation represent an
activation of the motor system—–sensorimotor cortex is stim-
ulated and movements started always at the opposite side
of the body. Spike-and-wave EEG activity is generated by
thalamo-cortical mechanisms; it was demonstrated for spon-
taneous spike-and-wave episodes in rats (Avanzini et al.,
1992) and it is highly probable that this activity char-
acteristic for the first type of ADs has the same origin.
Clonic seizures accompanying spike-and-wave ADs are again
a reflection of activation of the motor system—–individual
jerks are synchronous with sharp elements in the EEG.
The second type of ADs, elicited only by high intensities
of stimulation current (Mareš et al., 2002) is due to a
spread of epileptic activity into limbic system (EEG as well
as behavioral correlates are identical with those induced
by hippocampal stimulation—–(Swartzwelder et al., 1979;
Bragin et al., 1997)). This transition is realized by means
of polysynaptic connections between thalamus and limbic
structures and mediodorsal thalamic nucleus could be a pri-
mary structure in this transition (Bertram et al., 2001) but
other possibilities have to be taken into account.
182 G. Tsenov et al.

Figure 3 Duration of afterdischarges (mean + S.E.M.) elicited by individual current intensities. Upper box: P12 + 3 and P12 + 6—–P12
groups; lower box: P25 + 3 and P25 + 6—–P25 groups at 3- and 6-day interval, respectively. Abscissae: stimulation current intensities
in mA; ordinates: upper part of each graph denotes total duration of ADs, bottom part: duration of spike-and-wave ADs in seconds.
Other details as in Fig. 2.

The above-mentioned phenomena are differently influ-
enced by SE. Thresholds for movements bound to stimulation
were significantly decreased only in P12 group at the short-
est interval and again 13 days after SE. The acute change is in
agreement with our finding on the response to a series of five
stimuli (frequency potentiation—–Tsenov and Mareš, 2007)
but a decreased threshold at the longer interval is in contra-
diction with changes in single interhemispheric responses,
they are lower in SE rats than in controls at that inter-
val (Mareš et al., 2005). Thresholds for spike-and-wave ADs
and clonic seizures were not changed in either group at
any interval after SE. Again, it is in contrast to a steeper
input—output curve of single responses at long intervals
after SE in P25 group (Mareš et al., 2005). These discrep-
ancies between changes in evoked potentials and epileptic
afterdischarges suggest that their mechanisms of genera-
tion are different—–pure cortical in single evoked responses
(or at least of their initial components—–Grafstein, 1959)
and an involvement of reverberatory thalamo-cortical cir-
cuits in origin of spike-and-wave ADs (Avanzini et al., 1992;
Snead, 1992; Seidenbecher et al., 1998). Unfortunately,
neurotransmitter mechanisms of cortical epileptic ADs are
not known. Our pharmacological data with NMDA receptor
antagonists indicate an important role of these receptors in
adult (Lojková et al., 2006) as well as immature rats (Mareš
and Šlamberová, 2006; data on file). Analysis of a partici-
pation of these as well as other receptors in generation and
maintenance of cortical epileptic afterdischarges represents
further direction of our research.
Changes in transition into the limbic type of ADs and
appearance of recurrent ADs were completely different.
These phenomena were suppressed at short intervals and
significant increase in threshold current intensities were
found at different intervals in both age groups. Their
Changes of cortical epileptic ADs after SE in rats
delayed appearance in P12 group (transition into the lim-
bic type of ADs appeared in more than half of experimental
animals 9 days after SE and recurrent AD even 4 days later
whereas control rats exhibited these phenomena at earlier
age, Table 1) can be interpreted as a developmental delay
due to an insult—–SE. This insult takes place at the moment
of brain growth spurt (Dobbing and Sands, 1971) therefore
extreme activity during SE might interfere with processes
necessary for normal development. Developmental delay
after SE in this age group was found also in body weight as a
simple measure of development (Kubová et al., 2004) as well
as in a complex behavioral process—–acute habituation in the
open field (Kubova et al., in preparation). Another factor to
be taken into account is the damage of the most probable
pathway through mediodorsal thalamic nucleus (Bertram et
al., 2001); marked degeneration of neurons was described
in this nucleus even in rats undergoing SE at P12 (Kubová
et al., 2001, 2002; Druga et al., 2005). The probability of
participation of these two processes is supported by less
expressed changes in P25 group where the connections (not
only through mediodorsal nucleus) are mature and nearly all
control rats exhibit this transition (present results, Mareš et
al., 2002).
Age-related changes in duration of afterdischarges were
found in our experiments. Duration of ADs was significantly
decreased 3 days after SE in both age groups. This decrease
was observed even 6 days after SE in P25 rats, P12 animals
exhibited a significant prolongation of ADs at this inter-
val. This increase in duration is in controversion with an
increase of benzodiazepine receptors in the cerebral cortex
(Rocha et al., 2007) as well as of ␣1 subunit of GABA-
A receptors in granule cells of the dentate gyrus 1 week
after SE in immature rats. This increase is taken for an
adaptive anticonvulsant mechanism (Raol et al., 2006). On
the other hand, an impairment of inhibition mediated by
GABA-B receptors was found in limbic structures (Mangan
and Lothman, 1996); the GABA-B system plays an important
role in arrest of cortical epileptic afterdischarges (Mareš,
2006; Mareš, submitted). The reversal of changes in P12
rats between the third and sixth day after SE with sub-
sequent normalization suggests a possibility of a transient
impairment of GABA-B or other inhibitory system. These
possibilities have to be further analyzed.
Changes in cortical excitability after SE are much less
expressed than changes in hippocampal excitability after
different models of SE in immature rats. In contrast to
papers studying hippocampal excitability (Jensen et al.,
1992, 1998; Lee et al., 1995; Chen et al., 1999; Dube et
al., 2000; Jensen and Baram, 2000; Cilio et al., 2003; Zhang
et al., 2004) data on neocortical excitability are sparse.
They originate either from our (Dóczi et al., 2003; Mareš
et al., 2005; Tsenov and Mareš, 2007) or from Cavalheiro’s
(Sanabria et al., 2002; Silva et al., 2005a,b) laboratory
and they all demonstrate rather subtle changes including
changes in inhibitory and excitatory receptors (Silva et al.,
2005a,b; Rocha et al., 2007). The same is true for mor-
phological damage—–marked damages in limbic structures
and only moderate changes in neocortex (Cavalheiro et al.,
1991; Leite et al., 1996; Mareš et al., 2005; Nairismagi
et al., 2006). Degenerating neurons in cortical motor and
somatosensory areas (which are rather sparse) have a shape
of interneurons meanwhile pyramidal cells remain intact
183
(Mareš et al., 2005). In spite of the fact that morphological
changes in P12 are not so extensive as in P25 or adult group,
marked degeneration of pyramidal neurons was observed in
hippocampus (especially in CA1 field) in rats undergoing SE
at the age of 12 days whereas degenerating interneurons are
not numerous (Druga et al., in preparation). Thus the mor-
phological damage in neocortex and hippocampus is not only
quantitatively but also qualitatively different. All these find-
ings are in agreement with our data on spontaneous seizures
as a late consequence of SE—–all seizures in rats with SE at
PD12 are nonconvulsive and epileptic activity is restricted to
hippocampus (Kubová et al., 2004). Failure of generalization
(or at least of spread into cerebral cortex) of seizure activ-
ity in rats with SE at PD12 is probably not due to changes
in neocortex—–there is only a moderate difference from ani-
mals with SE at PD25 (Mareš et al., 2005) where convulsive
seizures with corresponding electrocorticographic pattern
were recorded (Kubová et al., 2004). It might indicate dif-
ferences in structures mediating connections between old
and new cortical structures. Further analysis will be nec-
essary to understand differences in consequences of SE in
neocortex and limbic system. Participation of serious wors-
ening of the conditions after SE (acute changes in circuitry
responsible for shorter afterdischarges 3 days after SE),
compromising of inhibitory systems and a possible develop-
mental delay remains to be elucidated. Changes of cortical
epileptic afterdischarges found in the present study can
reflect rather a developmental delay than a shift to an easy
epileptogenesis. They might be interpreted that neocorti-
cal damage is not sufficient to form a basis for generation of
epileptic seizures in this brain structure in either age group
studied as well as for an easy spread of epileptic activity
originating in limbic structures into the cerebral cortex in
rats with SE at PD12. The impact of level of brain matura-
tion at the moment of SE induction is reflected also in the
fact that SE induced in adult rats resulted in appearance of
seizures in nearly all animals (Cavalheiro et al., 1991) mean-
while SE at PD25 led to approximately 75% and SE at PD12
to 25% of epileptic rats, respectively (Kubová et al., 2004).
Animals in the present study were not videoEEG monitored
but the relatively long latency of spontaneous seizures after
SE in immature rats (Raol et al., 2006) means that all our
recordings were made during the latent period. Longer inter-
vals after SE and a combination of videoEEG monitoring with
stimulation have to be used for future analysis.
Acknowledgements
This study was supported by grant nos. 309/03/0770 and
304/05/2582 of the Grant Agency of the Czech Republic and
by a research project AV0Z 50110509. The authors would like
to express their thanks to Ms. Irina Necheva for excellent
technical assistance.
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