Lee S. Stewart

Hippocampal Melatonin Receptors Modulate Seizure Threshold

Summary:u2002 Purpose: The pineal hormone melatonin has been shown to enhance hippocampal excitability. We therefore investigated whether inactivation of hippocampal melatonin receptors affects behavioral seizures.

Temporal lobe seizures alter the amplitude and timing of rat behavioral rhythms.

Daily rhythms of spontaneous locomotor activity (SLA) in rats were studied before and after an episode of pilocarpine-induced convulsive status epilepticus (SE). A pronounced increase in activity levels during both the light and dark phases was found 1 week after SE as compared with baseline SLA and controls administered saline. Rats with bilateral lesions of the nucleus accumbens (shell) did not exhibit any significant change in SLA 1 week after SE compared with controls. We suggest that during the first week after SE the increase in SLA was induced by abnormal neuronal activity in the hippocampus driving a descending limbic-motor pathway via the nucleus accumbens. EEG recordings revealed high-amplitude interictal spikes in hippocampal CA1. During subsequent weeks, SLA rhythms of nonlesioned chronic epileptic rats remained elevated but progressively normalized over a period of 12 weeks. Although both chronic epileptic and control groups displayed near-24-hour activity patterns under light-dark conditions, significant delays (>4 hour) in acrophase were observed after spontaneous seizures had developed. The phase delay was positively correlated with seizure history and likely the result of postictal hyperactivity associated with seizures during the normal rest period. Despite these changes, cell density in the suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL) did not differ significantly between epileptic and control groups. In the absence of damage to brain areas directly involved with the regulation of behavioral rhythms, chronic seizure activity presumably alters the timing of activity patterns through a nonphotic mechanism, perhaps involving activation of the SCN or IGL during limbic seizures.

Diurnal variation in pilocarpine-induced generalized tonic–clonic seizure activity

The efficacy of the cholinergic agonist pilocarpine to evoke generalized seizures in rats was examined over the 24-h photocycle. Both seizure latency and severity during the 24-h period exhibited marked diurnal variation. Seizure activity generalized more rapidly throughout the dark phase and was often characterized by wild running clonus and tonic extensor convulsions. In contrast, electrographic seizures during the light phase were mainly associated with facial/forelimb clonus and rearing typical of pilocarpine-induced seizures. These data suggest that the sensitivity of the rodent brain to pilocarpine exhibits day-night variation and seizure activity induced during the dark phase may generalize via the brainstem.

Severity of atypical absence phenotype in GABAB transgenic mice is subunit specific.

Overexpression of GABA(B)R1a receptors in mice (R1a(+)) results in an atypical absence seizure phenotype characterized by 3- to 6-Hz slow spike-and-wave discharges (SSWDs), reduced synaptic plasticity, and cognitive impairment. Here we tested the hypothesis that increased R1 expression causes atypical absence epilepsy and is not subunit specific. GABA(B)R1b receptors were overexpressed in mouse forebrain (R1b(+)) and confirmed by immunoblot and (3)H-CGP54626A autoradiography. The R1b(+) mice showed a reduction in hippocampal long-term potentiation and GABA(A) receptor-mediated inhibitory postsynaptic currents. R1b(+) mice manifested an electrographic, pharmacological, and behavioral phenotype consistent with atypical absence seizures, though less robust than R1a(+) in terms of SSWD duration and severity of cognitive impairment. These results suggest that abnormal GABA(B)R1b function plays a lesser role in the development of atypical absence epilepsy.

Endogenous melatonin and epileptogenesis : Facts and hypothesis

In human epilepsy, diurnal variation in seizure phenomena suggests the involvement of a time-dependent biological signal. Clinical evidence indicates that in some cases, temporal clustering of epileptic seizures is in phase with the nocturnal rise in circulating melatonin. Although this hormone has been reported to stabilize the brain against seizure-producing stimuli, these pharmacological doses are not representative of physiological conditions but would nonetheless facilitate widespread inhibitory neurotransmission characteristic of traditional anticonvulsants. Instead, it is proposed that endogenous melatonin contributes to epileptiform activity through inhibitory actions on dopaminergic activity. Dopamine is considered a natural downregulator of seizure activity in a number of species, including humans, and numerous lines of evidence suggest that melatonin is capable of stimulating a decrease in dopamine output within areas of the brain thought to participate in the control of epileptic seizures. Pharmacological manipulation of the endogenous melatonin rhythm may provide a useful therapeutic strategy against the occurrence of seizures during increased hormone production.

Circadian distribution of generalized tonic-clonic seizures associated with murine succinic semialdehyde dehydrogenase deficiency, a disorder of GABA metabolism

Human succinic semialdehyde dehydrogenase (SSADH) deficiency is an autosomal recessive disorder of GABA metabolism associated with motor impairment and epileptic seizures. Similarly, mice with targeted deletion of the Aldh5a1 gene (Aldh5a1(-/-)) exhibit SSADH deficiency and seizures early in life. These seizures begin as absence seizures the second week of life, but evolve into generalized convulsive seizures that increase in severity and become lethal during the fourth postnatal week. The seizures are alleviated and survival is prolonged when the mutant animals are weaned onto a ketogenic diet (KD). The persistence of spontaneous, recurrent, generalized tonic-clonic seizures in KD-treated adult Aldh5a1(-/-) mice allowed us to quantify their daily (circadian) distribution using a novel behavioral method based on the detection of changes in movement velocity. Adult KD-treated Aldh5a1(-/-) mice exhibited a seizure phenotype characterized by fits of wild running clonus accompanied by jumping and bouncing. These hypermotor seizures were largely spontaneous and occurred daily in a nonrandom pattern. The seizure rhythm showed a peak shortly after dark phase onset (2008 hours) with near-24-hour periodicity. Age-matched wild-type littermates showed no evidence of abnormal motor behavior. These new data suggest that generalized tonic-clonic seizures in Aldh5a1(-/-) mice are more frequent during a specific time of day and will provide useful information to clinicians for the treatment of seizures associated with human SSADH deficiency.

Environmental Enrichment Improves Behavioral Outcome in the AY-9944 Model of Childhood Atypical Absence Epilepsy

ABSTRACT Atypical absence seizures are drug resistant in the majority of children with Lennox–Gastaut syndrome and herald a poor neurodevelopmental outcome. Here we studied the effects of environmental enrichment, enriched housing conditions designed to stimulate sensory and motor systems in the brain, on behavioral outcome in mice treated with the cholesterol biosynthesis inhibitor AY-9944 (AY), a clinically relevant model of atypical absence epilepsy. Beginning at postnatal day (P) 2, C3H mice were treated with AY (7.5 mg/kg) every 6 days until P20 and then weaned into enriched or standard cages. After 30 days (∼P50), AY mice from the enriched housing condition exhibited less behavioral hyperactivity and anxiety, improved olfactory recognition, and spatial learning, but no significant reduction in the number of ictal discharges in comparison with their non-enriched cohorts. The beneficial effects of environmental enrichment in AY model were in some behavioral tests gender-specific in favor of males suggesting that other, possibly hormonally mediated mechanisms, may interact with the therapeutic effects of enrichment. Taken together, these data provide a starting point to derive clinical occupational therapies for improving behavioral outcome in cases of intractable childhood seizures.

Pilocarpine model of temporal lobe epilepsy shows enhanced response to general anesthetics

Complex partial seizures, commonly arising from temporal lobe epilepsy (TLE), are associated with neuronal loss and post-seizure impairment of consciousness. We tested the hypothesis that TLE subjects, in between seizures, are associated with a decreased level of consciousness that is manifested by an enhanced response to a general anesthetic. Two animal models of TLE--amygdala kindling and pilocarpine-induced status epilepticus (Pilo-SE)--were tested. Pilo-SE rats, but not amygdala-kindled rats, showed a prolonged loss of pain and righting responses after 20 and 40 mg/kg i.p. pentobarbital, 2% halothane, and 5 and 10 mg/kg i.v. propofol as compared to control saline-treated rats. Since the major pathology of Pilo-SE rats was cell loss in the piriform cortex (PC) and the entorhinal cortex (EC), we studied the anesthetic response after inactivation of the EC or PC by locally infusing GABAA receptor agonist muscimol. Muscimol inactivation of the PC or EC, as compared to saline infusion in the same rats, prolonged the duration of loss of righting reflex, typically without changing the duration of loss of tail-pinch response, after 20 mg/kg i.p. pentobarbital, 2% halothane and 5 mg/kg i.v. propofol. Muscimol infusion, as compared to saline infusion, in the PC or EC also tended to decrease 30-100 Hz gamma EEG in the frontal cortex. In conclusion, a TLE model that resulted in neuronal loss, Pilo-SE, enhanced the response to a general anesthetic that could partly be attributed to a loss of neurons in the EC and PC.

Large differences in blood measures, tissue weights, and focal areas of damage 1 year after postseizure treatment with acepromazine or ketamine

Approximately 1 year after rats were seized as young adults with lithium (3 mEq/kg) and pilocarpine (30 mg/kg) and given acepromazine or ketamine, 18 blood measures, wet tissue weights, and detailed damage scores for 107 brain structures were completed. Compared with normal and ketamine-treated rats, acepromazine-treated seized rats (total n=54) had lighter pancreata and spleens and elevated aspartate aminotransferase and alanine aminotransferase blood levels. Even though average damage did not differ, the mosaic of brain damage completely discriminated the two seized groups. Differential effects of postseizure treatment on functions of the thyroid, pancreas, and spleen were indicated. Ketamine-treated seized rats were healthier than acepromazine-treated seized rats or normal rats. This experiment demonstrates the importance of whole-organism assessment and that the single administration of a specific drug after onset of status epilepticus can produce marked differences in the evolution of brain damage and its influence on specific organs for the rest of the animals life.

Corrigendum to “Large differences in blood measures, tissue weights, and focal areas of damage 1 year after postseizure treatment with acepromazine or ketamine” [Epilepsy & Behavior 15, Issue 2 (2009) 98-105]

a Department of Biology, Laurentian University, 935 Ramsey Lake Road, Sudbury, ON, Canada b Behavioural Neuroscience Program, Laurentian University, Sudbury, ON, Canada c School of Health Sciences and Emergency Services, Cambrian College of Applied Arts and Technology, Sudbury, ON, Canada d Neuroscience and Mental Health Research Program, The Hospital for Sick Children, Toronto, ON, Canada e Biomolecular Sciences Program, Laurentian University, Sudbury, ON, Canada f Northeast Mental Health Centre, North Bay, ON, Canada