Kainic Acid-induced Limbic Seizures Research Articles

Developmental Hypothyroidism Increases the Expression of Kainate Receptors in the Hippocampus and the Sensitivity to Kainic Acid-Induced Seizures in the Rat

Thyroid hormones are essential for normal brain development, and multiple alterations at behavioral, cognitive, cellular, and molecular levels have been described in animals made hypothyroid during development. Here we analyzed the effect of developmental hypothyroidism in the rat on the sensitivity to kainic acid-induced limbic seizures and the expression of kainate receptors in the hippocampus. Our results show that hypothyroid rats are extremely sensitive to the proconvulsant and neurotoxic effects of kainic acid (KA). Hypothyroid rats entered in status epilepticus at a dose of KA three times lower than that required to reach status epilepticus in control animals. In accordance with this, high levels of glial activation and neuronal loss after low KA dose injections were observed only in the hippocampus of hypothyroid rats. These effects correlated with an increased expression of kainate receptor subunits, excluding GluR5, in the hippocampus of hypothyroid animals. The concentrations of GluR6, GluR7, KAR1, and KAR2 (ionotropic glutamate receptor subunits of the kainic acid subtype) mRNAs were increased between 50 and 250% in hypothyroid animals relative to the values in controls. In agreement with these results, Western blot and immunohistochemical analysis showed a clear increase in the hippocampal content of GluR6/7 proteins in hypothyroid animals.

Directed Evolution of a Novel Adeno-associated Virus (AAV) Vector That Crosses the Seizure-compromised Blood–Brain Barrier (BBB)

DNA shuffling and directed evolution were employed to develop a novel adeno-associated virus (AAV) vector capable of crossing the seizure-compromised blood-brain barrier (BBB) and transducing cells in the brain. Capsid DNA from AAV serotypes 1-6, 8, and 9 were shuffled and recombined to create a library of chimeric AAVs. One day after kainic acid-induced limbic seizure activity in rats, the virus library was infused intravenously (i.v.), and 3 days later, neuron-rich cells were mechanically dissociated from seizure-sensitive brain sites, collected and viral DNA extracted. After three cycles of selection, green fluorescent protein (GFP)-packaged clones were administered directly into brain or i.v. 1 day after kainic acid-induced seizures. Several clones that were effective after intracranial administration did not transduce brain cells after the i.v. administration. However, two clones (32 and 83) transduced the cells after direct brain infusion and after i.v. administration transduced the cells that were localized to the piriform cortex and ventral hippocampus, areas exhibiting a seizure-compromised BBB. No transduction occurred in areas devoid of BBB compromise. Only one parental serotype (AAV8) exhibited a similar expression profile, but the biodistribution of 32 and 83 diverged dramatically from this parental serotype. Thus, novel AAV vectors have been created that can selectively cross the seizure-compromised BBB and transduce cells.

Epileptic Focus Stimulation and Seizure Control in the Rat Model of Kainic Acid-Induced Limbic Seizures

The inhibitory effects of deep brain stimulation (DBS) were investigated in a rat model of kainic acid (KA)-induced limbic status epilepticus. Wistar rats were injected with 1.0 microg KA into the left amygdala after stereotactic implantation of a guide cannula and electrodes. Bipolar rectangular pulses of 0.1 msec duration and 0.1-0.3 mA amplitude were applied intermittently to the left amygdala (10 Hz or 130 Hz), left ventral hippocampus (10 Hz), and left dorsomedial thalamus (130 Hz). Seizure frequency was evaluated by video electroencephalography monitoring and compared to control animals that did not receive DBS. All rats developed limbic status epilepticus 60-90 minutes after KA injection. Seizure frequency was significantly reduced by 10 Hz stimulation of the amygdala and by 130 Hz stimulation of the dorsomedial thalamus. No significant effects were observed with other types of stimulation. Seizure behaviors or duration of seizure were not changed significantly by DBS treatment. DBS of an epileptic focus may attenuate KA-induced limbic seizures, depending on the stimulation sites and parameters.

Cardiac sympathetic nerve activity during kainic acid–induced limbic cortical seizures in rats

We sought to define changes in cardiac sympathetic nerve activity that occur during seizures. We studied kainic acid-induced limbic cortical seizures in urethane-anesthetized rats using cardiac sympathetic nerve, blood pressure, and electrocardiography (ECG) recordings. We studied changes in ventilation rate before and during seizures. Cardiac sympathetic nerve activity was increased during limbic cortical seizures. The modest increases were similar to changes induced by nitroprusside infusion. The normal relation of cardiac sympathetic nerve activity to ventilation rate was lost during seizure activity. Changes in cardiac sympathetic nerve activity caused by changes in ventilation rate became unpredictable, and could be extreme. We conclude that the modest changes in cardiac sympathetic nerve activity contribute to the predominantly parasympathetic effects on the heart during limbic cortical seizures and periods of asphyxia. Further, ventilation rate changes might be associated with large sudden increases or decreases in cardiac sympathetic outflow during seizures.

Anterior pericaudate white matter is important for the secondary generalization of kainic acid-induced limbic seizures in rats

Anterior pericaudate white matter is important for the secondary generalization of kainic acid-induced limbic seizures in rats

Limbic Seizures Induce P-Glycoprotein in Rodent Brain: Functional Implications for Pharmacoresistance

The causes and mechanisms underlying multidrug resistance (MDR) in epilepsy are still elusive and may depend on inadequate drug concentration in crucial brain areas. We studied whether limbic seizures or anticonvulsant drug treatments in rodents enhance the brain expression of the MDR gene (mdr) encoding a permeability glycoprotein (P-gp) involved in MDR to various cancer chemotherapeutic agents. We also investigated whether changes in P-gp levels affect anticonvulsant drug concentrations in the brain.MdrmRNA measured by RT-PCR increased by 85% on average in the mouse hippocampus 3–24 hr after kainic acid-induced limbic seizures, returning to control levels by 72 hr. Treatment with therapeutic doses of phenytoin or carbamazepine for 7 d did not changemdrmRNA expression in the mouse hippocampus 1–72 hr after the last drug administration. Six hours after seizures, the brain/plasma ratio of phenytoin was reduced by 30% and its extracellular concentration estimated by microdialysis was increased by twofold compared with control mice. Knock-out mice (mdr1a/b −/−) lacking P-gp protein showed a 46% increase in phenytoin concentrations in the hippocampus 1 and 4 hr after injection compared with wild-type mice. A significant 23% increase was found in the cerebellum at 1 hr and in the cortex at 4 hr. Carbamazepine concentrations were measurable in the hippocampus at 3 hr inmdr1a/b −/− mice, whereas they were undetectable at the same time interval in wild-type mice. In rats having spontaneous seizures 3 months after electrically induced status epilepticus,mdr1mRNA levels were enhanced by 1.8-fold and fivefold on average in the hippocampus and entorhinal cortex, respectively. Thus, changes in P-gp mRNA levels occur in limbic areas after both acute and chronic epileptic activity. P-gp alterations significantly affect antiepileptic drugs concentrations in the brain, suggesting that seizure-inducedmdrmRNA expression contributes to MDR in epilepsy.

Limbic seizures induce P-glycoprotein in rodent brain: functional implications for pharmacoresistance.

The causes and mechanisms underlying multidrug resistance (MDR) in epilepsy are still elusive and may depend on inadequate drug concentration in crucial brain areas. We studied whether limbic seizures or anticonvulsant drug treatments in rodents enhance the brain expression of the MDR gene (mdr) encoding a permeability glycoprotein (P-gp) involved in MDR to various cancer chemotherapeutic agents. We also investigated whether changes in P-gp levels affect anticonvulsant drug concentrations in the brain. Mdr mRNA measured by RT-PCR increased by 85% on average in the mouse hippocampus 3-24 hr after kainic acid-induced limbic seizures, returning to control levels by 72 hr. Treatment with therapeutic doses of phenytoin or carbamazepine for 7 d did not change mdr mRNA expression in the mouse hippocampus 1-72 hr after the last drug administration. Six hours after seizures, the brain/plasma ratio of phenytoin was reduced by 30% and its extracellular concentration estimated by microdialysis was increased by twofold compared with control mice. Knock-out mice (mdr1a/b -/-) lacking P-gp protein showed a 46% increase in phenytoin concentrations in the hippocampus 1 and 4 hr after injection compared with wild-type mice. A significant 23% increase was found in the cerebellum at 1 hr and in the cortex at 4 hr. Carbamazepine concentrations were measurable in the hippocampus at 3 hr in mdr1a/b -/- mice, whereas they were undetectable at the same time interval in wild-type mice. In rats having spontaneous seizures 3 months after electrically induced status epilepticus, mdr1 mRNA levels were enhanced by 1.8-fold and fivefold on average in the hippocampus and entorhinal cortex, respectively. Thus, changes in P-gp mRNA levels occur in limbic areas after both acute and chronic epileptic activity. P-gp alterations significantly affect antiepileptic drugs concentrations in the brain, suggesting that seizure-induced mdr mRNA expression contributes to MDR in epilepsy.

Regional accumulation of 14C-zonisamide in rat brain during kainic acid-induced limbic seizures.

Zonisamide (ZNS) is an antiepileptic drug developed in Japan. Various experimental studies have investigated the effects of ZNS. However, the mechanism of action of ZNS against limbic seizures and secondary generalization is not well-known. We studied ictal regional accumulation of ZNS in the rat brain during kainic acid (KA)-induced limbic status epilepticus. Fourteen male Wistar rats underwent a stereotactic operation. For recording the electroencephalogram (EEG), electrodes were placed in the left amygdala (LA), left dorsal hippocampus, and over the left sensorimotor cortex. For microinjection, a stainless steel cannula was also inserted into the LA. Seven days after surgery, rats were anesthetized and a catheter was inserted into the femoral vein. The animals were immobilized and allowed to recover from anesthesia for at least two hours. In eight rats, 1.0 microL (1.0 microg) of KA was injected into the LA, and 1.0 microL of phosphate buffer solution was injected into the LA in six control rats. Sixty minutes after injection, 14C-ZNS was administered intravenously, and an autoradiographic study was done. During limbic status epilepticus, only seizures in the sensorimotor cortex were markedly attenuated a few minutes after 14C-ZNS administration. Additionally, high uptake of 14C-ZNS was noted ipsilaterally in the sensorimotor cortex, parietal cortex and thalamus (lateral portion). In control rats, no EEG change was seen, and distribution of 14C-ZNS was rather homogeneous. These results suggested that ZNS suppresses secondary generalization of limbic seizures by a direct effect on the cerebral cortex.

Interruption of supramammillohippocampal afferents prevents the genesis and spread of limbic seizures in the hippocampus via a disinhibition mechanism

In this study we describe the preventive effect of interruption of the supramammillohippocampal afferents on the Fos expression in the forebrain and epileptic discharges in the hippocampal electroencephalogram in rat model of kainic acid-induced limbic seizure. Little was known about the contribution of different degrees of neural activity of hippocampal principal cells to the genesis and spread of limbic seizures in the forebrain structures. Following kainic acid injection to the amygdala with or without concurrent injection of muscimol to the supramammillary nucleus, behavioral changes and electroencephalograms were observed in freely moving rats. The animals were processed for Fos immunocytochemical analysis at several time points. The latest expression of Fos at 2 h was seen in hippocampal CA1–CA3, ventrolateral thalamic nuclei and mediodorsal caudate putamen, while the early Fos expression at 0.5 h was seen in the piriform, entorhinal and other cortices, the thalamic midline nuclei and hypothalamic nuclei. Muscimol injection to the supramammillary nucleus prevented Fos expression in the CA1–CA3 region and reduced that in the forebrain regions with the latest Fos expression, but did not affect Fos expression in other forebrain regions with early Fos expression. This treatment also eliminated epileptic discharges and attenuated all waves in hippocampus. These findings indicate that an acute interruption of the facilitatory hypothalamic afferents by intrasupramammillary injection of muscimol may cause the inactivation of the disinhibition mechanism for hippocampal throughput at the dentate gyrus, resulting in the blockade of the genesis and spread of limbic seizures in the hippocampus.

Antiepileptic effect of nefiracetam on kainic acid-induced limbic seizure in rats

Nefiracetam is being studied as a novel cognition-enhancing agent; however, it has been suggested from studying its chemical structure that it has a potential anticonvulsive effect. We examined the antiepileptic effect of nefiracetam on kainic acid (KA)-induced seizures. KA was infused into the left basolateral amygdaloid nucleus and focal limbic seizures were induced in 43 male Wistar rats. During status epilepticus, 10, 50, 100 or 200 mg/kg of nefiracetam was intravenously injected. Nefiracetam inhibited KA-induced limbic seizures at doses over 100 mg/kg while it had a sedative effect on the animals. In ( 14C) deoxyglucose autoradiographic studies, the propagation of seizure-induced hypermetabolic areas was also suppressed dose-dependently. From the results, it was indicated that nefiracetam has an antiepileptic effect and that its application may suppress seizure propagation. Further study is required, whether this agent is available as a novel anticonvulsant.

Reduction of A1 adenosine receptors in rat hippocampus after kainic acid-induced limbic seizures

In a temporal lobe epilepsy (TLE) model induced by kainic acid (KA), we examined the effect of limbic seizures on A1 adenosine receptor distribution in hippocampus and cortex. By using quantitative autoradiography, we determined a progressive decrease in A1 receptor density in CA1 and CA3 regions of hippocampus, which coincided in time with the degenerating process of hippocampal pyramidal cells. This result indicates that a great amount of A1 receptors are located postsynaptically on pyramidal cell dendrites. No difference in A1 receptor density was observed in the inner compared to the outer molecular layer of dentate gyrus, or in the infrapyramidal band compared to the outer layer of stratum oriens of CA3. This could indicate that the newly sprouted mossy fiber glutamatergic terminals do not contain A1 receptors, thus lacking a restrain in the release of glutamate.

The hippocampal neurons of neuronal apoptosis inhibitory protein 1 (NAIP1)-deleted mice display increased vulnerability to kainic acid-induced injury.

The neuronal apoptosis inhibitory protein (NAIP) is a member of a novel family of inhibitor of apoptosis (IAP) proteins. The IAP genes are highly conserved from baculovirus to metazoans and suppress apoptosis induced by a variety of triggers both in vitro and in vivo. Here we describe the generation and characterization of mice with the targeted deletion of NAIP1. We demonstrate that the NAIP1-deleted mice develop normally. However, the survival of pyramidal neurons in the hippocampus after kainic acid-induced limbic seizures is greatly reduced in the NAIP1 knock-out animals. Thus, although NAIP1 is not necessary for normal development of murine central nervous system, the endogenous NAIP1 is required for neuronal survival in pathological conditions.

Effect of hypothermia on kainic acid-induced limbic seizures: an electroencephalographic and [formula omitted]-deoxyglucose autoradiographic study

The effect of body temperature on kainic acid (KA)-induced limbic seizures was examined in Wistar rats. In rats undergoing limbic seizure induced by 1 μg intra-amygdaloid injection of KA, the post-injection latency of initial ictal discharges in the left amygdala was significantly longer in rats whose body temperature was lowered to 30°C (2.55±0.94 min at 37°C, 13.19±5.70 min at 30°C; p=0.0017). The post-injection latency of initial ictal discharges in the left hippocampus was also significantly longer under the same conditions (23.68±9.96 min at 37°C, 43.85±17.98 min at 30°C; p=0.0253). The number of limbic seizures occurring in the first 2 h post-injection was significantly lower in hypothermic rats (30.0±10.7 at 37°C, 8.71±2.69 at 30°C; p=0.0017), as was the total duration of limbic seizures over the same period (23.61±8.45 min at 37°C, 10.30±4.48 min at 30°C; p=0.0060). Local cerebral glucose utilization (LCGU), measured 2 h post-injection, was significantly lower in hypothermic rats, mainly in the limbic structures. 14 C -deoxyglucose autoradiograms showed decreased radiation density not only in the left amygdala and bilateral hippocampus, but also in the cerebral cortex of hypothermic rats. The results of the present experiment demonstrate that the use of hypothermia, which has been shown to be effective in the treatment of acute cerebral ischemia and brain injury, may also be effective in the treatment of status epileptics.

Behavioral changes and expression of heat shock protein hsp-70 mRNA, brain-derived neurotrophic factor mRNA, and cyclooxygenase-2 mRNA in rat brain following seizures induced by systemic administration of kainic acid

Kainic acid-induced seizures in rats represent an established animal model for human temporal lobe epilepsy. However, it is well-known that behavioral responses to the systemic administration of kainic acid are inconsistent between animals. In this study, we examined the relationship between expression of genes, neuropathological damage, and behavioral changes (seizure intensity and body temperature) in rats after systemic administration of kainic acid. The considerable differences in the response to kainic acid-induced seizures were observed in rats after a single administration of kainic acid (12 mg/kg i.p.). There was no detection of the expression of heat shock protein hsp-70 mRNA and HSP-70 protein in brain of vehicle-treated controls and in animals exhibiting weak behavioral changes (stage 1–2). A moderate expression of hsp-70 mRNA was detected throughout all regions (the pyramidal cell layers of CA1–3 and dentate gyrus) of the hippocampus, the basolateral, lateral, central and medial amygdala, the piriform cortex, and the central medial thalamic nucleus of rats that developed moderate seizures (stage 3–4). Marked expression of hsp-70 mRNA was detected in the all regions (cingulate, parietal, somatosensory, insular, entorhinal, piriform cortices) of cerebral cortex and all regions of hippocampus, and the central medial thalamic nucleus of the rats that developed severe seizures (stage 4–5). In addition, marked HSP-70 immunoreactivity was detected in the pyramidal cell layers of CA1 and CA3 regions of hippocampus, all regions (cingulate, parietal, somatosensory, insular, piriform cortices) of cerebral cortex, and the striatum of rats that developed severe seizures (stage 4–5). Furthermore, a marked expression of cyclooxygenase-2 (COX-2) mRNA and brain-derived neurotrophic factor (BDNF) mRNA levels by kainic acid-induced behavioral seizures (stage 3–4 or stage 4–5) was detected in all hippocampal pyramidal cell layers, granule layers of dentate gyrus, piriform cortex, neocortex, and amygdala. The present study suggest that the behavioral changes (seizure intensity and body temperature) and neuropathological damage after systemic administration of kainic acid are inconsistent between animals, and that these behavioral changes (severity of kainic acid-induced limbic seizures) might be correlated with gene expression of hsp-70 mRNA, COX-2 mRNA, and BDNF mRNA in rat brain.

Increased transcription of glutamate–aspartate transporter (GLAST/GluT-1) mRNA following kainic acid-induced limbic seizure

Expression of mRNA for glutamate–aspartate transporter (GLAST/GluT-1/EAAT1) was studied in the brain of the rat which presented recurrent limbic seizure following systemic administration of kainic acid (KA) by in situ hybridization and Northern blot analysis. The expression of GLAST mRNA was markedly increased after 12 h and peaked after 48 h in animals which demonstrated limbic seizure. The induction of the mRNA were observed in the small non-neuronal cells in the hippocampus, especially around CA3 region and hilus. In contrast, there was no change in GLAST mRNA levels in KA injected seizure-free animals. These findings suggest that GLAST mRNA is induced by seizure and increased extracelluar glutamate levels during seizure may be important for induction of GLAST mRNA.

The anticonvulsant properties of antisense c- fos oligodeoxynucleotides in kainic acid-induced seizures

Evidence has accumulated that the immediate early gene c- fos has important physiological and pharmacological properties in the central nervous system. The role of c- fos in seizures and, in particular, kainic acid-induced seizures, is unclear. It is unknown if c- fos stimulation after kainic acid is a consequence of neuronal activation, or an intrinsic critical component of the metabolic pathways leading to seizure. To elucidate this problem we have pretreated male Wistar rats with antisense c- fos and nonsense c- fos oligodeoxynucleotides 12 h prior to kainic acid 10 mg/kg intraperitoneal. Antisense c- fos inhibited the number of wet dog shakes and the appearance of limbic motor seizures, effects not seen with nonsense or vehicle. The anticonvulsant effects were associated with reduction of both Fos and NGFI-A immunoreactivity and neuroprotection in the hippocampus, thalamus and primary olfactory cortex-amygdaloid region. Four days after antisense c- fos limbic motor seizures were not inhibited, and there was no decrease in Fos or NGFI-A immunoreactivity and no neuroprotection, indicating that the anticonvulsant effects were not secondary to a toxic effect. Sense oligonucleotides had no anticonvulsant effects when given 12 h prior to kainic acid and did not influence immunoreactivity or neuronal survival. In conclusion, these findings suggest a role for c- fos in the generation of kainic acid-induced limbic seizures and neuronal death.

Expression of LIM Protein GenesLmo1, Lmo2,andLmo3in Adult Mouse Hippocampus and Other Forebrain Regions: Differential Regulation by Seizure Activity

The LIM domain is a zinc-binding amino acid motif that characterizes various proteins which function in protein-protein interactions and transcriptional regulation. Expression patterns of several LIM protein genes are compatible with roles in vertebrate CNS development, but little is known about the expression, regulation, or function of LIM proteins in the mature CNS. Lmo1, Lmo2, and Lmo3 are LIM-only genes originally identified as putative oncogenes that have been implicated in the control of cell differentiation and are active during CNS development. Using in situ hybridization for mRNA and immunohistochemical detection of reporter protein expression in transgenic mice, we found that Lmo1, Lmo2, and Lmo3 show individually unique but partially overlapping patterns of expression in several regions of the adult mouse forebrain, including hippocampus, caudate putamen, medial habenula, thalamus, amygdala, olfactory bulb, hypothalamus, and cerebral cortex. In the hippocampal formation, Lmo1, Lmo2, and Lmo3 show different combinatorial patterns of expression levels in CA pyramidal and dentate granule neurons, and Lmo1 is present in topographically restricted subpopulations of astrocytes. Kainic acid-induced limbic seizures differentially regulated Lmo1, Lmo2, and Lmo3 mRNA levels in hippocampal pyramidal and granule neurons, such that Lmo1 mRNA increased, whereas Lmo2 and Lmo3 mRNAs decreased significantly, with maximal changes at 6 hr after seizure onset and return to baseline by 24 hr. These findings show that Lmo1, Lmo2, and Lmo3 continue to be expressed in the adult mammalian CNS in a cell type-specific manner, are differentially regulated by neuronal activity, and may thus be involved in cell phenotype-specific regulatory functions.

Altered expression of NPY-Y 1 receptors in kainic acid induced epilepsy in rats

Kainic acid-induced limbic seizures cause lasting increases in neuropeptide Y (NPY) expression in hippocampal granule cells/mossy fibers. The expression of NPY-Y 1 receptors in these neurons were investigated, using in situ hybridization for Y 1 mRNA and receptor autoradiography with the Y 1-specific ligand [ 125I][Pro 34]PYY. Six hours after kainic acid-induced seizures, Y 1 receptor mRNA levels decreased by 80% in granule cells and concomitantly increased (by 75%) in CA2 pyramidal neurons. Subsequently, persistent decreases in Y 1 mRNA were seen, both in the stratum granulosum and in CA2. Changes in mRNA concentrations were accompanied by a transient, although non-significant, increase in [ 125I][Pro 34]PYY binding in the molecular layer of the dentate gyrus after 4–6 h which was succeeded by a lasting decrease in binding which indicates a persistent down-regulation of Y 1 receptors in hippocampal areas in kainic acid-induced epilepsy.

Increased Mortality of Kainic Acid by Pretreatment with Nicardipine, a Calcium Channel Blocker in Rats.

Ca2+ channel blockers from two different chemical classes, nifedipine and varapamil, were compared in rats for their ability to inhibit the wet-dog shakes behavior and limbic seizures induced by kainic acid which binds to a subtype of glutamate receptors in the brain. Pretreatment for 30 minutes with nicardipine or verapamil suppressed slightly kainic acid-induced wet-dog shakes independent of the dose 1-4mg/kg. In the rats treated with 5mg/kg of nicardipine or verapamil, however, the frequency of wet-dog shakes decreased to 50-60% of controls. In contrast, nicardipine accelerated the onset time and frequency of limbic seizures, resulting in increased mortality by kainic acid in the dose-dependent manner. Occurrence of limbic seizures in verapamil-treated rats was similar to that in the controls and the mortality in this group was smaller than in the nicardipine-treated group. Pretreatment with diazepam alone prevented the kainic acid-induced limbic seizures. However, in the rats pretreated with diazepam and nicardipine simultaneously, anticonvulsive effect of diazepam was not shown and the mortality in this group was the similar to that in the nicardipine pretreated group. Both nicardipine and verapamil did not enhance the picrotoxin-induced convulsions. The mechanism of the apparent enhancement of the mortality of kainic acid by even low doses of nicardipine is not clear. This might be, at least in part, correlated with their different mechanism of channel blocking action.

Transient expression of the mitogen-activated protein kinase phosphatase MKP-1 (3CH134/ERP1) in the rat brain after limbic epilepsy

The immediate early gene-encoded enzyme, MAP kinase phosphatase 1 (MKP-1), is thought to be a key element in controlling cellular signalling pathways activated by MAP kinases. Since MAP kinases have been demonstrated to participate in neuronal stimulus-transcription coupling following seizure activity, the present study investigated the induction of MKP-1 in the rat brain after limbic epilepsy. MKP-1 expression was studied with a polyclonal antiserum by Western blots, immunocytochemistry and immuno-electron microscopy at different time periods between 1 and 24 h after kainic acid-induced limbic seizures. MKP-1 induction was identified in dentate granule cells of the hippocampus but not in pyramidal neurons, furthermore in neurons of the outer layers of the neocortex, as well as in neurons of the lateral nucleus of the bed of the stria terminalis. Immuno-electron microscopy demonstrated that MKP-1 was localized in the neuronal nucleus, where the substrate of MKP-1, activated MAP kinases, are also found. In view of the restricted areas of MKP-1 expression and the widespread areas of altered MAP kinases activity it can be concluded that in the majority of CNS populations other mechanisms than MKP-1 induction are responsible for the shut-off of MAP kinases following seizure activity. MKP-1 may contribute in the specific subpopulations where it is induced to the post-translational control of inducible transcription factors of the fos, jun and myc family.