Hippocampal damage after intra-amygdala kainic acid-induced status epilepticus and seizure preconditioning-mediated neuroprotection in SJL mice
ABSTRACT Exposure of the brain to a stressful stimulus that is sub-threshold for permanent injury can temporarily protect against cell death during a subsequent and otherwise damaging insult. One or more brief, non-harmful seizure episode(s) (seizure preconditioning) can dramatically reduce hippocampal damage when given prior to status epilepticus (epileptic tolerance). We recently reported that status epilepticus-induced hippocampal damage in C57BL/6 mice could be reduced by approximately 50% when preceded 24h earlier by a brief, non-injurious generalized seizure induced by 15mg/kg systemic kainic acid (KA). Since other mouse strains might display different vulnerability to either seizure preconditioning or status epilepticus, we investigated whether epileptic tolerance could be acquired in another strain. SJL mice, reported to display greater seizure sensitivity to systemic KA, received intra-amygdala microinjection of KA to trigger status epilepticus. Intracerebral recordings confirmed evoked seizures involved the ipsilateral hippocampus. Status epilepticus produced hippocampal damage which mainly affected the ipsilateral CA3 and hilus; a pattern similar to C57BL/6 mice. The damage extended through the full rostro-caudal extent of the hippocampal formation. Seizure preconditioning using 20mg/kg systemic KA, but not 15mg/kg, significantly reduced hippocampal damage after status epilepticus by 37% in the dorsal hippocampus and by 65% in the ventral hippocampus. These studies suggest status epilepticus induced by intra-amygdala KA in SJL mice models aspects of the pathophysiology of human mesial temporal sclerosis. Moreover, seizure preconditioning effectively produces neuroprotection in SJL mice, further establishing epileptic tolerance as a conserved endogenous neuroprotection paradigm.
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ABSTRACT: Prolonged seizures (status epilepticus) produce pathophysiological changes in the hippocampus that are associated with large-scale, wide-ranging changes in gene expression. Epileptic tolerance is an endogenous program of cell protection that can be activated in the brain by previous exposure to a non-harmful seizure episode before status epilepticus. A major transcriptional feature of tolerance is gene downregulation. Here, through methylation analysis of 34,143 discrete loci representing all annotated CpG islands and promoter regions in the mouse genome, we report the genome-wide DNA methylation changes in the hippocampus after status epilepticus and epileptic tolerance in adult mice. A total of 321 genes showed altered DNA methylation after status epilepticus alone or status epilepticus that followed seizure preconditioning, with >90% of the promoters of these genes undergoing hypomethylation. These profiles included genes not previously associated with epilepsy, such as the polycomb gene Phc2. Differential methylation events generally occurred throughout the genome without bias for a particular chromosomal region, with the exception of a small region of chromosome 4, which was significantly overrepresented with genes hypomethylated after status epilepticus. Surprisingly, only few genes displayed differential hypermethylation in epileptic tolerance. Nevertheless, gene ontology analysis emphasized the majority of differential methylation events between the groups occurred in genes associated with nuclear functions, such as DNA binding and transcriptional regulation. The present study reports select, genome-wide DNA methylation changes after status epilepticus and in epileptic tolerance, which may contribute to regulating the gene expression environment of the seizure-damaged hippocampus.The Journal of Neuroscience : The Official Journal of the Society for Neuroscience 02/2012; 32(5):1577-88. DOI:10.1523/JNEUROSCI.5180-11.2012 · 6.75 Impact Factor
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ABSTRACT: Prolonged seizures (status epilepticus) can activate apoptosis-associated signaling pathways. The extent to which such pathways contribute to cell death might depend on the insult intensity, whereby the programmed or apoptotic cell death component is reduced when seizures are more severe or protracted. We recently showed that mice lacking the pro-apoptotic Bcl-2 homology domain 3-only protein Puma (Bbc3) were potently protected against damage caused by status epilepticus. In the present study we examined whether Puma deficiency was protective when the seizure episode was more severe. Intra-amygdala microinjection of 1 microg kainic acid (KA) into C57BL/6 mice triggered status epilepticus that lasted about twice as long as with 0.3 microg KA prior to lorazepam termination. Hippocampal damage was also significantly greater in the higher-dose group. Over 80% of degenerating neurons after seizures were positive for DNA fragmentation assessed by terminal deoxynucleotidyl dUTP nick end labeling (TUNEL). Microscopic analysis of neuronal nuclear morphology in TUNEL-positive cells revealed the proportion displaying large rounded clumps of condensed chromatin was approximately 50% lower in the high-dose versus low-dose KA group. Nevertheless, compared to heterozygous and wild-type mice subject to status epilepticus by high-dose KA, neuronal death was reduced by approximately 50% in the hippocampus of Puma-deficient mice. These data suggest aspects of the apoptotic component of seizure-induced neuronal death are insult duration- or severity-dependent. Moreover, they provide further genetic evidence that seizure-induced neuronal death is preventable by targeting so-called apoptosis-associated signaling pathways and Puma loss likely disrupts caspase-independent or non-apoptotic seizure-induced neuronal death.Neuroscience 03/2010; 168(2):443-50. DOI:10.1016/j.neuroscience.2010.03.057 · 3.33 Impact Factor
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ABSTRACT: One or more brief seizures can serve to activate endogenous protective programmes which render brain regions temporarily less susceptible to damage following an otherwise harmful episode of status epilepticus (a prolonged seizure). Epileptic tolerance has been demonstrated using a variety of seizure preconditioning paradigms, including electroconvulsive shocks and low doses of excitotoxins such as kainic acid. The cell and molecular mechanisms underlying the protection are not fully understood but proposed mediators include the transcription factor NfκB, altered ion channel expression, upregulation of growth factors and other protective genes, and suppression of pro-apoptotic Bcl-2 family proteins. Application of microarrays to profile the transcriptome of seizure-preconditioning and tolerance has provided further insights, including roles for chromatin remodeling and evidence that preconditioning generates an anti-excitotoxicity phenotype by reprogramming the transcriptional response to status epilepticus. This review summarizes the various animal models of epileptic tolerance, reviews the key effector(s) and the utility of this experimental paradigm for identifying novel targets for neuroprotection and anti-epileptogenesis.International Journal of Physiology, Pathophysiology and Pharmacology 01/2009; 1(2):180-191.