Temporal Coordination of Hippocampal Neurons Reflects Cognitive Outcome Post-febrile Status Epilepticus

AbstractThe coordination of dynamic neural activity within and between neural networks is believed to underlie normal cognitive processes. Conversely, cognitive deficits that occur following neurological insults may result from network discoordination. We hypothesized that cognitive outcome following febrile status epilepticus (FSE) depends on network efficacy within and between fields CA1 and CA3 to dynamically organize cell activity by theta phase. Control and FSE rats were trained to forage or perform an active avoidance spatial task. FSE rats were sorted by those that were able to reach task criterion (FSE-L) and those that could not (FSE-NL). FSE-NL CA1 place cells did not exhibit phase preference in either context and exhibited poor cross-theta interaction between CA1 and CA3. FSE-L and control CA1 place cells exhibited phase preference at peak theta that shifted during active avoidance to the same static phase preference observed in CA3. Temporal coordination of neuronal activity by theta phase may therefore explain variability in cognitive outcome following neurological insults in early development

Pharmacological study of T-type calcium channels in mice models of convulsion and epileptogenesis

De nombreuses études expérimentales montrent que les canaux calciques activés par la dépolarisation membranaire, tout particulièrement les canaux calciques de type T (canaux T), jouent un rôle important dans la physiopathologie des épilepsies. Il existe trois isoformes des canaux T, Cav3.1, Cav3.2 et Cav3.3, toutes exprimées au niveau neuronal. De manière classique, c'est dans l'épilepsie absence où les canaux T ont été le plus étudiés. Les canaux T jouent également un rôle dans des modèles d'épilepsie partielle secondairement généralisée, comme le modèle pilocarpine qui mime l'épilepsie du lobe temporal (ELT). Jusqu'à présent ces canaux ne possédaient pas de pharmacologie spécifique, mais plusieurs molécules récemment synthétisées, en particulier le TTA-A2, apparaissent sélectives des canaux T. Le premier objectif de ma thèse était d'étudier l'implication des canaux T dans l'épileptogenèse. Pour cela nous avons traité des souris au TTA-A2 pendant la phase de latence du modèle pilocarpine (modèle ELT). Nos conditions expérimentales ne nous ont pas permis de conclure quant à une action protectrice du TTA-A2 dans ce modèle. Le deuxième objectif était d'étudier l'effet du TTA-A2 sur des modèles murins de convulsions généralisées : le modèle du Maximal Electroshock Seizure (MES) et le modèle pentylènetétrazole (PTZ). Deux lignées de souris inactivées pour les isoformes Cav3.1 ou Cav3.2 (KO Cav3.1 et KO Cav3.2) ont également été caractérisées dans cette étude. Nous montrons que le TTA-A2 réduit l'apparition des crises toniques dans le modèle MES et que les souris KO Cav3.1 sont également protégées, suggérant un rôle prépondérant des canaux Cav3.1 dans le développement des crises toniques.Numerous experimental studies show that calcium channels activated by membrane depolarization, especially T-type calcium channels (T-channels), play an important role in the physiopathology of epilepsy. There are three T-channels isoforms, Cav3.1, Cav3.2 and Cav3.3, all expressed in neuronal level. Conventionally, T-channels were the most studied in absence epilepsy. T-channels are also involved in partial secondarily generalized epilepsy models, as the pilocarpine model that mimics temporal lobe epilepsy (TLE).Up to now, there was no specific pharmacology for this channels, but several molecules have recently been synthesized, particularly TTA-A2, appearing selective T-channels. The first goal of my thesis was to study the T-channels involvement in epileptogenesis. For this purpose we treated mice with TTA-A2 during the silent phase of the pilocarpine model (TLE model). Our experimental conditions do not allow us to conclude about a possible protective action of TTA-A2 on this model. The second goal was to study TTA-A2 effects on mice models of generalized seizures: the Maximal Electroshock model (MES) and the pentylenetetrazole model (PTZ). Two mice strains knock-out for Cav3.1 or Cav3.2 (KO Cav3.1 and KO Cav3.2) have also been characterized in this study. We show that the TTA-A2 reduces the appearance of tonic seizures in the MES model and the KO Cav3.1 mice are also protected, suggesting a preponderant role of Cav3.1 channels in the development of tonic seizures

Etude pharmacologique des canaux calciques de type T dans des modèles murins de convulsion et d'épileptogenèse.

De nombreuses études expérimentales montrent que les canaux calciques activés par la dépolarisation membranaire, tout particulièrement les canaux calciques de type T (canaux T), jouent un rôle important dans la physiopathologie des épilepsies. Il existe trois isoformes des canaux T, Cav3.1, Cav3.2 et Cav3.3, toutes exprimées au niveau neuronal. De manière classique, c'est dans l'épilepsie absence où les canaux T ont été le plus étudiés. Les canaux T jouent également un rôle dans des modèles d'épilepsie partielle secondairement généralisée, comme le modèle pilocarpine qui mime l'épilepsie du lobe temporal (ELT). Jusqu'à présent ces canaux ne possédaient pas de pharmacologie spécifique, mais plusieurs molécules récemment synthétisées, en particulier le TTA-A2, apparaissent sélectives des canaux T. Le premier objectif de ma thèse était d'étudier l'implication des canaux T dans l'épileptogenèse. Pour cela nous avons traité des souris au TTA-A2 pendant la phase de latence du modèle pilocarpine (modèle ELT). Nos conditions expérimentales ne nous ont pas permis de conclure quant à une action protectrice du TTA-A2 dans ce modèle. Le deuxième objectif était d'étudier l'effet du TTA-A2 sur des modèles murins de convulsions généralisées : le modèle du Maximal Electroshock Seizure (MES) et le modèle pentylènetétrazole (PTZ). Deux lignées de souris inactivées pour les isoformes Cav3.1 ou Cav3.2 (KO Cav3.1 et KO Cav3.2) ont également été caractérisées dans cette étude. Nous montrons que le TTA-A2 réduit l'apparition des crises toniques dans le modèle MES et que les souris KO Cav3.1 sont également protégées, suggérant un rôle prépondérant des canaux Cav3.1 dans le développement des crises toniques.Numerous experimental studies show that calcium channels activated by membrane depolarization, especially T-type calcium channels (T-channels), play an important role in the physiopathology of epilepsy. There are three T-channels isoforms, Cav3.1, Cav3.2 and Cav3.3, all expressed in neuronal level. Conventionally, T-channels were the most studied in absence epilepsy. T-channels are also involved in partial secondarily generalized epilepsy models, as the pilocarpine model that mimics temporal lobe epilepsy (TLE).Up to now, there was no specific pharmacology for this channels, but several molecules have recently been synthesized, particularly TTA-A2, appearing selective T-channels. The first goal of my thesis was to study the T-channels involvement in epileptogenesis. For this purpose we treated mice with TTA-A2 during the silent phase of the pilocarpine model (TLE model). Our experimental conditions do not allow us to conclude about a possible protective action of TTA-A2 on this model. The second goal was to study TTA-A2 effects on mice models of generalized seizures: the Maximal Electroshock model (MES) and the pentylenetetrazole model (PTZ). Two mice strains knock-out for Cav3.1 or Cav3.2 (KO Cav3.1 and KO Cav3.2) have also been characterized in this study. We show that the TTA-A2 reduces the appearance of tonic seizures in the MES model and the KO Cav3.1 mice are also protected, suggesting a preponderant role of Cav3.1 channels in the development of tonic seizures.MONTPELLIER-BU Médecine UPM (341722108) / SudocSudocFranceF

Mouse hippocampal phosphorylation footprint induced by generalized seizures: Focus on ERK, mTORC1 and Akt/GSK-3 pathways

International audienceExacerbated hippocampal activity has been associated to critical modifications of the intracellular signaling pathways. We have investigated rapid hippocampal adaptive responses induced by maximal electroshock seizure (MES). Here, we demonstrate that abnormal and exacerbated hippocampal activity induced by MES triggers specific and temporally distinct patterns of phosphorylation of extracellular signal-related kinase (ERK), mammalian target of rapamycin complex (mTORC) and Akt/glycogen synthase kinase-3 (Akt/GSK-3) pathways in the mouse hippocampus. While the ERK pathway is transiently activated, the mTORC1 cascade follows a rapid inhibition followed by a transient activation. This rebound of mTORC1 activity leads to the selective phosphorylation of p70S6K, which is accompanied by an enhanced phosphorylation of the ribosomal subunit S6. In contrast, the Akt/GSK-3 pathway is weakly altered. Finally, MES triggers a rapid upregulation of several plasticity-associated genes as a consequence exacerbated hippocampal activity. The results reported in the present study are reminiscent of the one observed in other models of generalized seizures, thus defining a common molecular footprint induced by intense and aberrant hippocampal activities

Focal Dorsal Hippocampal Nav1.1 Knock Down Alters Place Cell Temporal Coordination and Spatial Behavior

International audienceAlterations in the voltage-gated sodium channel Nav.1.1 are implicated in various neurological disorders, including epilepsy, Alzheimer's disease, and autism spectrum disorders. Previous studies suggest that the reduction of Nav1.1 expression leads to a decrease of fast spiking activity in inhibitory neurons. Because interneurons (INs) play a critical role in the temporal organization of neuronal discharge, we hypothesize that Nav1.1 dysfunction will negatively impact neuronal coordination in vivo. Using shRNA interference, we induced a focal Nav1.1 knock-down (KD) in the dorsal region of the right hippocampus of adult rats. Focal, unilateral Nav1.1 KD decreases the performance in a spatial novelty recognition task and the firing rate in INs, but not in pyramidal cells. It reduced theta/gamma coupling of hippocampal oscillations and induced a shift in pyramidal cell theta phase preference. Nav1.1 KD degraded spatial accuracy and temporal coding properties of place cells, such as theta phase precession and compression of ongoing sequences. Aken together, these data demonstrate that a deficit in Nav1.1 alters the temporal coordination of neuronal firing in CA1 and impairs behaviors that rely on the integrity of this network. They highlight the potential contribution of local inhibition in neuronal coordination and its impact on behavior in pathological conditions

Blockade of T-type calcium channels prevents tonic-clonic seizures in a maximal electroshock seizure model

International audienc

Data_Sheet_1_The Shank3Venus/Venus knock in mouse enables isoform-specific functional studies of Shank3a.pdf

BackgroundShank3 is a scaffolding protein essential for the organization and function of the glutamatergic postsynapse. Monogenic mutations in SHANK3 gene are among the leading genetic causes of Autism Spectrum Disorders (ASD). The multiplicity of Shank3 isoforms seems to generate as much functional diversity and yet, there are no tools to study endogenous Shank3 proteins in an isoform-specific manner.MethodsIn this study, we created a novel transgenic mouse line, the Shank3Venus/Venus knock in mouse, which allows to monitor the endogenous expression of the major Shank3 isoform in the brain, the full-length Shank3a isoform.ResultsWe show that the endogenous Venus-Shank3a protein is localized in spines and is mainly expressed in the striatum, hippocampus and cortex of the developing and adult brain. We show that Shank3Venus/+ and Shank3Venus/Venus mice have no behavioral deficiency. We further crossed Shank3Venus/Venus mice with Shank3ΔC/ΔC mice, a model of ASD, to track the Venus-tagged wild-type copy of Shank3a in physiological (Shank3Venus/+) and pathological (Shank3Venus/ΔC) conditions. We report a developmental delay in brain expression of the Venus-Shank3a isoform in Shank3Venus/ΔC mice, compared to Shank3Venus/+ control mice.ConclusionAltogether, our results show that the Shank3Venus/Venus mouse line is a powerful tool to study endogenous Shank3a expression, in physiological conditions and in ASD.</p

The mGlu7 receptor provides protective effects against epileptogenesis and epileptic seizures

International audienceFinding new targets to control or reduce seizure activity is essential to improve the management of epileptic patients. We hypothesized that activation of the pre-synaptic and inhibitory metabotropic glutamate receptor type 7 (mGlu7) reduces spontaneous seizures. We tested LSP2-9166, a recently developed mGlu7/4 agonist with unprecedented potency on mGlu7 receptors, in two paradigms of epileptogenesis. In a model of chemically induced epileptogenesis (pentylenetetrazole systemic injection), LSP2-9166 induces an anti-epileptogenic effect rarely observed in preclinical studies. In particular, we found a bidirectional modulation of seizure progression by mGlu4 and mGlu7 receptors, the latter preventing kindling. In the intra-hippocampal injection of kainic acid mouse model that mimics the human mesial temporal lobe epilepsy, we found that LSP2-9166 reduces seizure frequency and hippocampal sclerosis. LSP2-9166 also acts as an anti-seizure drug on established seizures in both models tested. Specific modulation of the mGlu7 receptor could represent a novel approach to reduce pathological network remodeling

Restoring glutamate receptosome dynamics at synapses rescues autism-like deficits in Shank3-deficient mice

International audienceShank3 monogenic mutations lead to autism spectrum disorders (ASD). Shank3 is part of the glutamate receptosome that physically links ionotropic NMDA receptors to metabotropic mGlu5 receptors through interactions with scaffolding proteins PSD95-GKAP-Shank3-Homer. A main physiological function of the glutamate receptosome is to control NMDA synaptic function that is required for plasticity induction. Intact glutamate receptosome supports glutamate receptors activation and plasticity induction, while glutamate receptosome disruption blocks receptors activity, preventing the induction of subsequent plasticity. Despite possible impact on metaplasticity and cognitive behaviors, scaffold interaction dynamics and their consequences are poorly defined. Here, we used mGlu5-Homer interaction as a biosensor of glutamate receptosome integrity to report changes in synapse availability for plasticity induction. Combining BRET imaging and electrophysiology, we show that a transient neuronal depolarization inducing NMDA-dependent plasticity disrupts glutamate receptosome in a long-lasting manner at synapses and activates signaling pathways required for the expression of the initiated neuronal plasticity, such as ERK and mTOR pathways. Glutamate receptosome disruption also decreases the NMDA/AMPA ratio, freezing the sensitivity of the synapse to subsequent changes of neuronal activity. These data show the importance of a fine-tuning of protein-protein interactions within glutamate receptosome, driven by changes of neuronal activity, to control plasticity. In a mouse model of ASD, a truncated mutant form of Shank3 prevents the integrity of the glutamate receptosome. These mice display altered plasticity, anxiety-like, and stereotyped behaviors. Interestingly, repairing the integrity of glutamate receptosome and its sensitivity to the neuronal activity rescued synaptic transmission, plasticity, and some behavioral traits of Shank3∆C mice. Altogether, our findings characterize mechanisms by which Shank3 mutations cause ASD and highlight scaffold dynamics as new therapeutic target