Seizure-sensitive Gerbils Research Articles

The Role of Gastrodin on Hippocampal Neurons after N-Methyl-D-Aspartate Excitotoxicity and Experimental Temporal Lobe Seizures.

Tian ma (Gastrodia elata, GE) is an ancient Chinese herbal medicine that has been suggested to be effective as an anticonvulsant and analgesic, and to have sedative effects against vertigo, general paralysis, epilepsy and tetanus. The primary active ingredient isolated from GE is termed gastrodin, which is the glucoside of 4-hydroxybenzyl alcohol (4-HBA). Gastrodin can abolish hypoxia-, glutamate- and N-methyl-D-aspartate (NMDA) receptor-induced toxicity in primary culture of rat cortical neurons, and reduces seizure severity in seizure-sensitive gerbils. We evaluated the effect of gastrodin on NMDA excitotoxicity in hippocampal slice cultures (HSCs) with propidium iodide (PI) fluorescence measurement. We also evaluated the effects of gastrodin for treating active in vivo temporal lobe seizures induced by lithium/pilocarpine. Seizure severity, time span to seizure onset, mortality rate and hippocampal histology for survivors were compared. The effect of gastrodin was evaluated for treating in vitro seizures induced by Mg²⁺-free medium in hippocampal slices. Frequencies and amplitudes of epileptiform discharges were compared. The effect of gastrodin on synaptic transmission was evaluated on hippocampal CA1 Schaffer collaterals. Application of 25 μM gastrodin significantly suppressed NMDA excitotoxicity in CA3 but not in CA1 hippocampus and dentate gyrus. Intraventricular gastrodin accelerated seizure onset for 12 min after intraperitoneal pilocarpine injection (P = 0.051). Three of five rats (60%) in the gastrodin group, and three of four (75%) in the dimethyl sulfoxide (DMSO) group died within 3 days after status epilepticus (SE). Gastrodin also failed to inhibit epileptiform discharges in hippocampal slices induced by Mg²⁺-free medium, believed to be NMDA receptor-mediated spontaneous activity. The frequencies of the spontaneous epileptiform discharges were similar under treatments with 25 μM gastrodin, 200 μM gastrodin and DMSO. For the evaluation of gastrodin on synaptic transmission, application of DMSO, 25 μM or 200 μM gastrodin had no significant effect on excitatory postsynaptic potential (EPSP) slopes. Gastrodin at 200 μM decreased paired-pulse facilitation (PPF) from 1.23 ± 0.04 to 1.12 ± 0.04 (P = 0.002). In conclusion, gastrodin failed to suppress in vivo and in vitro seizures in our study. Gastrodin showed no effect on hippocampal Schaffer collateral EPSP. These findings suggest that gastrodin does not interact with ionotropic glutamate receptors to inhibit NMDA receptor-facilitated seizures. However, gastrodin showed protective effects against NMDA toxicity on cultured hippocampal slices. Nevertheless, gastrodin is still a potential neuroprotective agent against NMDA excitotoxicity, with potential benefits for stroke and patients with epilepsy.

Altered expression of adrenocorticotropic hormone in the epileptic gerbil hippocampus following spontaneous seizure

We investigated the temporal alterations of adrenocorticotropic hormone (ACTH) immunoreactivity in the hippocampus after seizure onset. Expression of ACTH was observed within interneurons in the pre-seizure group of seizure sensitive gerbils, whereas its immunoreactivities were rarely detected in seizure resistant gerbil. Three hr after the seizure, ACTH immunoreactivity was significantly increased in interneurons within all hippocampal regions. On the basis of their localization and morphology through immunofluorescence staining, these cells were identified as GABAA α1-containing interneurons. At the 12 hr postictal period, ACTH expression in these regions was down-regulated, in a similar manner to the pre-seizure group of gerbils. These findings support the increase in ACTH synthesis that contributes to a reduction of corticotrophin-releasing factor via the negative feedback system which in turn provides an opportunity to enhance the excitability of GABAergic interneurons. Therefore, ACTH may play an important role in the reduction of excitotoxicity in all hippocampal regions. [BMB Reports 2013; 46(2): 80-85]

The changes of ERG channel expression after administration of antiepileptic drugs in the hippocampus of epilepsy gerbil model

Ether-a-go-go (ERG) K + channel is a channel of potassium inward rectification. ERG channelopathy may be a cause of sudden unwanted death. The purpose of our study is to assess the effect of antiepileptic drugs on the expression of ERG K + channel in the hippocampus using seizure resistant (SR) and seizure sensitive (SS) gerbils. As compared to controls, in principal neuron of hippocampus ERG immunoreactivity was significantly decreased after administration of AEDs in SS and SR gerbils. In addition, population spike in response to the second stimulus disappeared, thus population spike amplitude ratio was significantly reduced to zero. These findings indicate that AEDs reduce the expression of ERG channel in the hippocampus of the SR and SS gerbils accompanied by the enhancement of paired-pulse inhibition. In addition, the influence of AEDs on ERG expression in the brain may not be relevant to sudden unexpected death in epilepsy.

Altered PLCβ-1 expression in the gerbil hippocampal complex following spontaneous seizure

Although the phospholipase C (PLC)β-1 isoform is associated with spontaneous seizure and distinctively expressed in the telencephalon, the distribution of PLCβ-1 expression in the epileptic gerbil hippocampus remains controversial. Therefore, we determined whether PLCβ-1 is associated with spontaneous seizure in an animal model of genetic epilepsy. In the present study, PLCβ-1 immunoreactivity was down-regulated in seizure-sensitive (SS) gerbils more than in seizure-resistant (SR) gerbils. The expression of PLCβ-1 within calretinin (CR)- positive neurons was rarely detected within the dentate hilar region of SS gerbils. PLCβ-1 immunoreactivity in the hippocampus was significantly elevated as compared to that in pre-seizure SS gerbil 3 h post-ictal. These findings suggest that alterations in PLCβ-1 immunoreactivity in the SS gerbil hippocampus may be closely related to the epileptic state of the gerbil brain and transiently elevated PLCβ-1 protein levels following seizure episodes. Such alterations may be compensatory responses in the SS gerbil hippocampus.

Alteration in NCX-3 immunoreactivity within the gerbil hippocampus following spontaneous seizures

Although NCX-3 is highly expressed in the brain, the distribution of NCX-3 in the epileptic hippocampus is still controversial. Therefore, to assess the distribution and pattern of NCX-3 expression in epileptic hippocampus, we performed a comparative analysis of NCX-3 immunoreactivities in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. In SR gerbils, NCX-3 immunoreactivity was higher than pre-seizure SS gerbils, particularly in the pavalbumin (PV)-positive interneurons. Three h post-ictal, NCX-3 immunoreactivity in the SS gerbil hippocampus was markedly elevated to the level of SR gerbils. Six h post-ictal, the expression of NCX-3 was reduced to the level of pre-seizure SS gerbils. Therefore, the results of the present study suggest that down-regulation of NCX-3 expression in the SS gerbil hippocampus may be involved in the hyperexcitability of SS gerbils due to an imbalance of intracellular Na(+)/Ca(2+) homeostasis and Ca(2+) concentration.

Down-regulation of delayed rectifier K + channels in the hippocampus of seizure sensitive gerbils

In order to confirm the species-specific distribution of voltage-gated K + (Kv) channels and the definitive relationship between their immunoreactivities and seizure activity, we investigated Kv2.x, Kv3.x and Kv4.x channel immunoreactivities in the hippocampi of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. There was no difference in Kv2.1, Kv3.4, Kv4.2 and Kv4.3 immunoreactivity in the hippocampus between SR and SS gerbils. In comparison to SR gerbils, Kv3.1b immunoreactivity in neurons was significantly lower in SS gerbils instead Kv3.1b-immunoreactive astrocytes were clearly observed in SS gerbils ( p < 0.05). Kv3.2 immunoreactivity was also significantly lower in neurons of SS gerbils than in those of SR gerbils ( p < 0.05). Considering the findings of our previous study, these findings suggest that delayed rectifier K + channels (Kv1.1, Kv1.2, Kv1.5, Kv1.6, Kv2.1 and Kv3.1-2), not A-type K + channels (Kv1.4, Kv3.4 and Kv4.x), may be down-regulated in the SS gerbil hippocampus, as compared to SR gerbils.

Voltage-gated Na+ channel II immunoreactivity is selectively up-regulated in hippocampal interneurons of seizure sensitive gerbils

In the present study, we investigated the distribution of voltage-gated Na(+) channels (VGSCs) in the normal and epileptic hippocampus of gerbils (a genetic epilepsy model) in order to confirm the relationship between VGSC and seizure activity in these animals. There was no difference of VGSC I immunoreactivity in the hippocampus between seizure-resistant (SR) and seizure sensitive (SS) gerbils. VGSC II immunoreactivity was rarely detected in the perikarya of principal neurons and interneurons in the SR gerbil hippocampus. However, in the SS gerbil hippocampus, VGSC II immunoreactivity was densely observed in the somata of interneurons located in the stratum radiatum and stratum lacunosum-moleculare. Double immunofluorescent study showed immunoreactivity for calretinin (approximately 80% in VGSC II-positive neurons) or calbindin D-28k (approximately 20% in VGSC II-positive neurons) in VGSC II-immunoreactive neurons. VGSC II-immunoreactive neurons did not show parvalbumin immunoreactivity. These findings suggest that seizure activity in SS gerbils may be related to the selective hyperactivation of interneurons in stratum lacunosum-moleculare via the up-regulation of VGSC II expression, which leads to the disinhibition of CA1 pyramidal cells.

Differential Immunoreactivity of Microglial and Astrocytic Marker Protein in the Hippocampus of the Seizure Resistant and Sensitive Gerbils

In the present study, we compared differences in ionized calcium-binding adapter molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) immunoreactivities for microglia and astrocytes, respectively, in the hippocampus of the seizure-resistant (SR) and seizure-sensitive (SS) gerbils. The density of Iba-1 immunoreactive microglia in the hippocampal CA1 region (CA1) and dentate gyrus (DG) of the SS gerbil was higher than that in the SR gerbil, and many Iba-1 immunoreactive microglia in the SS gerbil were hypertrophied in morphology. In contrast, we could not find significant difference in the density of GFAP immunoreactive astrocytes between the SR and SS gerbils. This result indicates that Iba-1 immunoreactive microglia in CA1 and DG of the SS gerbil are activated compared to those in the SR gerbil.

Reduced Immunoreactivity of Tyrosine Hydroxylase in the Hypothalamic Paraventricular Nucleus of the Seizure Sensitive Gerbil

We compared the immunoreactivity and numbers of tyrosine hydroxylase (TH) immunoreactive neurons and neuropil in the paraventricular nucleus (PVN) of the hypothalamus between the seizure sensitive (SS) and seizure resistant (SR) gerbils. The distributional pattern of TH immunoreactivity was similar in both groups: TH immunoreactivity was seen mainly in magnocellular neurons of the PVN. However, total TH immunoreactivity in the neurons and neuropil in the SS gerbils was significantly lower than that in the SR gerbils. In addition, the number of TH immunoreactive neurons in the SS gerbils was also much lower than those in the SR gerbils. These results indicate that SS gerbils have a low TH immunoreactivity in the hypothalamic PVN compared with that in SR gerbils.

Up-regulated astroglial TWIK-related acid-sensitive K + channel-1 (TASK-1) in the hippocampus of seizure-sensitive gerbils: A target of anti-epileptic drugs

In order to identify the modulation of TASK (TWIK-related Acid-Sensitive K +) channel expressions in epilepsy, we conducted a comparative analysis of TASK channel immunoreactivities in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. There was no difference of the TASK-1 and TASK-2 channel expressions in the hippocampi of young SR and SS gerbils (1–2 months old). In adult SS gerbil hippocampus, TASK-1 immunoreactivity in astrocytes was higher than that in adult SR gerbil hippocampus. After seizures, TASK-1 immunoreactivity was significantly down-regulated in astrocytes of the SS gerbil hippocampus. In addition, various anti-epileptic drugs selectively affect TASK-1 immunoreactivity in astrocytes of the SS gerbil hippocampus. Gabapentin, lamotrigine, topiramate and valproic acid reduced the number of TASK-1 + astrocytes in the hippocampus to 10–25% of that in saline-treated SS adult gerbils, whereas carbamazepine and vigabatrin decreased to ∼ 50%. Therefore, the present study demonstrates that up-regulated TASK-1 immunoreactivity in astrocytes may be involved in the seizure activity of SS adult gerbils and suggests that the astroglial TASK-1 channel may be a target for epilepsy therapeutics.

Parvalbumin immunoreactivity and protein content alter in the hippocampus after adrenalectomy in seizure sensitive gerbils

Objectives: Neurons containing parvalbumin (PV), a calcium-binding protein, in the hippocampus, play an important role in hippocampal excitability in epilepsy. In this study, we examined temporal and spatial changes of PV immunoreactivity and protein content in the hippocampus after adrenalectomy (ADX) in seizure sensitive (SS) gerbils, which are hereditarily seizure-prone.Methods: PV distribution and change in SS gerbils after ADX were examined in the hippocampal CA1 region and in the dentate gyrus (DG) using immunohistochemistry and Western blot analysis.Results: PV immunoreactivity in sham-operated SS gerbils was detected in many CA1 pyramidal cells. Three hours after ADX, PV immunoreactivity significantly decreased in CA1 pyramidal cells and thereafter PV immunoreactivity began to increase by 4 days after ADX. Four days after ADX, PV immunoreactivity was significantly higher than that in the sham-operated SS gerbils. In the DG of sham-operated SS gerbils, PV immunoreactivity was mainly detected in polymorphic cells. Three hours after ADX, PV immunoreactivity in the DG significantly decreased in the polymorphic layer. Thereafter, PV-immunoreactive neurons decreased with time after ADX. Western blot analysis showed that change in PV protein content was similar to immunohistochemical data after ADX in SS gerbils.Conclusion: Our results indicate that PV is changed in hippocampus after ADX and PV may be associated with the regulation of seizure activity.

Seizure activity selectively reduces 5-HT 1A receptor immunoreactivity in CA1 interneurons in the hippocampus of seizure-prone gerbils

Since the correlation between the serotonin (5-hydroxytryptamine, 5-HT) system and seizure activity remains to be clarified, we investigated the 5-HT system in the hippocampus of seizure-resistant (SR) and seizure-sensitive (SS) gerbils. There was no difference of the 5-HT system in the hippocampi of young animals (predisposed and juvenile gerbils) in both SR and SS gerbils. 5-HT immunoreactivity in the dorsal raphe nucleus and the median raphe nucleus was also similarly detected in both animal groups. As compared to SR adult gerbils, only 5-HT 1A receptor immunoreactivity was selectively reduced in CA1 interneurons within SS adult gerbils. (±)-8-Hydroxy-2-(di- n-propylamino)tetralin (a 5-HT 1A receptor agonist, 1 and 2 mg/kg) markedly reduced paired-pulse inhibition in the CA1 region of SS adult gerbils only. These findings suggest that the selective reduction in 5-HT 1A receptor expression on CA1 interneurons of SS adult gerbil may not be developmental defects, but be an acquired compensatory change induced by repeated seizure activity.

Seizure activity affects neuroglial Kv1 channel immunoreactivities in the gerbil hippocampus

In order to confirm the species-specific distribution of voltage-gated K + (Kv) channels and the definitive relationship between their immunoreactivities and seizure activity, we investigated Kv1 channel immunoreactivities in the hippocampus of seizure resistant (SR) and seizure sensitive (SS) gerbils. There was distinct difference of the Kv1 channel subtypes immunoreactivity in the hippocampi in both SR and SS gerbils. Kv1.1, Kv1.2, Kv1.3, Kv1.4, and Kv1.6 immunoreactivities in the SS gerbil hippocampus were lower than that in the SR gerbil hippocampus. However, Kv1 immunoreactivities were obviously presented in astrocyte within the stratum radiatum of the CA1 region of pre-seizure SS gerbil hippocampus. Following seizure-onset, Kv1 immunoreactivities (except Kv1.5) were markedly elevated, whereas their immunoreactivites in astrocytes were down-regulated. Therefore, the present study demonstrates that seizure activity may distinctly affect neuroglial Kv1 immunoreactivities in the gerbil hippocampus.

Altered expression of K +-Cl − cotransporters affects fast paired-pulse inhibition during GABA receptor activation in the gerbil hippocampus

K +-Cl − cotransporter (KCC) plays an important role in maintaining neuronal activity. However, the effect of seizure activity or pharmacological manipulation of GABAergic transmission on KCC expression remains to be clarified. Therefore, the present study was performed to investigate whether seizure activity or GABA receptor agonist treatment changes KCC expression in the gerbil hippocampus. Furthermore, the effect of blockade of KCC on inhibitory transmission in the dentate gyrus was identified following applications of GABA receptor agonists. The distribution of KCC immunoreactivity in the hippocampus was similarly detected between seizure-resistant (SR) and seizure-sensitive (SS) gerbils. Baclofen (a GABA B receptor agonist) treatment markedly increased KCC expression in the gerbil hippocampus. Baclofen treatment significantly reduced paired-pulse inhibition in the dentate gyrus. Furosemide (a KCC inhibitor) treatment amplified the effect of baclofen on paired-pulse responses. In contrast, muscimol (a GABA A receptor agonist) treatment reduced KCC expression. Enhanced paired-pulse inhibition by muscimol treatment was not affected by furosemide treatment. These findings suggest that seizure activity in the gerbil may not affect KCC expression in the hippocampus. In addition, altered KCC immunoreactivity induced by baclofen or muscimol may play an important role in maintaining or regulating inhibitory transmission during GABA receptor activation.

Differential effects of vigabatrin and zonisamide on the neuropeptide Y system in the hippocampus of seizure prone gerbil

Changed neuropeptide Y (NPY) system in the hippocampus has been reported in various experimental epileptic models. However, there have been little data concerning the alteration in the NPY system in the epileptic hippocampus following treatment of anti-epileptic drugs (AEDs). In the present study, therefore, we performed analyses of effects of vigabatrin (VGB) and zonisamide (ZNS) treatment on the NPY system in the hippocampus of the seizure sensitive (SS) gerbils. In SS gerbil, NPY immunoreactivity in the hippocampus was lower than that in seizure resistant gerbil. Following VGB treatment, the number of NPY immunoreactive neurons and NPY mRNA expression were increased in the hilus and the hippocampus proper. In contrast, ZNS treatment markedly elevated only the density of NPY immunoreactive fibers in the dentate gyrus, not in the hippocampus proper, as compared with saline-treated animals. These patterns were observed in the dose-dependent manners. These findings suggest that AEDs treatments may distinctly affect the NPY system in the SS gerbil hippocampus.

Valproic acid reduces enhanced vesicular glutamate transporter immunoreactivities in the dentate gyrus of the seizure prone gerbil

To elucidate the relationship between glutamatergic current and vesicular glutamate transporter (VGLUT) expressions, we performed the comparative analyses of evoked potentials and VGLUT immunoreactivities in the dentate gyrus, and its response to antiepileptic drug treatments in a gerbil model. The EPSP slope that could be evoked in seizure sensitive (SS) gerbils was significantly greater than in seizure resistant (SR) gerbils. There was also a strong trend towards the larger population spike amplitude in SS gerbils. In addition, VGLUT immunoreactivities were markedly enhanced in the dentate gyrus of SS gerbils, as compared with the SR gerbils. Following valproic acid (VPA, 30 mg/kg), the population spike amplitude and the EPSP slope in response to the stimulus were markedly reduced in the dentate gyri both of SR and of SS gerbils, although this dosage of VPA had no effect in low stimulus currents in SS gerbils. Vigabatrin (VGB) and low dosage of VPA treatment did not affect the evoked responses. Similarly, VPA treatment reduced enhanced VGLUT immunoreactivities in the dentate gyrus of SS gerbils, whilst VGB did not. These findings suggest that up-regulation of VGLUT immunoreactivities may be related to the hyperexcitability of granule cells in SS gerbils, and altered VGLUT immunoreactivity in the dentate gyrus may be independent of GABAergic transmission.

Seizure-induced changes of mineralocorticoid and glucocorticoid receptors in the hippocampus in seizure sensitive gerbils

Abnormal corticosteroid hormone levels during stress and resultant mineralocorticoid receptor (MR)/glucocorticoid receptor (GR) imbalance enhance the vulnerability of specific hippocampal neurons. In the present study, we investigated the distribution of MR and GR in seizure resistant (SR) and seizure sensitive (SS) gerbils, and observed the seizure-induced changes of MR and GR in the hippocampus of SS gerbils using immunohistochemistry and western blot analysis. MR and GR immunoreactivities were higher in the SS pre-seizure gerbils than that in SR gerbils. In the SR gerbils, the immunodensity of GR was high compared to that of MR. The changes of MR and GR immunoreactivities were significant in the stratum pyramidale of the hippocampal CA1 region and the infrablade of the dentate gyrus after seizure on-set. MR immunoreactivity in the CA1 region was significantly increased at 12 h after seizure on-set, thereafter MR immunoreactivity was decreased. MR immunoreactivity in the dentate gyrus was decreased time-dependently after seizure on-set. GR immunoreactivity was decreased in the CA1 region and dentate gyrus time-dependently after seizure on-set. At 12 h after seizure on-set, differences in MR and GR immunodensity diminished in the CA1 region and dentate gyrus. This imbalance of MR and GR immunoreactivity in these regions may be associated with seizure generation in the Mongolian gerbil, which is a hereditary seizure model.

Effects of GABAergic transmissions on the immunoreactivities of calcium binding proteins in the gerbil hippocampus

Although reduced calcium binding protein (CBP) immunoreactivities in the epileptic hippocampus have been well established, it has been controversial that these changes may directly indicate neuronal degeneration. In the present study, therefore, we investigated CBP expressions in the gerbil hippocampus following treatment with gamma-aminobutyric acid (GABA) receptor antagonists in order to assess whether altered CBP expressions are the result of either abnormal excitation or indicative of neuronal damage/degeneration. Seizure-sensitive (SS) gerbils showed a loss/decline of CBP immunoreactivities in some hippocampal neurons as compared with seizure-resistant (SR) gerbils. In muscimol (GABA(A) receptor agonist) treated SS gerbils, expression levels of CBP were enhanced as compared with saline-treated SS gerbils. Bicuculline (a GABA(A) receptor antagonist) treatment markedly reduced CBP immunoreactivities in hippocampal neurons of the SR gerbil. Baclofen (a GABA(B) receptor agonist) treatment increased CBP immunoreactivities in the hippocampus of SS gerbils, although its effect was lower than that of muscimol treatment. Moreover, phaclofen (GABA(B) receptor antagonist) treated SR gerbil showed reduction in calbindin D-28K immunoreactivity, not parvalbumin immunoreactivity, in the hippocampus. These findings therefore suggest that reduced CBP immunoreactivities may be the consequence of abnormal discharge caused by loss of GABAergic inhibition rather than an indication of the neuronal damage/degeneration.

Elevated substance P (NK-1) receptor immunoreactivity in the cerebellum of seizure prone gerbil

In the present study, we performed a comparative analysis of the distribution of substance P (SP) receptor (NK-1) immunoreactivity in order to determine the characteristics of the SP system in the cerebelli of rat and gerbils. In the rat cerebellar cortex, only a few Purkinje cells exhibited weak NK-1 receptor immunoreactivity. Similar to the case of rat, NK-1 receptor immunoreactivity in the cerebellar cortex of seizure resistant (SR) gerbils was rarely detected. In contrast, in the cerebellar cortex of seizure sensitive (SS) gerbils, dendrites and cell bodies of Purkinje cell showed strong NK-1 receptor immunoreactivity. Similar to the cerebellar cortex, little NK-1 receptor immunoreactivity in deep cerebellar nuclei was observed in the rat. In SR gerbils, however, deep cerebellar nuclei showed weak NK-1 receptor immunoreactivity. NK-1 receptor immunoreactivity in the deep cerebellar nuclei of SS gerbils was markedly increased, as compared with SR gerbils. Based on the present data, we suggest that the SP system of cerebellar circuit in gerbil are different from rat, and over-expression of NK-1 receptor immunoreactivity in Purkinje cells of SS gerbils may be relevant to Purkinje cell loss induced by seizure activity.

Elevated voltage gated Cl− channel expression enhances fast paired-pulse inhibition in the dentate gyrus of seizure sensitive gerbil

The present study was performed to determine whether the voltage gated Cl- channel (CLC) expression is altered in the hippocampus of seizure sensitive (SS) gerbils, and to identify the strong fast paired-pulse inhibition in the dentate gyrus of SS gerbils is associated with altered CLC expression. In the hippocampal proper and the granule cell layer of the dentate gyrus of the SS gerbils, strong CLC-2 immunoreactivity was detected, as compared with seizure resistant (SR) gerbils. In addition, CLC-3 immunoreactivity was observed in the CA1-3 pyramidal cells, and the granule cell and the molecular layer of the dentate gyrus in the SS gerbils, whereas its immunoreactivity was rarely detected in the SR gerbils. However, CLC-3 immunoreactivity in the mossy fiber was reduced, as compared with SR gerbils. Moreover, infusion of the potential CLC inhibitor (4,4'-diisothiocyanostibene-2,2'-disulfanic acid, DIDS) reduced fast paired-pulse inhibition in the dentate gyrus of SS gerbils, although evoked responses in the dentate gyrus between SR and SS gerbils were similarly detected. These findings suggest that enhancement of CLC expression in the dentate gyrus of SS gerbils may be one of the compensatory responses for reduced GABA(A) receptor-mediated fast postsynaptic inhibitory potentials.