Systemic Administration Of Kainic Acid Research Articles

Endogenous BDNF Protein Is Increased in Adult Rat Hippocampus after a Kainic Acid Induced Excitotoxic Insult but Exogenous BDNF Is Not Neuroprotective

Systemic administration of the excitotoxin kainic acid to adult rats results in a well defined pattern of loss of the CA1 and CA3 pyramidal neurons of the hippocampus. Prior to this neuronal loss, brain-derived neurotrophic factor (BDNF) mRNA is substantially increased. We show here that BDNF protein is increased after excitotoxic insult in specific areas of the hippocampus, reaching maximal levels 24 h after the insult. BDNF protein levels in the hippocampus increase in direct relation to the severity of seizure. Up to 7 days after injection of kainic acid, levels of full-length Trk B protein were unchanged, whereas levels of truncated TrkB protein were significantly increased by 12 h. To determine whether elevations in BDNF protein levels are potentially beneficial to hippocampal neurons exposed to an excitotoxic stress, we infused exogenous BDNF prior to and during the period of neuronal death caused by kainic acid. We find that administration of high levels of exogenous BDNF does not affect severity of seizure, but does in fact, exacerbate the injury caused by kainic acid, specifically to CA3 pyramidal neurons. Although there was a trend toward sparing of CA1 pyramidal neurons on the side infused with BDNF, this was not significant. In the same paradigm, infusion of exogenous NT-3 had no effect.

A Histochemical Examination of the Staining of Kainate-Induced Neuronal Degeneration by Anionic Dyes

Anionic dyes, notably acid fuchsine, strongly stain the nuclei and cytoplasm of neurons severely damaged by injury or disease. We provide detailed instructions for staining nervous tissue with toluidine blue and acid fuchsine for optimal demonstration of injured neurons. Degeneration was induced in the hippocampus of the mouse by systemic administration of kainic acid, and the resulting acidophilia was investigated using paraffin sections of the Carnoy-or Bouin-fixed brains. The affected cells were bright red with the toluidine blue-acid fuchsine sequence. Their nuclei were stainable also with alkaline Biebrich scarlet and with the 1,2-naphthoquinone-4-sulfonic acid-Ba(OH)2 method; all staining was blocked by benzil but was relatively refractory to deamination by HNO2. These properties indicated an arginine-rich protein. The nuclei were strongly acidophilic in the presence of a high concentration of DNA (strong Feulgen reaction), and acidophilia could not be induced in normal neuronal nuclei by chemical extraction of nucleic acids. The cytoplasmic acidophilia of degenerating hippocampal neurons was due to a protein rich in lysine (extinguished by alkalinity, easily prevented by deamination, and unaffected by benzil). Stainable RNA was absent from the perikarya of the affected cells, but normal neuronal cytoplasm did not become acidophilic after extraction of nucleic acids. We suggest that kainate-induced cell death is preceded by increased production of basic proteins, which become concentrated in the nucleus and perikaryon. Groups of small, darkly staining neurons were seen in the cerebral cortex in control and kainite-treated mice. These shrunken cells were purple with the toluidine blue-acid fuchsine stain, and were attributed to local injury incurred during removal of the unfixed brain.

Type 2 interleukin-1 receptor mRNA is induced by kainic acid in the rat brain

The in situ hybridization technique was used to examine the expression of type 2 interleukin-1 receptor (IL-1R2) mRNA in the rat brain following the systemic injection of kainic acid at a convulsive dose. The expression of IL-1R2 mRNA was not detected in any brain regions of the saline-injected control rats. 8 h after the systemic injection of kainic acid, weak expression of IL-1R2 mRNA was observed in the dentate gyrus and basolateral amygdaloid nucleus. At 12 and 24 h after the injection of kainic acid, IL-1R2 mRNA was markedly induced in various brain regions including the CA1 and CA3 fields of the hippocampus, dentate gyrus, basolateral amygdaloid nucleus, piriform cortex, claustrum, tenia tecta, arcuate hypothalamic nucleus, dorsomedial hypothalamic nucleus, suprachiasmatic nucleus, tuberal magnocellular nucleus and supramammillary nucleus. In these regions, the signals of IL-1R2 mRNA were observed on likely neuronal cells. Around the mediodorsal thalamic nucleus and the paraventricular thalamic nucleus, dispersed intense signals were observed on the non-neuronal cells. In addition, the expression of the mRNA on the venules was observed at 12 h. The strength of signals significantly decreased by 48 h after the injection. These findings revealed the spatiotemporal induction of IL-1R2 mRNA in the rat brain following the systemic administration of kainic acid, which has shown to cause neuronal degeneration, suggesting the pathological roles of IL-1R2 in the brain.

Systemic administration of kainic acid induces selective time dependent decrease in [ 125I]insulin-like growth factor I, [ 125I]insulin-like growth factor II and [ 125I]insulin receptor binding sites in adult rat hippocampal formation

Administration of kainic acid evokes acute seizure in hippocampal pathways that results in a complex sequence of functional and structural alterations resembling human temporal lobe epilepsy. The structural alterations induced by kainic acid include selective loss of neurones in CA1–CA3 subfields and the hilar region of the dentate gyrus followed by sprouting and permanent reorganization of the synaptic connections of the mossy fibre pathways. Although the neuronal degeneration and process of reactive synaptogenesis have been extensively studied, at present little is known about means to prevent pathological conditions leading to kainate-induced cell death. In the present study, to address the role of insulin-like growth factors I and II, and insulin in neuronal survival as well as synaptic reorganization following kainate-induced seizure, the time course alterations of the corresponding receptors were evaluated. Additionally, using histological preparations, the temporal profile of neuronal degeneration and hypertrophy of resident astroglial cells were also studied. [ 125I]Insulin-like growth factor I binding was found to be decreased transiently in almost all regions of the hippocampal formation at 12 h following treatment with kainic acid. The dentate hilar region however, exhibited protracted decreases in [ 125I]insulin-like growth factor I receptor sites throughout (i.e. 30 days) the study. [ 125I]Insulin-like growth factor II receptor binding sites in the hippocampal formation were found to be differentially altered following systemic administration of kainic acid. A significant decrease in [ 125I]insulin-like growth factor II receptor sites was observed in CA1 subfield and the pyramidal cell layer of the Ammon's horn at all time points studied whereas the hilar region and the stratum radiatum did not exhibit alteration at any time. A kainate-induced decrease in [ 125I]insulin receptor binding was noted at all time points in the molecular layer of the dentate gyrus whereas binding in CA1–CA3 subfields and discrete layers of the Ammon's horn was found to be affected only after 12 h of treatment. These results, when analysed with reference to the observed histological changes and established neurotrophic/protective roles of insulin-like growth factors and insulin, suggest possible involvement of these growth factors in the cascade of neurotrophic events that is associated with the reorganization of the hippocampal formation observed following kainate-induced seizures.

Novel Differentially Expressed Genes Induced by Kainic Acid in Hippocampus: Putative Molecular Effectors of Plasticity and Injury

Systemic kainic acid administration in rats induces acute limbicstatus epilepticusand subsequent neuronal degeneration and development of chronic hyperexcitability with similarities to human temporal lobe epilepsy. The mechanisms mediating the responses to kainic acid likely involve transcriptional changes in genes of importance for cellular injury, protection, and plasticity. We have used an arbitrarily primed PCR technique to identify such changes in the rat dentate gyrus. Three previously uncharacterized transcripts were found to be upregulated in the dentate gyrus 4 h following systemic kainic acid.In situhybridization using riboprobes transcribed from the cloned PCR fragments were used to confirm differential expression specifically in dentate granule neurons following seizure. Basal expression for all three transcripts is widespread throughout the rat brain, with the highest levels seen in the hippocampal pyramidal and granule cell layers. The novel sequences do not match any known full-length cDNAs and may belong to novel gene families. However, they all showed high homology to human partial cDNA sequences (ESTs) that are expressed in brain as well as several other tissues. Two additional transcripts identified in this study corroborate earlier findings on differential expression of heat-shock proteins after seizure. The novel transcripts found in this study may be involved in epileptogenesis and neuronal responses to damage following seizure.

Seizure-Induced Damage to Somatostatin-Immunoreactive Neurons in the Rat Hippocampus Is Regulated by Fimbria–Fornix Transection

In both experimental and human temporal lobe epilepsy, seizures cause loss of hilar somatostatin-immunoreactive (SOM-ir) neurons and sprouting of mossy fibers. To investigate whether in rats these alterations are modulated by hippocampal input projections, we transected the fimbria–fornix or the perforant pathway bilaterally 2 days after seizures induced by systemic administration of kainic acid (9 mg/kg, i.p.). Two months later, the number of SOM-ir neurons in the hilus was counted and mossy fiber sprouting in the supragranular area and in the inner molecular layer was analyzed. In seizured rats with sham-operation, 50% of the hilar SOM-ir neurons were left in the septal end of the hippocampus and only 16% remained in the temporal end. In seizured rats with transection of the fimbria–fornix, the number of hilar SOM-ir neurons in the septal end of the hippocampus did not differ from that in controls (98% of SOM-ir neurons left). However, the temporal end was severely damaged (41% of SOM-ir neurons left). In seizured rats with transection of the perforant pathway, 61% of the hilar SOM-ir neurons were left in the septal end and 51% in the temporal end of the hippocampus. Mossy fiber sprouting was evident throughout the septotemporal axis of the hippocampus in all seizured rats. Our results suggest that in the septal end of the hippocampus the severity of neuronal damage in the hilus is modulated by mechanism(s) that are dependent on the afferent pathways entering the hippocampus via the fimbria–fornix. Transection of the fimbria–fornix, however, does not significantly modulate the severity or the target regions of seizure-induced sprouting of mossy fibers.

BFGF and FGFR-3 immunoreactivity in the rat brain following systemic kainic acid administration at convulsant doses: localization of bFGF and FGFR-3 in reactive astrocytes, and FGFR-3 in reactive microglia

Strong bFGF immunoreactivity was observed in reactive astrocytes, as shown by double-labeling immunohistochemistry of bFGF and GFAP, from days 7 up to 30 (last time point examined) following kainic acid (KA) injection at convulsant doses in the adult rat. bFGF was not found in OX-42-positive reactive microglia. A few reactive glia co-localized FGFR-3 and GFAP, whereas the majority of cells expressing FGFR-3 were OX-42-immunoreactive. This was further supported by the observation that only ≈ 10% of reactive glia co-localized bFGF and FGFR-3. These results show that reactive astrocytes are a major source of bFGF during the subacute stages of tissue damage following KA injection and that reactive astrocytes and, most particularly, reactive microglia are putative targets of bFGF through FGFR-3.

Expression of the 27,000 mol. wt heat shock protein following kainic acid-induced status epilepticus in the rat.

Western analysis and immunohistochemistry were used to determine the time-course and the distribution of the 27,000 mol. wt heat shock protein, Hsp27, in rat brain following systemic administration of kainic acid. No Hsp27 immunoreactivity was detected in naive control animals or in rats that failed to develop status epilepticus. Hsp27 immunoreactivity was detected as early as 12 h in the parietal cortex, piriform cortex and the hippocampus of rats that developed status epilepticus. The number of cells expressing Hsp27 and the intensity of Hsp27 immunoreactivity were increased 24 h after kainic acid administration. Hsp27 immunoreactivity was still observed seven days post-kainic acid injection. The morphology of the Hsp27-positive cells and double immunofluorescence against Hsp27 and glial fibrillary acidic protein revealed that Hsp27-positive cells were astrocytes. In addition, the distribution of Hsp27 suggested that astrocytic Hsp27 was dependent on excitation-induced metabolic stress rather than the direct effect of kainic acid on astrocytes.

Absence of excitotoxic neuropathology in microencephalic rats after systemic kainic acid administration

We have s.c. injected with kainic acid (12 mg/kg) normal adult rats as well as rats rendered microencephalic by selectively timed administration of the DNA alkylating agent methylazoxymethanol acetate (MAM) to the mother during pregnancy. Histological examination of the brains revealed that normal animals underwent neurodegeneration in brain regions sensitive to kainic acid excitotoxicity, such as the olfactory cortex and the hippocampus, while no damage was apparent in the same regions of microencephalic rats. Evaluation of the neurotoxic outcome consequent to the excitotoxic stimulation, was quantitatively performed by measuring the levels of appropriate neurochemical markers 15 days after kainic acid injection. In normal animals, this resulted in significant decrease (up to 60% in the olfactory cortex and 30% in the hippocampus) of markers related to glutamatergic and GABAergic neurons, whereas in MAM-treated rats the same markers were not significantly affected, thus demonstrating a substantial protection against the excitotoxic insult in the microencephalic condition.

Absence of excitotoxic neuropathology in microencephalic rats after systemic kainic acid administration

We have s.c. injected with kainic acid (12 mg/kg) normal adult rats as well as rats rendered microencephalic by selectively timed administration of the DNA alkylating agent methylazoxymethanol acetate (MAM) to the mother during pregnancy. Histological examination of the brains revealed that normal animals underwent neurodegeneration in brain regions sensitive to kainic acid excitotoxicity, such as the olfactory cortex and the hippocampus, while no damage was apparent in the same regions of microencephalic rats. Evaluation of the neurotoxic outcome consequent to the excitotoxic stimulation, was quantitatively performed by measuring the levels of appropriate neurochemical markers 15 days after kainic acid injection. In normal animals, this resulted in significant decrease (up to 60% in the olfactory cortex and 30% in the hippocampus) of markers related to glutamatergic and GABAergic neurons, whereas in MAM-treated rats the same markers were not significantly affected, thus demonstrating a substantial protection against the excitotoxic insult in the microencephalic condition.

The inducible 70,000 molecular weight heat shock protein is expressed in the degenerating dentate hilus and piriform cortex after systemic administration of kainic acid in the rat

Using both immunohistochemistry and in situ hybridization, we examined the rat brain for the expression of the inducible 70,000 mol. wt heat shock protein, Hsp70, at 3, 6, 12, and 24 h after systemic administration of kainic acid. In contrast to previous reports, the present study demonstrates that neurons in the regions most susceptible to seizure-induced cell death accumulate both Hsp70 messenger RNA and protein. Neurons in the dentate hilus and piriform cortex contained Hsp70 messenger RNA at 6 h and protein at 12 h. These neurons contained little or no Hsp70 messenger RNA or protein at 24 h when the majority of cells in these areas were pyknotic. Injured neurons in areas such as the parietal cortex, which are less susceptible to seizure-induced cell death, expressed and maintained high levels of Hsp70 messenger RNA and protein at 12 and 24 h. This work suggests that Hsp70 messenger RNA and protein are rapidly and transiently expressed in dying neurons, and contradicts the notion that Hsp70 only accumulates in injured neurons that survive.

Ascorbate attenuates the systemic kainate-induced neurotoxicity in the rat hippocampus

The neuronal damage induced by systemic administration of kainic acid reproduces the cellular and regional pattern of damage produced by repeated seizures, The ability of kainic acid to induce lipid peroxidation, and the ability of free radical inhibitors to prevent ischaemically-induced cell death, has led us to examine the possible role of free radicals in kainate-induced injury, Ascorbic acid was able to reduce kainate-induced damage of the rat hippocampus, measured by means of the gliotic marker ligand [3H]PK1 1195. Ascorbate was significantly effective at doses of 30 mg kg−1 and above, with total protection against kainate at 50 mg kgt-1. Histologically, ascorbate at 50 mg kg−1 was able to prevent kainate-induced neuronal loss in the hippocampal CAI and CA3a cell layers. The antioxidant was also effective when administered simultaneously with, or l h before the kainate. Protection was also obtained by allopurinol, 175 mg kg−1 and by oxypurinol, 40 mg kg−1. Ascorbate did not modify synaptically evoked potentials or long-term potentiation in hippocampal slices, ruling out any blocking activity at glutamate receptors. It is concluded that the neuronal damage produced by systemically administered kainate involves the formation of free radicals.

Ascorbate attenuates the systemic kainate-induced neurotoxicity in the rat hippocampus

The neuronal damage induced by systemic administration of kainic acid reproduces the cellular and regional pattern of damage produced by repeated seizures, The ability of kainic acid to induce lipid peroxidation, and the ability of free radical inhibitors to prevent ischaemically-induced cell death, has led us to examine the possible role of free radicals in kainate-induced injury, Ascorbic acid was able to reduce kainate-induced damage of the rat hippocampus, measured by means of the gliotic marker ligand [ 3H]PK1 1195. Ascorbate was significantly effective at doses of 30 mg kg −1 and above, with total protection against kainate at 50 mg kg t-1. Histologically, ascorbate at 50 mg kg −1 was able to prevent kainate-induced neuronal loss in the hippocampal CAI and CA3a cell layers. The antioxidant was also effective when administered simultaneously with, or l h before the kainate. Protection was also obtained by allopurinol, 175 mg kg −1 and by oxypurinol, 40 mg kg −1. Ascorbate did not modify synaptically evoked potentials or long-term potentiation in hippocampal slices, ruling out any blocking activity at glutamate receptors. It is concluded that the neuronal damage produced by systemically administered kainate involves the formation of free radicals.

Seizure activity causes elevation of endogenous extracellular kynurenic acid in the rat brain

This study was designed to examine the effects of several classic convulsants on the extracellular concentration of the anticonvulsant and neuroprotective brain metabolite kynurenic acid (KYNA) in the rat brain. Drug effects were investigated in vivo, mostly by unilateral microdialysis in the dorsal hippocampus. Systemic administration of pentylenetetrazole (60 mg/kg, SC), pilocarpine (325 mg/kg, SC), bicuculline (6 mg/kg, SC), or kainic acid (10 mg/kg, SC) caused characteristic clonic and/or tonic convulsions. In all seizure paradigms, KYNA levels in the dialysate began to rise within 1 h and gradually reached a plateau approximately 4 h after administration of the convulsants. Peak increases were 1.5-3-fold over basal levels. The duration of the elevation in KYNA levels was significantly prolonged following kainic acid application. In the kainic acid model, extracellular KYNA was also measured and found to be increased in the ventral hippocampus, piriform cortex, and striatum. Moreover, temporary intrahippocampal infusion of the KYN synthesis inhibitor aminooxyacetic acid (1 mM) in the kainic acid- and pentylenetetrazole models attenuated the increase in extracellular KYNA levels, demonstrating that de novo production of KYNA in the brain accounts for the seizure-induced KYNA overflow. A separate group of animals received a unilateral intrahippocampal injection of the endogenous convulsant excitotoxin quinolinic acid (120 nmol) and showed long-lasting (> 24 h) bilateral increases in extracellular KYNA levels. Taken together, these data indicate that an increase in extracellular KYNA may constitute a common occurrence in response to seizures and that KYNA elevations may signify the brain's attempt to counteract seizure activity.

Nicotine blocks kainic acid-induced wet dog shakes in rats.

Systemic or central administration of kainic acid (KA) in rats results in the expression of wet dog shakes (WDS) followed by motoric seizures and convulsions, which are associated with limbic neurotoxicity. Although a number of neurotransmitter systems are thought to be involved with this KA-induced syndrome, little is known about the possible influence of cholinergic nicotinic receptor modulation. In the study presented here, we pretreated rats with saline or 0.5 mg/kg nicotine base followed 15 minutes later by 12.0 mg/kg KA and then observed the incidence of WDS between 45 and 120 minutes post-KA injection. Rats pretreated with nicotine exhibited significantly less WDS than those pretreated with saline (p < .001). Whereas the mechanism for this nicotine effect is currently not known, future experiments will look at dose-response relationships, the role of nicotine receptors, and possible neuroprotective potential of nicotine in this KA-induced syndrome.

Temporalspatial patterns of expression of metallothionein-I and -III and other stress related genes in rat brain after kainic acid-induced seizures

Kainic acid-induced seizures in the rat brain cause severe brain damage that is thought to result, in part, from oxidative stress. In this study, we examine the consequences of systemic administration of kainic acid on expression of several genes that encode proteins thought to play roles in protection from oxidative stress, including metallothionein-1, and -III. Kainic acid causes an increase in metallothionein-I and heme oxygenase-1 mRNAs, as well as an increase in c-fos, heat shock protein-70, and interleukin-1 β mRNAs. The induction of these mRNAs is seizure dependent, and is greater in brain areas with extensive damage (e.g. piriform cortex) than in areas with minimal damage (e.g. frontal cortex and cerebellum). In contrast, little or no change in mRNA for metallothionein-III, manganese superoxide dismutase, copper-zinc superoxide dismutase, glutathione-s-transferase ya subunit or glutathione peroxidase occur. The prolonged and robust concordant induction of the metallothionein-I and heme oxygenase-1 genes may reflect the oxidative stress produced by kainic acid-induced seizures. In addition, the induction of inter-leukin-1β gene expression suggests an inflammatory response in brain regions damaged by kainic acid-induced seizures. Delineating the regulation of genes associated with oxidative and inflammatory responses can contribute to a fuller understanding of seizures and associated brain damage.

Effects of the systemic administration of kainic acid and NMDA on exploratory activity in rats

In spite of growing evidence for the involvement of the glutamatergic system in mammal's locomotion, studies on behavioural effects induced by the systemic administration of excitatory amino acids not associated to convulsions are lacking. In the present work, the effect of one single systemic administration of kainic acid (KA) (9 mg/kg, IP) or NMDA (100 mg/kg, IP) on exploratory activity in the rat during 6 consecutive days was studied. Separation of exploratory activity in fast (FM) and slow movements (SM) and rearings (R), together with the analysis of those variables during both the light and dark periods of the light-dark cycle, allowed finding specific drug-induced effects. KA produced an acute short-lasting increase in exploratory activity, only significant for FM. On the other hand, NMDA produced an acute short-lasting depressant effect on FM, SM, and R, followed during the next 2 days by a long-lasting increase in exploratory activity, only significant for FM during the dark period. These results underline the importance of using repeated testing during both light and dark periods of the light-dark cycle when analyzing drug-induced changes on exploratory activity.

Induction of c‐fos and reduction of dynorphin in dentate granule cells of a rat model of epilepsy produced by systemic administration of kainic acid: An immunohistochemical study

Induction of <i>c‐fos</i> and reduction of dynorphin in dentate granule cells of a rat model of epilepsy produced by systemic administration of kainic acid: An immunohistochemical study

Differential susceptibility to seizures induced by systemic kainic acid treatment in mature DBA/2J and C57BL/6J mice.

Mature DBA/2J (D2) and C57BL/6J (B6) mice aged 9-10 weeks were studied to determine susceptibility to behavioral seizures induced by kainic acid (KA) and the possible influence exerted by differences in metabolism and blood-brain barrier (BBB) transport. Mice were observed for 4 h after subcutaneous (s.c.) KA injection. Behavioral seizure parameters included latency to first seizure (clonus), latency to tonic/clonic seizure, and latency to status epilepticus (SE). At a KA dose of 25 mg/kg, 80% of D2 mice exhibited tonic/clonic seizures, whereas all B6 mice remained seizure-free. At 30 mg/kg, tonic/clonic seizures were observed in 100% of D2 mice and 25% of B6 mice. Of D2 mice exhibiting at least one clonic seizure in response to KA at a dose of 25 mg/kg, 50% entered SE and eventually died. Administration of [3H]KA (6.6 x 10(6) dpm) at doses of 25 mg/kg (convulsive) or 11.1 micrograms (nonconvulsive) to mice of both strains resulted in similar levels of radioactivity in cortex, hippocampus, and cerebellum 30 and 60 min after injection. Bioconversion of [3H]KA to a radiolabeled brain metabolite in vivo could not be documented in mice from either strain. Results confirm previously reported differences between D2 and B6 mice in their relative susceptibility to seizures induced by systemic KA administration and suggest that these differences are not related to strain-specific variation in metabolism or BBB transport of KA. Further studies of these two strains of mice may be useful for investigating genetic influences upon seizure susceptibility.

Kainic acid-induced heat shock protein-70, mRNA and protein expression is inhibited by MK-801 in certain rat brain regions.

The regional expression of inducible 72 kDa heat shock protein (HSP-70), HSP-70 mRNA and the neuropathological outcome of their expression were examined in the rat brain following systemic administration of kainic acid (9 mg/kg), and also after pretreatment with the non-competitive N-methyl-D-aspartate antagonist MK-801 (1 mg/kg). Five hours after administration of kainic acid alone, dense expression of HSP-70 mRNA was found within the limbic system, mainly in the hippocampus, piriform and entorhinal cortices, amygdaloid complex, thalamic nuclei, subiculum and in other cortical areas in rats that had shown convulsive behaviour. At 24 h, HSP-70 immunoreactivity was seen in most areas previously expressing HSP-70 mRNA, except the piriform and entorhinal cortices and several ventral nuclei of the amygdaloid complex. Histopathological examination at 24 h revealed marked cell loss in these latter regions and less severe histopathological changes in other areas of the limbic system in brains of convulsive rats. No alterations were apparent in non-convulsive rats. The percentage of rats showing convulsive behaviour with kainic acid was reduced from 74 to 4% following pretreatment with MK-801. In addition, MK-801 inhibited the kainic acid-induced expression of HSP-70 mRNA and protein in certain brain regions, notably the cortex, the pyramidal cell layer of CA1, and discrete thalamic nuclei. However, HSP-70 mRNA induction was sustained in the pyramidal cell layer of CA3, the amygdaloid complex and the subiculum, despite the fact that none of these rats convulsed. MK-801 prevented necrosis in all rats examined except the single rat that had shown convulsive behaviour. These results show that early regional expression of inducible HSP-70 mRNA allows the visualization of regions affected by kainic acid and maps regions inhibited by MK-801. In addition, the results identify brain regions putatively involved in the manifestation of limbic convulsions. Furthermore, these data illustrate that the induction of HSP-70 mRNA is not predictive of cell death or survival.