Basic Science of Epilepsy
Reducing premature KCC2 expression rescues seizure susceptibility and spine morphology in atypical febrile seizures
Awad PN, Sanon NT, Chattopadhyaya B, Carriço JN, Ouardouz M, Gagné J, Duss S, Wolf D, Desgent S, Cancedda L, Carmant L, Di Cristo G
Atypical febrile seizures are lateralized, prolonged and numerous; and can be a risk factor for epilepsy later in life. The link between cortical malformations, febrile seizures and epilepsy has been suggested, but the underlying mechanism isn’t fully understood. In this study, the scientists used a rat model (“LHS rats”) where cortical dysplasia and hyperthermia-induced seizures eventually lead to epilepsy and associated neurological sequelae. Given the role of KCC2 in GABAergic neurotransmission, they looked at expression and function of KCC2 in LHS rats. They found an increase in levels of KCC2 protein in these rats, a reduction in number of dendritic spines and abnormal dendritic maturation in area CA1 of the hippocampus. Reducing KCC2 protein expression using in-utero electroporation of shRNA reduced seizure number and corrected deficits in dendritic spines. Hence, alterations in KCC2 may be a common link between cortical dysplasia, febrile seizures and epilepsy.
GABAergic neurotransmission depends on KCC1 and NKCC2 Cl− co-transporters; a dysfunction in these co-transporters is seen in neonatal seizures. Bumetanide – a drug that blocks NKCC1 chloride cotransporters – has been shown to be beneficial in seizures in neonates. In the current study, the authors wanted to test whether bumetanide would be useful in status epilepticus (SE) by using brain slice electrophysiology and in vivo EEG experiments. Administration of bumetanide reduced seizure-like events induced by zero-Mg+², and in intact mice by administration of kainic acid. As seizures progressed, diazepam led to pharmacoresistance, which bumetanide was able to overcome. Hence, this study suggests that there might be a new indication of bumetanide in SE.
Marguet SL, Le-Schulte VT, Merseburg A, Neu A, Eichler R, Jakovcevski I, Ivanov A, Hanganu-Opatz IL, Bernard C, Morellini F, Isbrandt D.
There is a greater incidence of epilepsy during the first few years of life as neurodevelopmental processes are still underway. The current strategy is to administer anti-epileptic drugs after seizures occur. The authors of a recent paper investigated whether administration of bumetanide – a diuretic that blocks the NKCC1 cation-chloride co-transporter and dampens down depolarization – would be effective as an anti-epileptogenic agent in a mouse model of mutated Kv7 K+ channels. Transient administration of bumetanide was able to correct structural and functional changes in the hippocampus in the mutant mice. This study raises the possibility of bumetanide as an anti-epileptogenic agent.
Brain; Sept 2015
NMDAR antibody encephalitis is characterized by psychiatric symptoms like psychosis, paranoia and violent behavior, and may be followed by seizures. This study was done to examine the epileptogenicity of NMDAR antibodies in C57BL/6 mice. Administration of the antibody (IgG) into the lateral ventricle by itself did not cause spontaneous seizures, but exacerbated susceptibility of acute seizures caused by PTZ. Immunohistochemistry to visualize bound IgG showed that levels of IgG bound to the left hippocampus were proportional to seizure severity. This suggests that the IgG binds to, and causes internalization of NMDARs in the hippocampus, possibly leading to enhanced acutes eizures in response to PTZ.
Rettenbeck ML, von Rüden EL, Bienas S, Carlson R, Stein VM, Tipold A, Potschka H
Microglia are thought to contribute to epilepsy because they change shape and release reactive oxygen species (ROS). Most studies looking at microglial ROS production in epilepsy have been done after administration of a chemoconvulsant like pilocarpine or kainic acid. In the current study, the authors studied microglial ROS in an electrical status epilepticus (SE) model in rats. Using immunophenotyping to first identify microglia, the authors found an increase in microglial ROS two days after SE but not during epileptogenesis or epilepsy. Hence, anti-inflammatory strategies for epilepsy might be beneficial if they target the initial phase.
Shen HY, van Vliet EA, Bright KA, Hanthorn M, Lytle NK, Gorter J, Aronica E, Boison D
Glycine is an important modulator of synaptic plasticity in the hippocampus, but its role in epilepsy is not very well understood. In a recent study, the authors used two models of TLE – the intrahippocampal kainic acid model in mice and tetanic stimulation in rats, and observed effects on the glycine transporter 1 (GlyT1). In both rats and mice, the authors found an increase in protein levels of GlyT1. An increase in mRNA for GlyT1 was found in tissue resected from subjects with TLE. In a third experiment, mice with genetic deletion of GlyT1 in hippocampus and those administered the GlyT1 inhibitor LY showed an increased seizure threshold suggestive of a protective effect of decreasing glycine activity. Hence, the GlyT1 could be a potential novel target for development of new AEDs.
Astrogliosis (marked by hypertrophy of astrocytes and upregulation of astrocyte-specific markers) has been suggested to be proconvulsive in temporal lobe epilepsy. In this study, the authors show that there is loss of gap junction coupling between astrocytes in tissue resected from people with epilepsy with hippocampal sclerosis; this could underlie improper buffering of K+ ions. A loss of gap junction coupling between astrocytes was not noticed in tissue of patients who had epilepsy without hippocampal sclerosis. Using an animal model of epilepsy, the authors also show that astrocytic uncoupling is a fairly early process in epileptogenesis. Hence, one can hypothesize that halting astrocytic uncoupling after an initiating event could stop epileptogenesis; which is very exciting because at present, there is no such therapy that stops epileptogenesis in its tracks.
A novel anticonvulsant mechanism via inhibition of complement receptor C5ar1 in murine epilepsy model
Neurobiology of Disease 76(2015) 87-97
Studying inflammation could give us insights into novel drugs for seizure disorders. In this study, the researchers examined a certain pro-inflammatory complement peptide C5a and its receptor C5ar1 in experimental epilepsy. C5ar1 was found to be increased in experimental models of epilepsy; blockade of C5ar1 with an antagonist had anticonvulsant effects. Inhibition of C5ar1 decreased seizure-induced mortality and neurodegeneration. The anticonvulsant effects of C5ar1 inhibition were explained by a decrease in activation of the inflammatory mediator TNFα.
Targeting pharmacoresistant epilepsy and epileptogenesis with a dual-purpose antiepileptic drug
The authors of a recent study examined whether eslicarbazepine acetate (ESL) could target refractoriness to AEDs in addition to halting epileptogenesis. Using tissue resected from individuals with refractory epilepsy and pilocarpine- treated rodents, they found that ESL conferred use-dependent blockade of Na+ channels, hence alluding to its potential role in decreasing refractoriness. ESL was also found to block CaV3.2 calcium channels, mossy fiber sprouting and decrease neurodegeneration, hence having a likely beneficial role in slowing epileptogenesis.
Adenosine kinase, glutamine synthetase and EAAT2 as gene therapy targets for temporal lobe epilepsy
Young D, Fong DM, Lawlor PA, Wu A, Mouravlev A, McRae M, Glass M, Dragunow M, During MJ.
Anti-epileptic drugs (AEDs) work only via a few known targets; the authors of a recent study investigated whether astrocytes could prove to be a potential avenue for novel AED discovery. Adeno-associated viral (AAV) vector was used to overexpress glutamine synthetase (GS) and excitatory amino-acid transporter 2 (EAAT2) to increase synaptic glutamate clearance. Additionally, microRNA technique was used to decrease expression of the enzyme adenosine kinase (ADK), and increase synaptic levels of adenosine – a naturally occurring anticonvulsant. It was found that altering GS or EAAT2 did not affect seizures or neurodegeneration, but increasing adenosine levels did lead to a decrease in kainate-induced seizures and afforded neuroprotection.
Tau reduction prevents disease in a mouse model of Dravet syndrome
Ania L. Gheyara MD, PhD, Ravikumar Ponnusamy PhD, Biljana Djukic PhD, Ryan J. Craft BS, Kaitlyn Ho BS, Weikun Guo MS, Mariel M. Finucane PhD, Pascal E. Sanchez PhD, and Lennart Mucke MD
Objective: Reducing levels of the microtubule-associated protein tau has shown promise as a potential treatment strategy for diseases with secondary epileptic features such as Alzheimer disease. We wanted to determine whether tau reduction may also be of benefit in intractable genetic epilepsies.
Results: Tau ablation prevented the high mortality of Dravet mice and reduced the frequency of spontaneous and febrile seizures. It reduced interictal epileptic spikes in vivo and drug-induced epileptic activity in brain slices ex vivo. Tau ablation also prevented biochemical changes in the hippocampus indicative of epileptic activity and ameliorated abnormalities in learning and memory, nest building, and open field behaviors in Dravet mice. Deletion of only 1 Tau allele was sufficient to suppress epileptic activity and improve survival and nesting performance.
Interpretation: Tau reduction may be of therapeutic benefit in Dravet syndrome and other intractable genetic epilepsies.
Workshop on Neurobiology of Epilepsy appraisal: New systemic imaging technologies to study the brain in experimental models of epilepsy
Stefanie Dedeurwaerdere, Sandy R. Shultz, Paolo Federico, and Jerome Engel Jr
Successful application of functional neuroimaging of the whole brain in the animal laboratory now permits investigations during epileptogenesis and correlation with deep brain electroencephalography (EEG) activity. With the continuing development of these techniques and analytical methods, the potential for future translational research on epilepsy is enormous.
Losartan prevents acquired epilepsy via TGF-β signaling suppression
Guy Bar-Klein M.Med.Sc, Luisa P. Cacheaux Ph.D., Lyn Kamintsky M.Sc., Ofer Prager M.Med.Sc., Itai Weissberg M.Med.Sc., Karl Schoknecht M.D., Paul Cheng M.Sc., Soo Young Kim Ph.D., Lydia Wood Ph.D., Uwe Heinemann M.D., Ph.D., Daniela Kaufer Ph.D.2,, Alon Friedman M.D., Ph.D.
Objective: Acquired epilepsy is frequently associated with structural lesions following trauma, stroke and infections. While seizures are often difficult to treat, there is no clinically applicable strategy to prevent the development of epilepsy in patients at risk. We have recently shown that vascular injury is associated with activation of albumin-mediated transforming growth factor ÃŸ (TGF-ÃŸ) signaling, and followed by local inflammatory response and epileptiform activity ex vivo. Here we investigated albumin-mediated TGF-ÃŸ signaling and tested the efficacy of blocking the TGF-ÃŸ pathway in preventing epilepsy.
Encapsulated galanin-producing cells attenuate focal epileptic seizures in the hippocampus
Litsa Nikitidou, Malene Torp, Lone Fjord-Larsen, Philip Kusk, Lars U. Wahlberg, Mérab Kokaia
Our study shows that galanin-releasing encapsulated cell biodelivery (ECB) devices moderately suppress focal stimulation-induced recurrent seizures. Despite this moderate effect, the study provides conceptual proof that ECB could be a viable alternative approach to cell therapy in humans, with the advantage that the treatment could be terminated by removing these devices from the brain. Thereby, this strategy provides a higher level of safety for future therapeutic applications, in which genetically modified human cell lines that are optimized to produce and release antiepileptic compounds could be clinically evaluated for their seizure-suppressant effects.
Dravet syndrome patient-derived neurons suggest a novel epilepsy mechanism
Yu Liu MD, PhD, Luis F. Lopez-Santiago PhD, Yukun Yuan PhD, Julie M. Jones MS, Helen Zhang MS, Heather A. O'Malley PhD, Gustavo A. Patino PhD, Janelle E. O'Brien PhD, Raffaella Rusconi PhD, Ajay Gupta MD, Robert C. Thompson PhD, Marvin R. Natowicz MD, PhD, Miriam H. Meisler PhD, Lori L. Isom PhD, Jack M. Parent MD
Annals of Neurology, Volume 74, Issue 1, pages 128-139, July 2013.
Article first published online: 2 JUL 2013
Objective: Neuronal channelopathies cause brain disorders, including epilepsy, migraine, and ataxia. Despite the development of mouse models, pathophysiological mechanisms for these disorders remain uncertain. One particularly devastating channelopathy is Dravet syndrome (DS), a severe childhood epilepsy typically caused by de novo dominant mutations in the SCN1A gene encoding the voltage-gated sodium channel Nav1.1. Heterologous expression of mutant channels suggests loss of function, raising the quandary of how loss of sodium channels underlying action potentials produces hyperexcitability. Mouse model studies suggest that decreased Nav1.1 function in interneurons causes disinhibition. We aim to determine how mutant SCN1A affects human neurons using the induced pluripotent stem cell (iPSC) method to generate patient-specific neurons.
GABA progenitors grafted into the adult epileptic brain control seizures and abnormal behavior
Robert F Hunt, Kelly M Girskis, John L Rubenstein, Arturo Alvarez-Buylla, and Scott C Baraban
Impaired GABA-mediated neurotransmission has been implicated in many neurologic diseases, including epilepsy, intellectual disability and psychiatric disorders. We found that inhibitory neuron transplantation into the hippocampus of adult mice with confirmed epilepsy at the time of grafting markedly reduced the occurrence of electrographic seizures and restored behavioral deficits in spatial learning, hyperactivity and the aggressive response to handling.
Our results highlight a critical role for interneurons in epilepsy and suggest that interneuron cell transplantation is a powerful approach to halting seizures and rescuing accompanying deficits in severely epileptic mice.
Basic research using animal models
The April 2013 issue of Epilepsia contains a number of basic research papers using animal models. The following two papers reflect the power and opportunity for applying "bench work" to the real problems of clinical epileptology.
Ligia A. Papale, Christopher D. Makinson, J. Christopher Ehlen, Sergio Tufik, Michael J. Decker, Ketema N. Paul, Andrew Escayg
Gabi Dezsi, Ezgi Ozturk, Davor Stanic, Kim L. Powell, Hal Blumenfeld, Terence J. O'Brien, Nigel C. Jones
Canine epilepsy as a translational model?
Heidrun Potschka, Andrea Fischer, Eva-Lotta von Rüden, Velia Hülsmeyer, Wolfgang Baumgärtne
Dogs with spontaneous diseases can exhibit a striking similarity in etiology, clinical manifestation, and disease course when compared to human patients. Therefore, dogs are intensely discussed as a translational model of human disease. In particular, genetic studies in selected dog breeds serve as an excellent tool to identify epilepsy disease genes. The review gives an overview on the current state of knowledge regarding the etiology, clinical manifestation, pathology, and drug response of canine epilepsy, also pointing out the urgent need for further research on specific aspects. Moreover, the putative advantages, the disadvantages, and limitations of antiepileptic drug testing in canine epilepsy are critically discussed.
Detection of human papillomavirus in human focal cortical dysplasia type IIB
Annals of Neurology Vol 72, Issue 6, pp. 881-892. December 2012. Article first published online: 31 Dec 2012. DOI: 10.1002/ana.23795
Objective: Focal cortical dysplasia type IIB (FCDIIB) is a sporadic developmental malformation of the cerebral cortex highly associated with pediatric epilepsy. Balloon cells (BCs) in FCDIIB exhibit constitutive activation of the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Recently, the high-risk human papillomavirus type 16 oncoprotein E6 was identified as a potent activator of mTORC1 signaling. Here, we test the hypothesis that HPV16 E6 is present in human FCDIIB specimens.
Interpretation: Our results indicate a new association between HPV16 E6 and FCDIIB and demonstrate for the first time HPV16 E6 in the human brain. We propose a novel etiology for FCDIIB based on HPV16 E6 expression during fetal brain development. ANN NEUROL 2012;72:881-892
Eva M Jimenez-Mateos, Tobias Engel, Paula Merino-Serrais, Ross C McKiernan, Katsuhiro Tanaka, Genshin Mouri, Takanori Sano, Colm O'Tuathaigh, John L Waddington, Suzanne Prenter, Norman Delanty, Michael A Farrell, Donncha F O'Brien, RonÃ¡n M Conroy, Raymond L Stallings, Javier DeFelipe, David C Henshall
Abstract: Temporal lobe epilepsy is a common, chronic neurological disorder characterized by recurrent spontaneous seizures. MicroRNAs (miRNAs) are small, noncoding RNAs that regulate post-transcriptional expression of protein-coding mRNAs, which may have key roles in the pathogenesis of neurological disorders. In experimental models of prolonged, injurious seizures (status epilepticus) and in human epilepsy, we found upregulation of miR-134, a brain-specific, activity-regulated miRNA that has been implicated in the control of dendritic spine morphology. Silencing of miR-134 expression in vivo using antagomirs reduced hippocampal CA3 pyramidal neuron dendrite spine density by 21% and rendered mice refractory to seizures and hippocampal injury caused by status epilepticus. Depletion of miR-134 after status epilepticus in mice reduced the later occurrence of spontaneous seizures by over 90% and mitigated the attendant pathological features of temporal lobe epilepsy. Thus, silencing miR-134 exerts prolonged seizure-suppressant and neuroprotective actions; determining whether these are anticonvulsant effects or are truly antiepileptogenic effects requires additional experimentation. Read article
The editors of the historic textbook Jasper's Basic Mechanisms of Epilepsy announce the publication of the 4th edition by Oxford University Press. The book contains chapters from 90 international research groups summarizing the latest experimental advances in epilepsy for basic and clinical researchers.
This edition considers the role of interactions between neurons, synapses, and glia in the initiation, spread, and arrest of seizures. It examines mechanisms of excitability synchronization, and epileptogenesis and provides a framework for expanding the epilepsy genome and understanding complex heredity. It considers the mechanism of ion channelopathy, developmental epilepsy genes, and epilepsy comorbidities. And for the first time, it describes the current translation of epilepsy mechanisms into new therapeutic strategies.
Long considered the "bible" of basic epilepsy research, the volume provides encyclopedia coverage of current understanding and maps out new research directions for the coming decade.
Bianca Jupp, John Williams, David Binns, Rodney J. Hicks, Lisa Cardamone, Nigel Jones, Sandra Rees, Terence J. O'Brien
These findings demonstrate that hypometabolism occurs early in the processes of limbic epileptogenesis and is not merely a consequence of pyramidal cell loss or the progressive atrophy of limbic brain structures that follow. The hypometabolism may reflect cellular mechanisms occurring early during epileptogenesis in addition to any effects of the subsequent recurrent spontaneous seizures.
György Buzsáki, Costas A. Anastassiou, & Christof Koch
Neuronal activity in the brain gives rise to transmembrane currents that can be measured in the extracellular medium. Although the major contributor of the extracellular signal is the synaptic transmembrane current, other sources – including Na+ and Ca2+ spikes, ionic fluxes through voltage – and ligand-gated channels, and intrinsic membrane oscillations – can substantially shape the extracellular field. High-density recordings of field activity in animals and subdural grid recordings in humans, combined with recently developed data processing tools and computational modelling, can provide insight into the cooperative behaviour of neurons, their average synaptic input and their spiking output, and can increase our understanding of how these processes contribute to the extracellular signal.
Hierarchical clustering of brain activity during human nonrapid eye movement sleep
Mélanie Boly, Vincent Perlbarg, Guillaume Marrelec, Manuel Schabus, Steven Laureys, Julien Doyon, Mélanie Pélégrini-Issac, Pierre Maquet, and Habib Benali
Consciousness is reduced during nonrapid eye movement (NREM) sleep due to changes in brain function that are still poorly understood. Here, we tested the hypothesis that impaired consciousness during NREM sleep is associated with an increased modularity of brain activity.
Seizure-induced brain-born inflammation sustains seizure recurrence and blood-brain barrier damage
Laura Librizzi PhD, Francesco NoÃ¨ PhD, Annamaria Vezzani PhD, Marco de Curtis MD1, Teresa Ravizza PhD
Epilepsy is a common neurological disorder characterized by recurrent seizures often unresponsive to pharmacological treatment. Brain inflammation is considered a crucial etiopathogenetic mechanism of epilepsy that could be targeted to control seizures. Specific inflammatory mediators overexpressed in human epileptogenic foci are known to promote seizures in animal models. We investigate if seizures induce brain inflammation independently on extra-cerebral factors. We also verify whether brain-born inflammation is required and sufficient to maintain seizure activity and it supports blood-brain barrier (BBB) impairment. We addressed these questions by studying the relationship between seizures, inflammation and BBB permeability in a brain preparation isolated from extracerebral compartments.
Neuron-restrictive silencer factor-mediated hyperpolarization-activated cyclic nucleotide gated channelopathy in experimental temporal lobe epilepsy
McClelland S, Flynn C, Dubé C, Richichi C, Zha Q, Ghestem A, Esclapez M, Bernard C, Baram TZ. (2011)
Acquired HCN1 channelopathy derives from NRSF-mediated transcriptional repression that endures via chromatin modification and may provide insight into the mechanisms of a number of channelopathies that coexist with, and may contribute to, the conversion of a normal brain into an epileptic one.
Glutamatergic pre-ictal discharges emerge at the transition to seizure in human epilepsy
Gilles Huberfeld, Liset Menendez de la Prida, Johan Pallud, Ivan Cohen, Michel Le Van Quyen, Claude Adam, Stephane Clemenceau, Michel Baulac and Richard Miles
Brief and synchronous inter-ictal events can occur between seizures. Using human tissue samples and electroencephalography, this study shows that the transition from pre-ictal discharge to ictal discharge involves distinct temporal and spatial characteristics as well as glutamatergic mechanisms.
Single-neuron dynamics in human focal epilepsy
Wilson Truccolo, Jacob A Donoghue, Leigh R Hochberg, Emad N Eskandar, Joseph R Madsen, William S Anderson, Emery N Brown, Eric Halgren and Sydney S Cash
Epileptic seizures are traditionally considered to reflect hypersynchronous neuronal activity arising from runaway excitation. Here the authors analyze spike train patterns of single neurons during seizures in human epilepsy patients, finding that spiking activity during seizure initiation was highly heterogeneous in small cortical patches and across the network.
A Novel Positron Emission Tomography Imaging Protocol Identifies Seizure-Induced Regional Overactivity of P-Glycoprotein at the Blood-Brain Barrier.
Bankstahl JP, Bankstahl M, Kuntner C, Stanek J, Wanek T, Meier M, Ding XQ, MÃ¼ller M, Langer O, LÃ¶scher W.Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine, and Center for Systems Neuroscience, 30559 Hannover, Germany, Health and Environment Department, Molecular Medicine, AIT Austrian Institute of Technology, 2444 Seibersdorf, Austria, Department of Clinical Pharmacology, Medical University of Vienna, 1090 Vienna, Austria, and Department of Cardiology and Angiology and Institute of Diagnostic and Interventional Neuroradiology, Hannover Medical School, 30625 Hannover, Germany.
Exome sequencing of ion channel genes reveals complex profiles confounding personal risk assessment in epilepsy.
Klassen T, Davis C, Goldman A, Burgess D, Chen T, Wheeler D, McPherson J, Bourquin T, Lewis L, Villasana D, Morgan M, Muzny D, Gibbs R, Noebels J.