RESUMEN
AIMS: Neonatal seizures are severe pathologies which may result in long-term neurological consequences. High plasma concentrations of homocysteine - hyperhomocysteinemia (hHCy) - are associated with epilepsy. In the present study, we evaluated susceptibility to seizure of neonatal rats with prenatal hHCy. MAIN METHODS: Prenatal hHCy was induced by feeding females with a high-methionine diet. Experiments were performed on pups during the first three postnatal weeks. Flurothyl-induced epileptic behavior was assessed according to Racine's scale. Epileptiform activity in the hippocampus was recorded using electrophysiological methods. The balance of excitation/inhibition, functional GABAergic inhibition and GABA reversal potential in hippocampal neurons were analyzed. KEY FINDINGS: Rats with hHCy developed more severe stages of behavioral patterns during flurothyl-induced epilepsy with shorter latency. Electrophysiological recordings demonstrated higher background neuronal activity in rats with hHCy. Seizure-like events triggered by flurothyl (in vivo) or 4-aminopyridine (in vitro) showed shorter latency, higher power and amplitude. An increased glutamate/GABA synaptic ratio was shown in the pyramidal neurons of rats with hHCy and more slices demonstrated excitation by isoguvacine, a selective GABA(A) receptor agonist, during the first and second postnatal weeks. The GABA driving force and the reversal potential of GABA(A) currents were more positive during the second postnatal week for hHCy rats. SIGNIFICANCE: The higher susceptibility to seizures in rats with prenatal hHCy due to a shift in the balance of excitation/inhibition toward excitation may underlie the clinical evidence about the association of hHCy with an increased risk of epilepsy.
Asunto(s)
Epilepsia , Hiperhomocisteinemia , Embarazo , Femenino , Ratas , Animales , Animales Recién Nacidos , Flurotilo/farmacología , Hiperhomocisteinemia/complicaciones , Ácido gamma-Aminobutírico/farmacología , Convulsiones/inducido químicamente , Convulsiones/patología , HipocampoRESUMEN
Epilepsy is one of the most common neurological disorders, with individuals having an increased susceptibility of seizures in the first few years of life, making children at risk of developing a multitude of cognitive and behavioral comorbidities throughout development. The present study examined the role of PI3K/Akt/mTOR pathway activity and neuroinflammatory signaling in the development of autistic-like behavior following seizures in the neonatal period. Male and female C57BL/6J mice were administered 3 flurothyl seizures on postnatal (PD) 10, followed by administration of minocycline, the mTOR inhibitor rapamycin, or a combined treatment of both therapeutics. On PD12, isolation-induced ultrasonic vocalizations (USVs) of mice were examined to determine the impact of seizures and treatment on communicative behaviors, a component of the autistic-like phenotype. Seizures on PD10 increased the quantity of USVs in female mice and reduced the amount of complex call types emitted in males compared to controls. Inhibition of mTOR with rapamycin significantly reduced the quantity and duration of USVs in both sexes. Changes in USVs were associated with increases in mTOR and astrocyte levels in male mice, however, three PD10 seizures did not result in enhanced proinflammatory cytokine expression in either sex. Beyond inhibition of mTOR activity by rapamycin, both therapeutics did not demonstrate beneficial effects. These findings emphasize the importance of differences that may exist across preclinical seizure models, as three flurothyl seizures did not induce as drastic of changes in mTOR activity or inflammation as observed in other rodent models.
Asunto(s)
Epilepsia , Factores Inmunológicos/farmacología , Inhibidores mTOR/farmacología , Minociclina/farmacología , Convulsiones , Sirolimus/farmacología , Vocalización Animal/efectos de los fármacos , Animales , Convulsivantes/farmacología , Modelos Animales de Enfermedad , Epilepsia/inducido químicamente , Epilepsia/inmunología , Epilepsia/metabolismo , Epilepsia/fisiopatología , Femenino , Flurotilo/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Convulsiones/inducido químicamente , Convulsiones/inmunología , Convulsiones/metabolismo , Convulsiones/fisiopatología , Factores SexualesRESUMEN
Memory deficits significantly decrease an individual's quality of life and are a pervasive comorbidity of epilepsy. Despite the various distinct processes of memory, the majority of epilepsy research has focused on seizures during the encoding phase of memory, therefore the effects of a seizure on other memory processes is relatively unknown. In the present study, we investigated how a single seizure affects memory reactivation in C57BL/6J adult mice using an associative conditioning paradigm. Initially, mice were trained to associate a tone (conditioned stimulus), with the presence of a shock (unconditioned stimulus). Flurothyl was then administered 1 h before, 1 h after, or 6 h before a memory reactivation trial. The learned association was then assessed by presenting a conditioned stimulus in a new context 24 h or 1 wk after memory reactivation. We found that mice receiving a seizure 1 h prior to reactivation exhibited a deficit in memory 24 h later but not 1 wk later. When mice were administered a seizure 6 h before or 1 h after reactivation, there were no differences in memory between seizure and control animals. Altogether, our study indicates that an acute seizure during memory reactivation leads to a temporary deficit in associative memory in adult mice. These findings suggest that the cognitive impact of a seizure may depend on the timing of the seizure relative to the memory process that is active.
Asunto(s)
Aprendizaje por Asociación/fisiología , Conducta Animal/fisiología , Condicionamiento Clásico/fisiología , Trastornos de la Memoria/fisiopatología , Convulsiones/fisiopatología , Animales , Convulsivantes/farmacología , Modelos Animales de Enfermedad , Miedo/fisiología , Femenino , Flurotilo/farmacología , Masculino , Trastornos de la Memoria/etiología , Ratones Endogámicos C57BL , Convulsiones/inducido químicamente , Convulsiones/complicaciones , Factores de TiempoRESUMEN
OBJECTIVE: BRD2 is a human gene repeatedly linked to and associated with juvenile myoclonic epilepsy (JME). Here, we define the developmental stage when increased seizure susceptibility first manifests in heterozygous Brd2+/- mice, an animal model of JME. We wanted to determine (1) whether seizure susceptibility correlates with the proven decrease of γ-aminobutyric acidergic (GABAergic) neuron numbers and (2) whether the seizure phenotype can be affected by sex hormones. METHODS: Heterozygous (Brd2+/-) and wild-type (wt) mice of both sexes were tested for flurothyl-induced seizure susceptibility at postnatal day 15 (P15; wt, n = 13; Brd2+/-, n = 20), at P30 (wt, n = 20; Brd2+/-, n = 20), and in adulthood (5-6 months of age; wt, n = 10; Brd2+/-, n = 12). We measured latency to clonic and tonic-clonic seizure onset (flurothyl threshold). We also compared relative density of parvalbumin-positive (PVA+) and GAD67+ GABA neurons in the striatum and primary motor (M1) neocortex of P15 (n = 6-13 mice per subgroup) and P30 (n = 7-10 mice per subgroup) mice. Additional neonatal Brd2+/- mice were injected with testosterone propionate (females) or formestane (males) and challenged with flurothyl at P30. RESULTS: P15 Brd2+/- mice showed no difference in seizure susceptibility compared to P15 wt mice. However, even at this early age, Brd2+/- mice showed fewer PVA+ neurons in the striatum and M1 neocortex. Compared to wt, the striatum in Brd2+/- mice showed an increased proportion of immature PVA+ neurons, with smaller cell bodies and limited dendritic arborization. P30 Brd2+/- mice displayed increased susceptibility to flurothyl-induced clonic seizures compared to wt. Both genotype and sex strongly influenced the density of PVA+ neurons in the striatum. Susceptibility to clonic seizures remained increased in adult Brd2+/- mice, and additionally there was increased susceptibility to tonic-clonic seizures. In P30 females, neonatal testosterone reduced the number of flurothyl-induced clonic seizures. SIGNIFICANCE: A decrease in striatal PVA+ GABAergic neurons developmentally precedes the onset of increased seizure susceptibility and likely contributes to the expression of the syndrome.
Asunto(s)
Flurotilo/farmacología , Epilepsia Mioclónica Juvenil/patología , Neuronas/patología , Parvalbúminas/metabolismo , Convulsiones/inducido químicamente , Androstenodiona/análogos & derivados , Androstenodiona/farmacología , Animales , Modelos Animales de Enfermedad , Femenino , Neuronas GABAérgicas/efectos de los fármacos , Neuronas GABAérgicas/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Epilepsia Mioclónica Juvenil/inducido químicamente , Neuronas/efectos de los fármacos , Convulsiones/patología , Propionato de Testosterona/farmacología , Factores de Transcripción/metabolismoRESUMEN
OBJECTIVE: Recurrent seizures can cause brain damage and affect the cognitive outcome, particularly in developing children. We aimed to determine the effects of recurrent seizures on the expression of gamma-aminobutyric acid A receptor (GABAAR) α1 and γ2 subunit and neurodevelopment in immature rats. The role of the GABAAR agonist clonazepam and antagonist/partial agonist flumazenil in seizure-induced brain injury was also studied. METHODS: Recurrent seizures (RS) were induced by flurothyl inhalation in immature rats. Clonazepam (CZP) and flumazenil (FMZ) were administered to modulate GABAAR subunit expression in different experimental groups. Neurobehavioral changes and GABAAR α1 and γ2 subunit expression were studied. RESULTS: Inhalation of flurothyl for five days triggered RS and caused reflex delay, inability to adapt to new environments in adulthood, and deficits in long-term learning and memory ability in rats. Down-regulation of GABAAR α1 and γ2 subunits occurred after seizure onset and persisted for a long time. CZP treatment decreased the expression of GABAAR α1 and γ2 subunits and delayed neurodevelopment of the immature rats, whereas FMZ did not show any significant effects. CONCLUSIONS: Changes in GABAAR α1 and γ2 subunit expression and neurodevelopment were related to recurrent seizures and administration of CZP. Thus, GABAAR α1 and γ2 subunits likely play a significant role in the development of immature rats with RS and provide a novel target for therapeutic intervention.
Asunto(s)
Lesiones Encefálicas/tratamiento farmacológico , Epilepsia Generalizada/tratamiento farmacológico , Hipocampo/efectos de los fármacos , Receptores de GABA-A/metabolismo , Convulsiones/metabolismo , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Lesiones Encefálicas/metabolismo , Epilepsia Generalizada/metabolismo , Femenino , Flurotilo/farmacología , Hipocampo/metabolismo , Masculino , Ratas Wistar , Convulsiones/tratamiento farmacológico , Ácido gamma-Aminobutírico/metabolismo , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
Rapid antidepressant effects of ketamine become most evident when its psychotomimetic effects subside, but the neurobiological basis of this "lag" remains unclear. Laughing gas (N2O), another NMDA-R (N-methyl-D-aspartate receptor) blocker, has been reported to bring antidepressant effects rapidly upon drug discontinuation. We took advantage of the exceptional pharmacokinetic properties of N2O to investigate EEG (electroencephalogram) alterations and molecular determinants of antidepressant actions during and immediately after NMDA-R blockade. Effects of the drugs on brain activity were investigated in C57BL/6 mice using quantitative EEG recordings. Western blot and qPCR were used for molecular analyses. Learned helplessness (LH) was used to assess antidepressant-like behavior. Immediate-early genes (e.g., bdnf) and phosphorylation of mitogen-activated protein kinase-markers of neuronal excitability-were upregulated during N2O exposure. Notably, phosphorylation of BDNF receptor TrkB and GSK3ß (glycogen synthase kinase 3ß) became regulated only gradually upon N2O discontinuation, during a brain state dominated by slow EEG activity. Subanesthetic ketamine and flurothyl-induced convulsions (reminiscent of electroconvulsive therapy) also evoked slow oscillations when their acute pharmacological effects subsided. The correlation between ongoing slow EEG oscillations and TrkB-GSK3ß signaling was further strengthened utilizing medetomidine, a hypnotic-sedative agent that facilitates slow oscillations directly through the activation of α2-adrenergic autoreceptors. Medetomidine did not, however, facilitate markers of neuronal excitability or produce antidepressant-like behavioral changes in LH. Our results support a hypothesis that transient cortical excitability and the subsequent regulation of TrkB and GSK3ß signaling during homeostatic emergence of slow oscillations are critical components for rapid antidepressant responses.
Asunto(s)
Antidepresivos/farmacología , Corteza Cerebral/metabolismo , Electroencefalografía , Neuronas/metabolismo , Receptor trkB/metabolismo , Transducción de Señal , Anestésicos/farmacología , Animales , Biomarcadores/metabolismo , Corteza Cerebral/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Flurotilo/farmacología , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Homeostasis/efectos de los fármacos , Ketamina/farmacología , Medetomidina/farmacología , Ratones Endogámicos C57BL , Neuronas/efectos de los fármacos , Óxido Nitroso/farmacología , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Using an experimental model of neonatal recurrent seizures we investigated the influence of epileptic seizures in the various forms of synaptic plasticity in neurons of the somatosensory cortex. We found that early seizures do not affect the post-tetanic potentiation of the amplitude of the postsynaptic potentials and the depression of postsynaptic potentials during high-frequency stimulation. However they result in the chronic increase of the long-term potentiation of synaptic transmission. These changes of synaptic plasticity may affect the processing of the sensory information in patients with a history of recurrent seizures during early development.
Asunto(s)
Epilepsia/fisiopatología , Potenciación a Largo Plazo/efectos de los fármacos , Corteza Somatosensorial/fisiopatología , Transmisión Sináptica/efectos de los fármacos , Animales , Animales Recién Nacidos , Convulsivantes/farmacología , Epilepsia/inducido químicamente , Epilepsia/patología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Flurotilo/farmacología , Humanos , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Masculino , Neuronas/efectos de los fármacos , Neuronas/patología , Ratas Sprague-Dawley , Corteza Somatosensorial/efectos de los fármacos , Corteza Somatosensorial/patologíaRESUMEN
Identifying the genetic basis of epilepsy in humans is difficult due to its complexity, thereby underlying the need for preclinical models with specific aspects of seizure susceptibility that are tractable to genetic analyses. In the repeated-flurothyl model, mice are given 8 flurothyl-induced seizures, once per day (the induction phase), followed by a 28-day rest period (incubation phase) and final flurothyl challenge. This paradigm allows for the tracking of multiple phenotypes including: initial generalized seizure threshold, decreases in generalized seizure threshold with repeated flurothyl exposures, and changes in the complexity of seizures over time. Given the responses we previously reported in C57BL/6J mice, we analyzed substrains of the C57BL lineage to determine if any of these phenotypes segregated in these substrains. We found that the generalized seizure thresholds of C57BL/10SNJ and C57BL/10J mice were similar to C57BL/6J mice, whereas C57BL/6NJ and C57BLKS/J mice showed lower generalized seizure thresholds. In addition, C57BL/6J mice had the largest decreases in generalized seizure thresholds over the induction phase, while the other substrains were less pronounced. Notably, we observed only clonic seizures during the induction phase in all substrains, but when rechallenged with flurothyl after a 28-day incubation phase, â¼80% of C57BL/6J and 25% of C57BL/10SNJ and C57BL/10J mice expressed more complex seizures with tonic manifestations with none of the C57BL/6NJ and C57BLKS/J mice having complex seizures with tonic manifestations. These data indicate that while closely related, the C57BL lineage has significant diversity in aspects of epilepsy that are genetically controlled. Such differences further highlight the importance of genetic background in assessing the effects of targeted deletions of genes in preclinical epilepsy models.
Asunto(s)
Modelos Animales de Enfermedad , Epilepsia/genética , Flurotilo/farmacología , Convulsiones/inducido químicamente , Análisis de Varianza , Animales , Cruzamientos Genéticos , Relación Dosis-Respuesta a Droga , Masculino , Ratones , Ratones Endogámicos C57BL , Convulsiones/genética , Especificidad de la EspecieRESUMEN
The Kv2.1 delayed rectifier potassium channel exhibits high-level expression in both principal and inhibitory neurons throughout the central nervous system, including prominent expression in hippocampal neurons. Studies of in vitro preparations suggest that Kv2.1 is a key yet conditional regulator of intrinsic neuronal excitability, mediated by changes in Kv2.1 expression, localization and function via activity-dependent regulation of Kv2.1 phosphorylation. Here we identify neurological and behavioral deficits in mutant (Kv2.1(-/-) ) mice lacking this channel. Kv2.1(-/-) mice have grossly normal characteristics. No impairment in vision or motor coordination was apparent, although Kv2.1(-/-) mice exhibit reduced body weight. The anatomic structure and expression of related Kv channels in the brains of Kv2.1(-/-) mice appear unchanged. Delayed rectifier potassium current is diminished in hippocampal neurons cultured from Kv2.1(-/-) animals. Field recordings from hippocampal slices of Kv2.1(-/-) mice reveal hyperexcitability in response to the convulsant bicuculline, and epileptiform activity in response to stimulation. In Kv2.1(-/-) mice, long-term potentiation at the Schaffer collateral - CA1 synapse is decreased. Kv2.1(-/-) mice are strikingly hyperactive, and exhibit defects in spatial learning, failing to improve performance in a Morris Water Maze task. Kv2.1(-/-) mice are hypersensitive to the effects of the convulsants flurothyl and pilocarpine, consistent with a role for Kv2.1 as a conditional suppressor of neuronal activity. Although not prone to spontaneous seizures, Kv2.1(-/-) mice exhibit accelerated seizure progression. Together, these findings suggest homeostatic suppression of elevated neuronal activity by Kv2.1 plays a central role in regulating neuronal network function.
Asunto(s)
Potenciales de Acción , Eliminación de Gen , Neuronas/fisiología , Fenotipo , Convulsiones/genética , Canales de Potasio Shab/metabolismo , Animales , Convulsivantes/farmacología , Flurotilo/farmacología , Hipocampo/citología , Hipocampo/efectos de los fármacos , Hipocampo/fisiología , Potenciación a Largo Plazo , Aprendizaje por Laberinto , Ratones , Ratones Endogámicos C57BL , Neuronas/metabolismo , Pilocarpina/farmacología , Convulsiones/fisiopatología , Canales de Potasio Shab/genéticaRESUMEN
Febrile seizure (FS) remains the most common childhood neurological emergency. Although many studies have been done, controversy exists as to whether these seizures are associated with a significant risk for cognitive impairment. The aim of our study is to check whether there is a spatial learning and memory deficit in the experimental FS rats using a heated-air FS paradigm and to determine the possible molecular mechanism of cognitive impairment. On days 10 to 12 postpartum, the male rat pups were subjected to one, three, or nine episodes of brief hyperthermia-induced seizures (HS). At adolescence and adulthood, the rats subjected to three, or nine episodes of HS had significant deficits in spatial learning and memory tested by Morris water maze. At adulthood, no apparent hippocampal neuronal loss was found in any HS group, but the seizure threshold to flurothyl was decreased significantly in the rats subjected to nine episodes of HS. In the rats subjected to three, or nine episodes of HS, the Western immunoblotting showed that there was a significant translocation of Ca(2+)-calmodulin stimulated protein kinase II (CaMKII) from the postsynaptic density to the cytosol. In the postsynaptic density the phosphorylation of CaMKIIα Thr(286) was reduced significantly, but the phosphorylation of CaMKIIα Thr(305) was increased significantly. Our study showed early-life brief but recurrent HS caused long-term cognitive impairment and CaMKIIα was involved in carrying forward the signal resulting from HS. The change of the phosphorylative level in Thr(286) and Thr(305) sites of CaMKIIα may underlie the molecular mechanism for the HS related cognitive impairment.
Asunto(s)
Proteínas Portadoras/metabolismo , Hipocampo/fisiopatología , Aprendizaje por Laberinto/fisiología , Memoria/fisiología , Convulsiones Febriles/fisiopatología , Animales , Western Blotting , Proteínas de Unión al Calcio , Convulsivantes/farmacología , Modelos Animales de Enfermedad , Flurotilo/farmacología , Hipocampo/crecimiento & desarrollo , Hipocampo/patología , Calor , Masculino , Neuronas/patología , Neuronas/fisiología , Pruebas Neuropsicológicas , Fosforilación , Estimulación Física , Densidad Postsináptica/fisiología , Ratas , Ratas Sprague-Dawley , Convulsiones/inducido químicamente , Convulsiones/etiología , Convulsiones/fisiopatología , Convulsiones Febriles/patología , Percepción Espacial/fisiologíaRESUMEN
PURPOSE: There is a gap in our knowledge of the factors that modulate the predisposition to seizures following perinatal hypoxia. Herein, we investigate in a mouse model the effects of two distinct factors: developmental stage after the occurrence of the perinatal insult, and the presence of a seizure predisposing mutation. METHODS: Effects of age: P6 (postnatal day 6) mouse pups were subjected to acute hypoxia down to 4% O2 over the course of 45 min. Seizure susceptibilities to flurothyl-induced seizures (single exposures) and to flurothyl kindling were determined at specific subsequent ages. Effects of mutation: Heterozygote mice, with deletion of one copy of the Kcn1a gene, subjected to P6 hypoxia were compared as adults to wild-type mice with respect to susceptibility to a single exposure to flurothyl and to the occurrence of spontaneous seizures as detected by hippocampal electroencephalography (EEG) and video recordings. KEY FINDINGS: Effects of age: As compared to controls, wild-type mice exposed to P6 hypoxia had a shortened seizure latency in response to a single flurothyl exposure at P50, but not at P7 or P28 (p < 0.04). In addition, perinatal hypoxia at P6 enhanced the rate of development of flurothyl kindling performed at P28-38 (p < 0.03), but not at P7-17. Effects of mutation: Kcn1a heterozygous mice subjected to P6 hypoxia exhibited increased susceptibility to flurothyl-induced seizures at P50 as compared to Normoxia heterozygote littermates, and to wild-type Hypoxia and Normoxia mice. In addition, heterozygotes exposed to P6 hypoxia were the only group in which spontaneous seizures were detected during the period of long-term monitoring (p < 0.027 in all comparisons). SIGNIFICANCE: Our data establish a mouse model of mild perinatal hypoxia in which we document the following: (1) the emergence, after a latent period, of increased susceptibility to flurothyl-induced seizures, and to flurothyl induced kindling; and (2) an additive effect of a gene mutation to the seizure predisposing consequences of perinatal hypoxia, thereby demonstrating that a modifier (or susceptibility) gene can exacerbate the long-term consequences of hypoxic injury.
Asunto(s)
Predisposición Genética a la Enfermedad/genética , Hipoxia/complicaciones , Canal de Potasio Kv.1.1/genética , Convulsiones/etiología , Factores de Edad , Animales , Animales Recién Nacidos/genética , Animales Recién Nacidos/fisiología , Modelos Animales de Enfermedad , Electroencefalografía , Flurotilo/farmacología , Heterocigoto , Hipocampo/fisiopatología , Humanos , Excitación Neurológica/efectos de los fármacos , Excitación Neurológica/fisiología , Canal de Potasio Kv.1.1/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Mutación/genética , Convulsiones/inducido químicamente , Convulsiones/genética , Convulsiones/fisiopatologíaRESUMEN
PURPOSE: We evaluated the ability of the ketogenic diet (KD) to improve thresholds to flurothyl-induced seizures in two mouse lines with Scn1a mutations: one that models Dravet syndrome (DS) and another that models genetic (generalized) epilepsy with febrile seizures plus (GEFS+). METHODS: At postnatal day 21, mouse models of DS and GEFS+ were fasted for 12-14 h and then placed on either a 6:1 (fats to proteins and carbohydrates) KD or a standard diet (SD) for 2 weeks. At the end of the 2-week period, we measured thresholds to seizures induced by the chemiconvulsant flurothyl. Body weight, ß-hydroxybutyrate (BHB) levels, and glucose levels were also recorded every 2 days over a 2-week period in separate cohorts of mutant and wild-type mice that were either on the KD or the SD. KEY FINDINGS: Mice on the KD gained less weight and exhibited significantly higher BHB levels compared to mice on the SD. It is notable that thresholds to flurothyl-induced seizures were restored to more normal levels in both mouse lines after 2 weeks on the KD. SIGNIFICANCE: These results indicate that the KD may be an effective treatment for refractory patients with SCN1A mutations. The availability of mouse models of DS and GEFS+ also provides an opportunity to better understand the mechanism of action of the KD, which may facilitate the development of improved treatments.
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Dieta Cetogénica , Epilepsia Generalizada/dietoterapia , Ácido 3-Hidroxibutírico/sangre , Animales , Convulsivantes/farmacología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Epilepsia Generalizada/genética , Canales Epiteliales de Sodio/genética , Flurotilo/farmacología , Ratones , Ratones Noqueados/genética , Ratones Noqueados/fisiología , Convulsiones/inducido químicamente , Convulsiones/prevención & control , Aumento de PesoRESUMEN
Frequent interictal spikes are a common finding in the electroencephalograms of children with epileptic encephalopathies. While it is well recognized that interictal spikes are a biological marker of seizures and can lead to transitory cognitive impairment, whether interictal spikes can result in long-standing adverse effects on learning and memory in children is not known. Here we investigated the consequences of interictal spikes in rat pups without seizures on long-term learning and memory. Rat pups were given a low dose of flurothyl for 4h for 10 days during continuous electroencephalographic monitoring. Rats developed interictal spikes without seizures while age-matched controls under similar testing conditions had few interictal spikes. When rats were tested as adults, there was impairment in reference memory in the probe test of the Morris water maze, reference memory impairment in the four-trial radial-arm water maze and impaired long-term potentiation. Early-life interictal spikes resulted in impaired new cell formation and decreased cell counts in the hippocampus but did not cause an increase in apoptosis. This study, for the first time demonstrates that interictal spikes in rat pups without seizures can result in long-standing spatial cognitive impairment. Our findings suggest that suppressing IIS may be as important as treating seizures during brain development.
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Corteza Cerebral/fisiopatología , Flurotilo/farmacología , Hipocampo/fisiopatología , Aprendizaje por Laberinto/fisiología , Memoria/fisiología , Análisis de Varianza , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Recuento de Células , Corteza Cerebral/efectos de los fármacos , Convulsivantes/farmacología , Electrodos Implantados , Electroencefalografía , Hipocampo/efectos de los fármacos , Etiquetado Corte-Fin in Situ , Potenciación a Largo Plazo/efectos de los fármacos , Potenciación a Largo Plazo/fisiología , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Memoria/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Conducta Espacial/efectos de los fármacos , Conducta Espacial/fisiologíaRESUMEN
Myoclonus is often observed in epilepsy. It is characterized by sudden involuntary shock-like movements of the body (myoclonic jerks, MJs). This study examined whether epileptic myoclonus was under genetic control. Inbred strains of mice were administered eight daily flurothyl exposures, a 28-day rest period, and a final flurothyl retest. For all trials, the latency to the first MJ (threshold) and the number of MJs (MJ#) were recorded. The inbred strains that we examined exhibited significant variability in initial myoclonic response, and myoclonus across the eight flurothyl exposures. C57BL/6J and DBA/2J mice displayed significantly different initial latencies to a MJ, MJ# preceding a generalized seizure (GS), and changes in MJ threshold and MJ# across the eight seizure trials. [C57BL/6J x DBA/2J] F1-hybrid mice showed an initial MJ threshold and decreases in MJ threshold over the eight trials, which were similar to C57BL/6J; however, F1-hybrids had an initial MJ# and trend in MJ# over the eight trials that were similar to DBA/2J. Decreases in MJ threshold and MJ# following multiple seizure trials, observed in C57BL/6J mice, were dependent on the expression of GSs and not on MJ occurrence. Our study is the first to document the potential for genetic heterogeneity of myoclonus in mice; we show that significant alterations in myoclonic behavior occur after GSs. These results indicate that multiple GSs affect MJ thresholds. An understanding of the genetics of myoclonus will be important for determination of the brain areas responsible for myoclonus as well as for identification of candidate genes.
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Flurotilo/farmacología , Ratones Endogámicos/genética , Mioclonía/genética , Análisis de Varianza , Animales , Convulsivantes/farmacología , Cruzamientos Genéticos , Masculino , Ratones , Mioclonía/inducido químicamente , Especificidad de la Especie , Factores de TiempoRESUMEN
Neonatal seizures can result in long-term adverse consequences including alteration of seizure susceptibility and impairment in spatial memory. However, little is known about the effects of neonatal seizures on developmental changes occurring in synaptic transmission during the first postnatal weeks. The purpose of the present study was to examine the effect of neonatal seizures on several aspects of gamma-aminobutyric acid (GABA)ergic and glutamatergic synaptic transmission in the developing rat hippocampus. Flurothyl was used to induce multiple recurrent seizures in rat pups during the first postnatal days. Whole-cell patch-clamp recordings from the hippocampal CA3 pyramidal cell and extracellular recordings from the CA3 pyramidal cell layer were made in slice preparations. In rats that experienced neonatal seizures the amplitude of spontaneous inhibitory postsynaptic currents at P15-17 was decreased by 27% compared with controls, whereas neither frequency nor the kinetic properties were altered. Neonatal seizures did not affect the timing of the developmental switch in the GABAA signaling from excitatory to inhibitory. None of the studied parameters of glutamatergic postsynaptic currents was different between the flurothyl and control groups, including the amplitude and frequency of the spontaneous excitatory postsynaptic currents, the ratio of the amplitudes and frequencies of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA)-mediated spontaneous postsynaptic currents, and the kinetics of AMPA and NMDA mediated postsynaptic currents in the age groups P8-10 and P15-17. We suggest that the selective depression of the amplitude of GABAergic synaptic responses may contribute to the adverse neurological and behavioral consequences that occur following neonatal seizures.
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Animales Recién Nacidos/fisiología , Convulsivantes/farmacología , Flurotilo/farmacología , Convulsiones/fisiopatología , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/fisiología , Animales , Interpretación Estadística de Datos , Potenciales Postsinápticos Excitadores/fisiología , Espacio Extracelular/fisiología , Ácido Glutámico/fisiología , Técnicas In Vitro , Cinética , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , Receptores AMPA/fisiología , Receptores de GABA-A/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Convulsiones/inducido químicamenteRESUMEN
Structures responsible for the onset, propagation, and cessation of generalized seizures are not known. Lesion and microinfusion studies suggest that the substantia nigra pars reticulata (SNR) seizure-controlling network could play a key role. However, the expression of neural activity within the SNR and its targets during discrete pre- and postictal periods has not been investigated. In rats, we used flurothyl to induce generalized seizures over a controlled time period and 2-deoxyglucose autoradiography mapping technique. Changes in neural activity within the SNR were region-specific. The SNRposterior was selectively active during the pre-clonic period and may represent an early gateway to seizure propagation. The SNRanterior and superior colliculus changed their activity during progression to tonic-clonic seizure, suggesting the involvement in coordinated regional activity that results in inhibitory effects on seizures. The postictal suppression state was correlated with changes in the SNR projection targets, specifically the pedunculopontine tegmental nucleus and superior colliculus.
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Epilepsia/fisiopatología , Vías Nerviosas/fisiopatología , Neuronas/fisiología , Sustancia Negra/fisiopatología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Autorradiografía , Mapeo Encefálico , Convulsivantes/farmacología , Desoxiglucosa/metabolismo , Modelos Animales de Enfermedad , Metabolismo Energético/efectos de los fármacos , Metabolismo Energético/fisiología , Epilepsia/inducido químicamente , Flurotilo/farmacología , Masculino , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/metabolismo , Neuronas/efectos de los fármacos , Núcleo Tegmental Pedunculopontino/efectos de los fármacos , Núcleo Tegmental Pedunculopontino/metabolismo , Núcleo Tegmental Pedunculopontino/fisiopatología , Ratas , Ratas Sprague-Dawley , Sustancia Negra/efectos de los fármacos , Sustancia Negra/metabolismo , Colículos Superiores/efectos de los fármacos , Colículos Superiores/metabolismo , Colículos Superiores/fisiopatología , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Núcleos Talámicos Ventrales/efectos de los fármacos , Núcleos Talámicos Ventrales/metabolismo , Núcleos Talámicos Ventrales/fisiopatologíaRESUMEN
PURPOSE: Status epilepticus (SE) commonly occurs in children, whereas ischemic stroke is the most frequent neurologic insult in adults. The purpose of this study was to determine the effect of SE induced in immature (15 days old; PN15) male rats, on susceptibility to subsequent transient focal cerebral ischemia induced in adulthood. METHODS: SE was induced by flurothyl ether (FE) or kainic acid (KA). Rats that did not develop seizures after FE or KA served as controls. Five weeks later, the now-adult rats were subjected to middle cerebral artery occlusion (MCAo) for 1 or 2 h by using the intraluminal filament technique. The extent of the infarct volume was evaluated 24 h later. RESULTS: In rats submitted to 1-h-long FE-SE, the volume of infarction was significantly reduced compared with that in rats exposed to FE without SE. Longer duration of FE-SE was acutely lethal. KA-SE induced prolonged behavioral SE (156 +/- 17.5 min). In these rats, the volume of infarction was significantly larger compared with that in rats that did not show any electrographic seizures after KA administration. Comparison of FE and KA groups revealed that differences in the size of infarction were confined into cortical areas served by the MCA. Neither type of SE induced any obvious histologic changes in these neocortical regions before stroke induction. CONCLUSIONS: Early in life, SE can influence the outcome of a subsequent focal ischemic insult in adulthood. The extent of the infarct is related to the duration and cause of SE. Prolonged SE induced by KA worsens the outcome, whereas FE-SE has a neuroprotective effect.
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Isquemia Encefálica/epidemiología , Estado Epiléptico/inducido químicamente , Factores de Edad , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Conducta Animal/efectos de los fármacos , Conducta Animal/fisiología , Isquemia Encefálica/patología , Infarto Cerebral/epidemiología , Infarto Cerebral/etiología , Infarto Cerebral/patología , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Electroencefalografía , Flurotilo/farmacología , Infarto de la Arteria Cerebral Media/complicaciones , Ácido Kaínico/farmacología , Masculino , Monitoreo Fisiológico , Neocórtex/patología , Fármacos Neuroprotectores/farmacología , Ratas , Ratas Sprague-Dawley , Estado Epiléptico/patología , Accidente Cerebrovascular/epidemiología , Accidente Cerebrovascular/etiologíaRESUMEN
Brain-derived neurotrophic factor (BDNF) appears to be both regulated by and a regulator of epileptogenesis. In the flurothyl (HFE) model of kindling mice exposed to successive flurothyl trials over 8 days express a rapid, long-lasting reduction in generalized seizure threshold and a more slowly evolving change in seizure phenotype in response to subsequent flurothyl exposure. The BDNF genotype of particular mouse strains appears to influence the epileptogenic progression in this model. Thus, we hypothesized that BDNF signaling pathways are altered by flurothyl-induced seizures. Following HFE kindling, fully kindled (eight seizures) adult male C57BI/6J mice had significantly elevated whole brain BDNF levels through at least 28 days after their final seizure. Mice that received only four HFE seizures (not kindled) had elevated BDNF levels, but only at 1 day post-seizure (DPSz), while BDNF levels were not significantly altered in mice receiving just one HFE seizure at any time point studied. Regional expression patterns of BDNF in the hippocampus, hypothalamus, and frontal cortex were also elevated by one DPSz and returned to control values by 14 DPSz in mice that received four HFE seizures. No changes were seen in the cerebellum, striatum, or piriform cortex. In contrast, fully kindled mice had significantly elevated BDNF levels within the hippocampus, hypothalamus, neocortex, and striatum that remained elevated through at least 14 DPSz, while levels were unchanged in the cerebellum and piriform cortex. Regional results were confirmed using anti-BDNF immunohistochemistry (IHC). Despite changes in BDNF levels following HFE kindling, we were unable to demonstrate alterations either in full-length tyrosine kinase receptor B (TrkB) expression (Western blot and IHC) or in truncated TrkB (IHC) expression levels. Together, these data suggest a model of a positive feedback loop involving seizure activity and seizure number and persistently modified BDNF signaling pathways that influences seizure phenotypes within the HFE kindling paradigm. Thus, long-term elevations in BDNF may be responsible in part for epileptogenic processes and the development of human refractory epilepsies.
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Factor Neurotrófico Derivado del Encéfalo/metabolismo , Encéfalo/metabolismo , Epilepsia/metabolismo , Excitación Neurológica/fisiología , Regulación hacia Arriba/fisiología , Animales , Encéfalo/fisiopatología , Modelos Animales de Enfermedad , Epilepsia/inducido químicamente , Epilepsia/fisiopatología , Flurotilo/farmacología , Inmunohistoquímica , Excitación Neurológica/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Fenotipo , Receptor trkB/metabolismo , Transducción de Señal/fisiologíaRESUMEN
UNLABELLED: Several reports suggest that clinically used concentrations of inhaled anesthetics can increase conductance through noninactivating potassium channels and that the resulting hyperpolarization might decrease excitability, thereby leading to the anesthetic state. We speculated that animals deficient in such potassium channels might be resistant to the effects of anesthetics. Thus, in the present study, we measured the minimum alveolar anesthetic concentration (MAC) needed to prevent movement in response to a noxious stimulus in 50% of adult mice lacking functional KCNK5 potassium channel subunits and compared these results with those for heterozygous and wild-type mice. We also measured MAC in weaver mice that had a mutation in the potassium channel Kir3.2 and compared the resulting values with those for wild-type mice. MAC values for desflurane, halothane, and isoflurane for KCNK5-deficient mice and isoflurane MAC values for weaver mice did not differ from MAC values found in control mice. Our results do not support the notion that these potassium channels mediate the capacity of inhaled anesthetics to produce immobility. In addition, we found that the weaver mice did not differ from control mice in their susceptibility to convulsions from the nonimmobilizers flurothyl [di-(2,2,2,-trifluoroethyl)ether] or 2N (1,2-dichlorohexafluorocyclobutane). IMPLICATIONS: Mice harboring mutations in either of two different potassium channels have minimum alveolar anesthetic concentration (MAC) values that do not differ from MAC values found in control mice. Such findings do not support the notion that these potassium channels mediate the capacity of inhaled anesthetics to produce immobility in the face of noxious stimulation.
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Anestésicos por Inhalación/farmacología , Canales de Potasio de Rectificación Interna/genética , Canales de Potasio de Dominio Poro en Tándem , Canales de Potasio/genética , Alveolos Pulmonares/metabolismo , Animales , Células Cultivadas , Convulsivantes/farmacología , ADN/genética , ADN/metabolismo , Electroporación , Flurotilo/farmacología , Vectores Genéticos , Halotano/farmacología , Isoflurano/farmacología , Ratones , Ratones Mutantes Neurológicos , Mutación/genética , Mutación/fisiología , Canales de Potasio/efectos de los fármacos , Canales de Potasio de Rectificación Interna/efectos de los fármacos , Alveolos Pulmonares/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
UNLABELLED: In animals, the conventional inhaled anesthetic, isoflurane, impairs learning fear to context and fear to tone, doing so at concentrations that produce amnesia in humans. Nonimmobilizers are inhaled compounds that do not produce immobility in response to noxious stimulation, nor do they decrease the requirement for conventional inhaled anesthetics. Like isoflurane, the nonimmobilizer 1,2-dichlorohexafluorocyclobutane (2N) impairs learning at concentrations less than those predicted from its lipophilicity to produce anesthesia. The capacity of the nonimmobilizer di-(2,2,2,-trifluoroethyl) ether (flurothyl) to affect learning and memory has not been studied. Both nonimmobilizers can cause convulsions. We hypothesized that if isoflurane, 2N, and flurothyl act by the same mechanism to impair learning and memory, their effects should be additive. We found that isoflurane, 2N, and flurothyl (each, alone) impaired learning fear to context and fear to tone in rats, with the nonimmobilizers doing so at concentrations less than those that cause convulsions. (Fear was defined by freezing [volitional immobility] in the presence of the conditioned stimulus [context or tone].) However, the combination of isoflurane and 2N or flurothyl produced an antagonistic rather than an additive effect on learning, a finding in conflict with our hypothesis. And flurothyl was no less potent than 2N (at least no less potent relative to the concentration of each that produced convulsions) in its capacity to impair learning. We conclude that conventional inhaled anesthetics and nonimmobilizers impair learning and memory by different mechanisms. The basis for this impairment remains unknown. IMPLICATIONS: Conventional inhaled anesthetics and nonimmobilizers are antagonistic in their effects on learning and memory, and this finding suggests that they impair learning and memory, at least in part, by different mechanisms.