Effects of drugs on the initiation and maintenance of status epilepticus induced by administration of pilocarpine to lithium-pretreated rats

The ability of various drugs to prevent the onset of status epilepticus induced by administration of the muscarinic agonist, pilocarpine, to lithium-pretreated rats was determined. Motor limbic seizures and status epilepticus occurred in 100% of rats administered pilocarpine (30 mg/kg, s.c.) 20 h after pretreatment with lithium (3 meq/kg, i.p.). The latency to spike activity and to status epilepticus was 20 +/- 1 min and 24 +/- 1 min, respectively. Atropine, diazepam, phenytoin, carbamazepine, phenobarbital, paraldehyde, and L-phenylisopropyladenosine (L-PIA) prevented all phases of seizure activity induced by lithium/pilocarpine treatment. The initiation of status epilepticus was significantly prolonged by pretreatment with sodium valproate. These findings indicate that the seizures induced by administration of lithium and pilocarpine accurately model generalized tonic-clonic epilepsy. The anticonvulsant activity of L-PIA was prevented by prior treatment with the adenosine antagonist, theophylline. The latency to spike and seizure activity was decreased by theophylline, indicating that endogenous adenosine may have a tonic inhibitory influence on cholinergic neurons. Atropine, diazepam, phenobarbital, phenytoin, sodium valproate, L-PIA, and carbamazepine did not interrupt seizure activity when administered 60 min after pilocarpine (approximately 35 min after initiation of status epilepticus). When rats were administered paraldehyde at this time, status epilepticus was rapidly terminated and all rats survived. Thus, status epilepticus induced by lithium and pilocarpine provides a seizure model that is not responsive to conventional anticonvulsants.     

Status epilepticus facilitated by pilocarpine in amygdala-kindled rats

A novel model of status epilepticus based on the kindling model of epilepsy is described. The model involves the administration of a small dose of pilocarpine (20 mg/kg) to rats that have been previously kindled. Stimulation of these pretreated rats produces seizures which continue uninterrupted for approximately 4 h before spontaneous termination. The electroencephalographic discharge pattern showed characteristic changes in polarity and amplitude throughout the duration of status epilepticus. Behaviorally, the animals showed motor seizures which varied between stages I through IV, with evidence of extensive bilateral hemispheric involvement through much of the seizure episode. Animals that had been partially kindled to stage II seizures did not develop status epilepticus after stimulation when pretreated with pilocarpine, indicating that prior kindling is integral to the development of status epilepticus in this model. Administration of scopolamine was ineffective in terminating the condition when it had begun, suggesting that cholinergic stimulation is necessary for the initiation, but not the maintenance, of status epilepticus. This model holds promise for the study of status epilepticus because the condition develops in a seizure-prone (kindled) rat, and the seizures are self-sustaining, without the presence of exogenous chemicals or neurotoxins.     

Repeated low-dose treatment of rats with pilocarpine: low mortality but high proportion of rats developing epilepsy.

Systemic administration of pilocarpine in rats can result in a chronic behavioral state that is similar to human temporal lobe epilepsy. The pilocarpine model of epilepsy is widely used for studying the factors that contribute to the development of epilepsy as a consequence of status epilepticus (SE). For this purpose, pilocarpine is either administered alone at a high systemic dose or in combination with lithium, which markedly potentiates the convulsant effect of pilocarpine. Both experimental protocols, however, are associated with high mortality rates. In the present study, we evaluated whether mortality rate in rats can be decreased by repeated administration of low doses of pilocarpine. The time the rats spent in SE was limited by diazepam. Preliminary experiments in lithium-free rats indicated that repeated low-dose administration of pilocarpine is too time-consuming to produce SE compared to single high-dose administration. All subsequent experiments were performed in lithium-pretreated rats. Single-dose injection of 30 mg/kg pilocarpine produced SE in approximately 70% of the animals, but 45% of the rats died although SE was interrupted by diazepam after 90 min. Repeated i.p. administration of 10 mg/kg pilocarpine at 30-min intervals resulted in SE after 2-4 injections; the mean dose of pilocarpine needed to induce SE was 26 mg/kg. When SE was interrupted after 90 min, mortality rate was below 10%, which was significantly lower compared to the protocol with one single administration of 30 mg/kg pilocarpine. In contrast to mortality rate, the development of spontaneous recurrent seizures did not differ between experimental protocols. Almost all rats which had experienced a SE of at least 60 min developed chronic epilepsy. Average latency to the first spontaneous seizure was approximately 40 days. The frequency and severity of spontaneous seizures was not significantly different between protocols, although animal groups with repeated low-dose treatment tended to have higher frequencies of spontaneous seizures compared to single-dose administration. The present study demonstrates that systemic treatment of lithium-pretreated rats with several low doses of pilocarpine efficiently produces SE and chronic epilepsy with much lower mortality rates than single-dose pilocarpine.     

The course of untreated seizures in the pilocarpine model of epilepsy

The course of untreated epilepsy is not well established. This study uses a model of chronic limbic epilepsy (pilocarpine model of epilepsy) to determine the pattern of occurrence of seizures in untreated animals. Following pilocarpine administration, 21 rats were monitored continuously with a video system for 135 days after the first spontaneous seizure. Animals showed a great variability in seizure numbers and were divided in two subgroups presenting either a low frequency of seizures (n = 9 animals presenting ten or less seizures in the first 15 days of observation) or a high frequency of seizures (n = 12 animals presenting more than ten seizures during this period). Animals with low number of seizures during the first 15 days of observation showed a significant increase in seizure frequency in the following period of analysis (until 105 days). On the other hand, those with initial high number of seizures showed significant changes in seizure frequency only in the first 2 months. The duration of each spontaneous seizure did not change significantly over time. These findings show that in untreated epilepsy there is a maturation process in the early stages and this accelerating process can be of predictive value for the treatment of epilepsy.     

Mapping a mouse limbic seizure susceptibility locus on chromosome 10

Purpose: Mapping seizure susceptibility loci in mice provides a framework for identifying potentially novel candidate genes for human epilepsy. Using C57BL/6J × A/J chromosome substitution strains (CSS), we previously identified a locus on mouse chromosome 10 (Ch10) conferring susceptibility to pilocarpine, a muscarinic cholinergic agonist that models human temporal lobe epilepsy by inducing initial limbic seizures and status epilepticus (status), followed by hippocampal cell loss and delayed‐onset chronic spontaneous limbic seizures. Herein we report further genetic mapping of pilocarpine quantitative trait loci (QTLs) on Ch10. Methods: Seventy‐nine Ch10 F₂ mice were used to map QTLs for duration of partial status epilepticus and the highest stage reached in response to pilocarpine. Based on those results we created interval‐specific congenic lines to confirm and extend the results, using sequential rounds of breeding selectively by genotype to isolate segments of A/J Ch10 genome on a B6 background. Key Findings: Analysis of Ch10 F₂ genotypes and seizure susceptibility phenotypes identified significant, overlapping QTLs for duration of partial status and severity of pilocarpine‐induced seizures on distal Ch10. Interval‐specific Ch10 congenics containing the susceptibility locus on distal Ch10 also demonstrated susceptibility to pilocarpine‐induced seizures, confirming results from the F₂ mapping population and strongly supporting the presence of a QTL between rs13480781 (117.6 Mb) and rs13480832 (127.7 Mb). Significance: QTL mapping can identify loci that make a quantitative contribution to a trait, and eventually identify the causative DNA‐sequence polymorphisms. We have mapped a locus on mouse Ch10 for pilocarpine‐induced limbic seizures. Novel candidate genes identified in mice can be investigated in functional studies and tested for their role in human epilepsy.     

New insights from the use of pilocarpine and kainate models

Local or systemic administration of pilocarpine and kainate in rodents leads to a pattern of repetitive limbic seizures and status epilepticus, which can last for several hours. A latent period follows status epilepticus and precedes a chronic phase, which is characterized by the occurrence of spontaneous limbic seizures. These distinct features, in a single animal preparation, of an acute damage induced by status epilepticus, a silent interval between injury and the onset of spontaneous seizures, and a chronic epileptic state have allowed antiepileptic drug (AED) studies with different purposes, (a) in the acute phase, identification of compounds with efficacy against refractory status epilepticus and/or neuroprotection against damage induced by sustained seizures; (b) in the latent period, identification of agents with a potential for preventing epileptogenesis and/or against seizure-induced long-term behavioral deficits and (c) in the chronic phase, testing drugs effective against partial and secondarily generalized seizures. Studies on pilocarpine and kainate models have pointed out that some AEDs or other compounds exert an antiepileptogenic effect. The analogy of the latent phase of pilocarpine and kainate models with the acquisition of amygdala kindling should encourage testing of drugs that have proved to suppress the evolution of amygdala kindling. Drug testing in the chronic phase should not address only the suppression of secondarily generalized motor seizures. Most of current tools used to quantify spontaneous seizure events need to be coupled to electrophysiology and more sophisticated systems for recording and analyzing behavior.     

癫痫持续状态大鼠模型的GABA_A受体亚单位α1的mRNA表达及[~3H]Flunitrazepam受体配体结合的变化

目的 :本实验研究大鼠癫痫持续状态动物模型的海马等部位 GABAA受体α1亚单位基因表达和受体一配体结合的变化。方法 :成年雄性大鼠经腹腔内注射 32 0～ 34 0± 5 .87毫克 /公斤毛果芸香碱 (Pilocarpine)以制成癫痫持续状态动物模型 ,能在癫痫持续状态 (定义为在皮质脑电图上显示痫性放电的至少 40分钟的持续性痫性发作 )下存活的大鼠在 1小时和 2小时后处于死 ,分别研究 GABA受体基因表达和放射结合位点 ,用原位杂交方法来测定脑部 m RNA水平 ,用 [3H]flunirazepam标记 GABAA 受体 benzodiazepam结合位点。结果 :动物痫性发作 2小时后海马的 CA1和CA3区域 GABAA受体 α1亚单位 m RNA显著下降 ,但是齿状回的 α1m RNA没有变化。 [3H]flunirazepam标记受体 -配体放射结合在持续 2小时持续痫性发作后可见海马的 CA1及 CA3和齿状回中均见下降 ,1小时的持续痫性发作尚未引起海马区域的任何α1m RNA或 [3H]flunirazepam受体 -配体放射结合的任何改变 ,并用结晶染色 1及 2小时后的大脑海马部位。结论 :本研究结果提示大鼠的癫痫持续状态可诱发海马区 GABAA 受体 α1基因表达的改变和 [3H]flinirazepam受体 -配体结合的下降 ,上述改变可能使大脑更容易形成慢性癫痫病灶     

Prolonged febrile seizures in the immature rat model enhance hippocampal excitability long term

Febrile seizures (FSs) constitute the most prevalent seizure type during childhood. Whether prolonged FSs alter limbic excitability, leading to spontaneous seizures (temporal lobe epilepsy) during adulthood, has been controversial. Recent data indicate that, in the immature rat model, prolonged FSs induce transient structural changes of some hippocampal pyramidal neurons and long-term functional changes of hippocampal circuitry. However, whether these neuroanatomical and electrophysiological changes promote hippocampal excitability and lead to epilepsy has remained unknown. By using in vivo and in vitro approaches, we determined that prolonged hyperthermia-induced seizures in immature rats caused long-term enhanced susceptibility to limbic convulsants that lasted to adulthood. Thus, extensive hippocampal electroencephalographic and behavioral monitoring failed to demonstrate spontaneous seizures in adult rats that had experienced hyperthermic seizures during infancy. However, 100% of animals developed hippocampal seizures after systemic administration of a low dose of kainate, and most progressed to status epilepticus. Conversely, a minority of normothermic and hyperthermic controls had (brief) seizures, none developing status epilepticus. In vitro, spontaneous epileptiform discharges were not observed in hippocampal-entorhinal cortex slices derived from either control or experimental groups. However, Schaeffer collateral stimulation induced prolonged, self-sustaining, status epilepticus-like discharges exclusively in slices from experimental rats. These data indicate that hyperthermic seizures in the immature rat model of FSs do not cause spontaneous limbic seizures during adulthood. However, they reduce thresholds to chemical convulsants in vivo and electrical stimulation in vitro, indicating persistent enhancement of limbic excitability that may facilitate the development of epilepsy.     

Malnutrition in infancy as a susceptibility factor for temporal lobe epilepsy in adulthood induced by the pilocarpine experimental model

Malnutrition during the earliest stages of life may result in innumerable brain problems. Moreover, this condition could increase the chances of developing neurological diseases, such as epilepsy. We analyzed the effects of early-life malnutrition on susceptibility to epileptic seizures induced by the pilocarpine model of epilepsy. Wistar rat pups were kept on a starvation regimen from day 1 to day 21 after birth. At day 60, 16 animals (8 = well-nourished; 8 = malnourished) were exposed to the pilocarpine experimental model of epilepsy. Age-matched well-nourished (n = 8) and malnourished (n = 8) rats were used as controls. Animals were video-monitored over 9 weeks. The following behavioral parameters were evaluated: first seizure threshold (acute period of the pilocarpine model); status epilepticus (SE) latency; first spontaneous seizure latency (silent period), and spontaneous seizure frequency during the chronic phase. The cell and mossy fiber sprouting (MFS) density were evaluated in the hippocampal formation. Our results showed that the malnourished animals required a lower pilocarpine dose in order to develop SE (200 mg/kg), lower latency to reach SE, less time for the first spontaneous seizure and higher seizure frequency, when compared to well-nourished pilocarpine rats. Histopathological findings revealed a significant cell density reduction in the CA1 region and intense MFS among the malnourished animals. Our data indicate that early malnutrition greatly influences susceptibility to seizures and behavioral manifestations in adult life. These findings suggest that malnutrition in infancy reduces the threshold for epilepsy and promotes alterations in the brain that persist into adult life.     

Paraventricular nucleus serotonin mediates neurally stimulated adrenocortical secretion

The purpose of this study was to further elucidate the role of serotonin (5-HT) in adrenocortical regulation. The effects of stimulating the frontal cortex and extrahypothalamic limbic structures, on plasma corticosterone (CS) responses, were studied in rats with vehicle or 5,7-dihydroxytryptamine (5,7-DHT) injection into the midbrain raphe nuclei. In another group of rats the neurotoxin was injected locally into the paraventricular nucleus (PVN) in view of its importance in adrenocortical regulation, and the effects of photic and dorsal hippocampal stimulation on plasma CS were studied. 5,7-DHT caused a significant depletion of hypothalamic 5-HT and blocked the rise in plasma CS following the stimulation of the above neural modalities. These studies suggest that the PVN 5-HT mediates the adrenocortical responses following afferent neural stimuli.     

Drug resistance and hippocampal damage after delayed treatment of pilocarpine-induced epilepsy in the rat

Temporal lobe epilepsy (TLE) is the most common and pharmacoresistant form of epilepsy. Problems that cause pharmacoresistance may include delayed therapy due to late consultation, especially in developing countries. Our study aimed at unraveling consequences of delayed drug treatment using a rat model of TLE. Following pilocarpine-induced status epilepticus interrupted after 4h, rats were continuously videorecorded for onset and recurrence of spontaneous convulsive seizures. The animals were then treated for 50 days with carbamazepine (CBZ; first-line drug in TLE and effective also in rats), starting at seizure onset (27.22+/-3.38 days after status epilepticus) or 50 days later, and compared with epileptic untreated rats and non-epileptic CBZ-treated ones. Convulsive seizure frequency and duration, and hippocampal cell changes were evaluated. In particular, parvalbumin-containing hippocampal interneurons, astrocytes and microglia were characterized with immunohistochemistry and quantitative analyses. Prompt administration of CBZ suppressed seizures; delayed treatment only decreased frequency of convulsive seizures, which were also relatively prolonged. In hippocampal regions, histopathological damage, parvalbumin immunoreactivity loss, and glial activation were very marked after delayed treatment, and were reduced only slightly compared to untreated epilepsy, but enhanced compared to early treatment. The data on high frequency and duration of convulsive seizures in late-therapy rats indicate that delayed CBZ administration caused a high degree of drug resistance. This condition was subserved by severe damage in the hippocampus, presumably consequent to long-term seizure recurrence. Overall the data indicate that the paradigm of delayed treatment of limbic epilepsy could provide a model of drug-refractory TLE with hippocampal sclerosis.     

Chronic bilateral stimulation of the anterior thalamus of kainate-treated rats increases seizure frequency

PURPOSE: Electrical stimulation of the anterior nucleus of the thalamus (ANT) is receiving increased attention as a novel means of controlling intractable epilepsy, and has entered human clinical trial. Animal data supporting the anticonvulsant benefit of ANT stimulation, however, has been obtained from acute chemoconvulsant models of epilepsy rather than models of chronic epilepsy with spontaneous seizures. It is unknown whether ANT stimulation is effective in models of chronic epilepsy. METHODS: Bilateral ANT stimulation was evaluated in rats with chronic epilepsy following acute status epilepticus (SE) produced by systemic kainic acid (KA) administration. The evolution of epilepsy following KA SE and the effects of ANT stimulation were monitored by continuous video-EEG. RESULTS: Following KA SE, most rats have 2-8 seizures per day, and the average seizure rate increases over time, doubling over the course of 14 weeks. Behavioral seizure severity, after the initial development of epilepsy, remains stable. Seizure frequency during ANT stimulation was 2.5 times the baseline seizure frequency. In some cases stimulation triggered seizures were observed. The effects of stimulation were specific to the ANT. Stimulation applied to electrodes placed outside the ANT did not significantly worsen seizure frequency. CONCLUSIONS: ANT stimulation exacerbated seizure frequency in rats with chronic epilepsy following kainate status epilepticus.     

Effects of pinealectomy and the treatment with melatonin on the temporal lobe epilepsy in rats

The aim of the present work was to analyze the effects of pinealectomy in the development of the epilepsy model induced by pilocarpine in adult male rats. Group I: Wistar male adult rats were submitted to pinealectomy, and 7 days after surgery, these animals received pilocarpine (350 mg/kg, i.p.) to induce three distinct behavioral phases: status epilepticus, seizure-free, and chronic phases. This late, as well as all control groups were continuously video-recorded for 60 days, to study behavior parameters. These animals were killed and the brain sections were processed for Nissl and neo-Timm. Group II: Another group, also submitted to pinealectomy, received several injections of melatonin (2.5 mg/kg): 20 min before, concomitantly with pilocarpine, 30 min, 1 h, and 2 h after pilocarpine administration. Some animals from group I and all from group II were sacrificed 48 h following status epilepticus onset to perform TUNEL assay. The latency for status epilepticus onset, status epilepticus length as well as mortality rate during status epilepticus were similar for pinealectomized and control groups. On the other hand, pinealectomized rats presented minor duration of the silent period, a higher number of spontaneous seizures during the chronic phase, increased number of TUNEL-positive cells (acute phase), increased neuronal loss, and marked supragranullar mossy fibers sprouting (chronic phase) in the hippocampal formation, when compared with control groups. Our data show that the pinealectomy facilitates the epileptogenic process that follows the long-lasting status epilepticus. This facilitation can be partially reverted by the simultaneous administration of melatonin.     

癫痫持续状态大鼠模型的GABAA受体亚单位α1的mRNA表达及[^3H]Flunitrazepam受体配体结合的变化

目的本实验研究大鼠癫痫持续状态动物模型的海马等部位GABAA受体α1亚单位基因表达和受体一配体结合的变化。方法成年雄性大鼠经腹腔内注射320～340±5.87毫克/公斤毛果芸香碱(Pilocarpine)以制成癫痫持续状态动物模型，能在癫痫持续状态(定义为在皮质脑电图上显示痫性放电的至少40分钟的持续性痫性发作)下存活的大鼠在1小时和2小时后处于死，分别研究GABA受体基因表达和放射结合位点，用原位杂交方法来测定脑部mRNA水平，用［3H］flunirazepam标记GABAA受体benzodiazepam结合位点。结果动物痫性发作2小时后海马的CA1和CA3区域GABAA受体α1亚单位mRNA显著下降，但是齿状回的α1mRNA没有变化。［3H］flunirazepam标记受体-配体放射结合在持续2小时持续痫性发作后可见海马的CA1及CA3和齿状回中均见下降，1小时的持续痫性发作尚未引起海马区域的任何α1mRNA或［3H］flunirazepam受体-配体放射结合的任何改变，并用结晶染色1及2小时后的大脑海马部位。结论本研究结果提示大鼠的癫痫持续状态可诱发海马区GABAA受体α1基因表达的改变和［3H］flinirazepam受体-配体结合的下降，上述改变可能使大脑更容易形成慢性癫痫病灶。     

Superoxide dismutase, glutathione peroxidase activities and the hydroperoxide concentration are modified in the hippocampus of epileptic rats.

The relationship between free radical and scavenger enzymes has been found in the epileptic phenomena and reactive oxygen species have been implicated in seizure-induced neurodegeneration. Using the epilepsy model obtained by systemic administration of pilocarpine (PILO) in rats, we investigated the superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities as well as the hydroperoxide (HPx) concentration in the hippocampus of rats during status epilepticus (SE), silent and chronic periods. The enzyme activities as well as the HPx concentration were measured using spectrophotometric methods and the results compared to values obtained from saline-treated animals. The SOD activity decreased after long-lasting SE period and during the chronic phase. In addition, HPx levels increased in same periods whereas the GPx activity increased only in the hippocampus of animals submitted to 1 h of SE. Animals presenting partial seizures, those submitted to 5 h of SE and animals from the silent period (seizure free) showed normal levels of SOD, GPx and HPx. These results show a direct evidence of lipid peroxidation during seizure activity that could be responsible for neuronal damage in the hippocampus of rats, during the establishment of PILO model of epilepsy.     

Spontaneous Recurrent Seizures in Rats: Amino Acid and Monoamine Determination in the Hippocampus

Summary: Rats subjected to structural brain damage induced by sustained convulsions triggered by systemic administration of pilocarpine (PILO) are a useful model for investigation of the mechanisms essential for seizure generation and spread in rodents. After PILO administration, three distinct phases are observed: (a) an acute period of 1–2 days' duration corresponding to a pattern of repetitive limbic seizures and status epilepticus; (b) a seizurefree (silent) period characterized by a progressive return to normal EEG and behavior of 4–44 days' duration; and (c) a period of spontaneous recurrent seizures (SRS) starting 5-45 days after PILO administration and lasting throughout the animal's life. PILO (320–350 mgikg intraperitoneally, i.p.) was administered to rats, and the content of hippocampal monoamines and amino acids was measured in the acute, silent, and SRS periods by liquid chromatography. Norepinephrine (NE) level was decreased during all periods whereas dopamine (DA) content was increased. Serotonin (5-hydroxytryptamine, 5-HT) was increased only in the acute period. Utilization rate measurement of monoamines showed increased NE consumption and decreased DA consumption during all phases. 5-HT utilization rate was increased only in the acute period. Amino acid content showed a decrease in aspartate (ASP) and glutamate (GLU) concentrations associated with increased y-aminobutyric acid (GABA) level during the acute period. The silent phase was characterized by a decrease in glycine (GLY) and GABA levels and an increase in GLU concentration. The SRS period showed an increase in all amino acid concentrations. These findings show important neurochemical changes in the course of establishment of an epileptic focus after brain damage induced by status epilepticus triggered by pilocarpine.     

Functional Anatomy of Spontaneous Seizures in a Rat Model of Limbic Epilepsy

Summary: Purpose: The substrate of seizure initiation in partial epilepsy is not well understood. Although many studies imply a focus that is will localized to a single structure, some information suggests that seizures arise regionally with simultaneous or near-simultaneous onset at several separate sites. To determine the physiological pattern of seizure onset, recordings were made of spontaneous seizures from multiple limbic structures in a rat model of limbic epilepsy. Methods: Seventeen rats with chronic spontaneous limbic seizures that occurred after an episode of electrical hippocampal stimulation-induced status epilepticus (SE) underwent chronic recording of seizures with a minimum of four electrodes placed bilaterally in the midventral hippocampus and either the amygdala or piriform cortex. As many as 15 of the first recorded spontaneous seizures (mean of 12 seizures, range 2–15) was evaluated for each animal with regard to pattern of seizure onset (focal hippocampal, focal nonhippocampal, or diffuse). The results were evaluated for number of seizure patterns displayed by each animal and the potential change in the pattern of onset with successive seizures (from the first to the fifteenth). Results: In all, 210 seizures were evaluated, beginning 1 week after SE and with monitoring lasting ≥18 weeks (mean recording duration 6 weeks) to record a maximum of 15 seizures for each animal. Overall, 54% of the seizures were of diffuse onset, 25% were of nonhippocampal onset, and 21% were of hippocampal onset. Eight of the 17 (47%) animals had seizures with all three patterns of onset. Evaluation of the evolution of the pattern of onset from the first to the fifteenth seizure recorded for the animals showed a clear tendency for the later seizures to have diffuse onsets (regression analysis r = 0.737, p < 0.002). Qualitative histologic analysis demonstrated neuronal loss in the recorded regions. Conclusions: These findings indicate that the epileptogenic zone is broad in this model of limbic “focal” epilepsy and suggest that the substrate for seizure generation is distributed over several structures.     

Accelerated cognitive decline in a rodent model for temporal lobe epilepsy

ObjectiveCognitive impairment is frequently observed in patients with temporal lobe epilepsy. It is hypothesized that cumulative seizure exposure causes accelerated cognitive decline in patients with epilepsy. We investigated the influence of seizure frequency on cognitive decline in a rodent model for temporal lobe epilepsy.MethodsNeurobehavioral assessment was performed before and after surgery, after the induction of self-sustaining limbic status epilepticus (SSLSE), and in the chronic phase in which rats experienced recurrent seizures. Furthermore, we assessed potential confounders of memory performance.ResultsRats showed a deficit in spatial working memory after the induction of the SSLSE, which endured in the chronic phase. A progressive decline in recognition memory developed in SSLSE rats. Confounding factors were absent. Seizure frequency and also the severity of the status epilepticus were not correlated with the severity of cognitive deficits.SignificanceThe effect of the seizure frequency on cognitive comorbidity in epilepsy has long been debated, possibly because of confounders such as antiepileptic medication and the heterogeneity of epileptic etiologies. In an animal model of temporal lobe epilepsy, we showed that a decrease in spatial working memory does not relate to the seizure frequency. This suggests for other mechanisms are responsible for memory decline and potentially a common pathophysiology of cognitive deterioration and the occurrence and development of epileptic seizures. Identifying this common denominator will allow development of more targeted interventions treating cognitive decline in patients with epilepsy. The treatment of interictal symptoms will increase the quality of life of many patients with epilepsy.     

The pilocarpine model of epilepsy

The systemic administration of a potent muscarinic agonist pilocarpine in rats promotes sequential behavioural and electrographic changes that can be divided in three distinct periods: (a) an acute period that built up progressively into a limbic status epilepticus and that lasts 24h, (b) a silent period with a progressive normalization of EEG and behaviour which varies from 4 to 44 days, and (c) a chronic period with spontaneous recurrent seizures (SRSs). The main features of the SRSs observed during the long-term period resemble those of human complex partial seizures and recurs 2–3 times per week per animal. Therefore, this novel and unique experimental approach may serve as a model of epilepsy mimicking the human condition.This research was supported by FAPESP, CNPq and FINEP from Brazil.     

Cyclicity of spontaneous recurrent seizures in pilocarpine model of temporal lobe epilepsy in rat

Pilocarpine administration to rats results in status epilepticus (SE) and after a latency period to the occurrence of spontaneous seizures. The model is commonly used to investigate mechanisms of epileptogenesis as well as the antiepileptic effects of novel compounds. Surprisingly, there have been no video-EEG studies determining the duration of latency period from SE to the appearance of the first spontaneous seizures or the type and frequency of spontaneous seizures at early phase of pilocarpine-induced epilepsy even though such information is critical for design of such studies. To address these questions, we induced SE with pilocarpine in 29 adult male Wistar rats with cortical electrodes. Rats were continuously video-EEG monitored during SE and up to 23 days thereafter. The first spontaneous seizures occurred 7.2+/-3.6 days after SE. During the follow-up, the mean daily seizure frequency was 2.6+/-1.9, the mean seizure duration 47+/-7 s, and the mean behavioral seizure score 3.2+/-0.9. Typically first seizures were partial (score 1-2). Interestingly, spontaneous seizures occurred in clusters with cyclicity, peaking every 5 to 8 days. These data show that in the pilocarpine model of temporal lobe epilepsy the latency period is short. Because many of the early seizures are partial and the seizures occur in clusters, the true phenotype of epilepsy triggered by pilocarpine-induced SE may be difficult to characterize without continuous long-term video-EEG monitoring. Finally, our data suggest that the model can be used for studies aiming at identifying the mechanisms of seizure clustering.