RESUMEN
BACKGROUND: Cortical ischemic lesions represent the predominant pathomorphological pattern of focal lesions after aneurysmal subarachnoid hemorrhage (aSAH). Autopsy studies suggest that they occur adjacent to subarachnoid blood and are related to spasm of small cortical rather than proximal arteries. Recent clinical monitoring studies showed that cortical spreading depolarizations, which induce cortical arterial spasms, are involved in lesion development. If subarachnoid blood induces adjacent cortical lesions, it would be expected that (i) they also develop after traumatic subarachnoid hemorrhage (tSAH), and (ii) lesions after tSAH can occur in absence of angiographic vasospasm, as was found for aSAH. CASE PRESENTATION: An 86-year-old woman was admitted to our hospital with fluctuating consciousness after hitting her head during a fall. The initial computed tomography (CT) was significant for tSAH in cortical sulci. On day 8, the patient experienced a secondary neurological deterioration with reduced consciousness and global aphasia. Whereas the CT scan on day 9 was still unremarkable, magnetic resonance imaging (MRI) on day 10 revealed new cortical laminar infarcts adjacent to sulcal blood clots. Proximal vasospasm was ruled out using MR and CT angiography and Doppler sonography. CT on day 14 confirmed the delayed infarcts. CONCLUSIONS: We describe a case of delayed cortical infarcts around sulcal blood clots after tSAH in the absence of proximal vasospasm, similar to results found previously for aSAH. As for aSAH, this case suggests that assessment of angiographic vasospasm is not sufficient to screen for risk of delayed infarcts after tSAH. Electrocorticography is suggested as a complementary method to monitor the hypothesized mechanism of spreading depolarizations.
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Infarto Cerebral/diagnóstico , Infarto Cerebral/etiología , Hemorragia Subaracnoidea Traumática/complicaciones , Anciano de 80 o más Años , Angiografía por Tomografía Computarizada , Electrocorticografía , Femenino , Humanos , Imagen por Resonancia Magnética , Trombosis/complicaciones , Tomografía Computarizada por Rayos X , Vasoespasmo IntracranealRESUMEN
Neuropixels probes have become a crucial tool for high-density electrophysiological recordings. Although most research involving these probes is in acute preparations, some scientific inquiries require long-term recordings in freely moving animals. Recent reports have presented prosthesis designs for chronic recordings, but some of them do not allow for probe recovery, which is desirable given their cost. Others appear to be fragile, as these articles describe numerous broken implants.Objective.This fragility presents a challenge for recordings in rats, particularly in epilepsy models where strong mechanical stress impinges upon the prosthesis. To overcome these limitations, we sought to develop a new prosthesis for long-term electrophysiological recordings in healthy and epileptic rats.Approach.We present a new prosthesis specifically designed to protect the probes from strong shocks and enable the safe retrieval of probes after experiments.Main results.This prosthesis was successfully used to record from healthy and epileptic rats for up to three weeks almost continuously. Overall, 10 out of 11 probes could be successfully retrieved with a retrieval and reuse success rate of 91%.Significance.Our design and protocol significantly improved previously described probe recycling performances and prove usage on epileptic rats.
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Miembros Artificiales , Epilepsia , Ratas , Animales , Electrodos Implantados , Fenómenos Electrofisiológicos , PolímerosRESUMEN
BACKGROUND AND PURPOSE: The KCNQ2 gene encodes for the Kv 7.2 subunit of non-inactivating potassium channels. KCNQ2-related diseases range from autosomal dominant neonatal self-limited epilepsy, often caused by KCNQ2 haploinsufficiency, to severe encephalopathies caused by KCNQ2 missense variants. In vivo and in vitro effects of the sodium channel blocker eslicarbazepine acetate (ESL) and eslicarbazepine metabolite (S-Lic) in a mouse model of self-limited neonatal epilepsy as a first attempt to assess the utility of ESL in the KCNQ2 disease spectrum was investigated. EXPERIMENTAL APPROACH: Effects of S-Lic on in vitro physiological and pathological hippocampal neuronal activity in slices from mice carrying a heterozygous deletion of Kcnq2 (Kcnq2+/- ) and Kcnq2+/+ mice were investigated. ESL in vivo efficacy was investigated in the 6-Hz psychomotor seizure model in both Kcnq2+/- and Kcnq2+/+ mice. KEY RESULTS: S-Lic increased the amplitude and decreased the incidence of physiological sharp wave-ripples in a concentration-dependent manner and slightly decreased gamma oscillations frequency. 4-Aminopyridine-evoked seizure-like events were blocked at high S-Lic concentrations and substantially reduced in incidence at lower concentrations. These results were not different in Kcnq2+/+ and Kcnq2+/- mice, although the EC50 estimation implicated higher efficacy in Kcnq2+/- animals. In vivo, Kcnq2+/- mice had a lower seizure threshold than Kcnq2+/+ mice. In both genotypes, ESL dose-dependently displayed protection against seizures. CONCLUSIONS AND IMPLICATIONS: S-Lic slightly modulates hippocampal oscillations and blocks epileptic activity in vitro and in vivo. Our results suggest that the increased excitability in Kcnq2+/- mice is effectively targeted by S-Lic high concentrations, presumably by blocking diverse sodium channel subtypes.
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Epilepsia del Lóbulo Temporal , Epilepsia , Animales , Anticonvulsivantes/farmacología , Anticonvulsivantes/uso terapéutico , Dibenzazepinas , Epilepsia/tratamiento farmacológico , Epilepsia del Lóbulo Temporal/tratamiento farmacológico , Canal de Potasio KCNQ2/genética , Canal de Potasio KCNQ2/metabolismo , Ratones , Convulsiones/tratamiento farmacológicoRESUMEN
The subiculum (Sub) is the principal target of CA1 pyramidal cells. It serves as the final relay of hippocampal output and thus mediates hippocampal-cortical interaction. In addition, the Sub receives direct input from the entorhinal cortex via the temporoammonic pathway. In this study, we demonstrate that low-frequency stimulation of the temporoammonic pathway results in the disinhibition of excitatory synaptic transmission at CA1-Sub synapses. We provide evidence that this disinhibition is mediated by an NMDA receptor-dependent long-term depression (LTD) of GABAergic inhibition. This mechanism might bear physiological significance for the stabilization and processing of mnemonic information at hippocampal output synapses and underpins the functional role of hippocampal-entorhinal interaction in memory formation.
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Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/fisiología , Plasticidad Neuronal/fisiología , Sinapsis/fisiología , Animales , Región CA1 Hipocampal/fisiología , Estimulación Eléctrica/métodos , Masculino , Vías Nerviosas/fisiología , Ratas , Ratas Wistar , Transmisión Sináptica/fisiologíaRESUMEN
Epileptiform activity alters gene expression in the central nervous system, a phenomenon that has been studied extensively in animal models. Here, we asked whether also in vitro models of seizures are in principle suitable to investigate changes in gene expression due to epileptiform activity and tested this hypothesis mainly in rodent and additionally in some human brain slices. We focused on three genes relevant for seizures and epilepsy: FOS proto-oncogene (c-Fos), inducible cAMP early repressor (Icer) and mammalian target of rapamycin (mTor). Seizure-like events (SLEs) were induced by 4-aminopyridine (4-AP) in rat entorhinal-hippocampal slices and by 4-AP/8 mM potassium in human temporal lobe slices obtained from surgical treatment of epilepsy. SLEs were monitored simultaneously by extracellular field potentials and intrinsic optical signals (IOS) for 1-4 h, mRNA expression was quantified by real time PCR. In rat slices, both duration of SLE exposure and SLE onset region were associated with increased expression of c-Fos and Icer while no such association was shown for mTor expression. Similar to rat slices, c-FOS induction in human tissue was increased in slices with epileptiform activity. Our results indicate that irrespective of limitations imposed by ex vivo conditions, in vitro models represent a suitable tool to investigate gene expression. Our finding is of relevance for the investigation of human tissue that can only be performed ex vivo. Specifically, it presents an important prerequisite for future studies on transcriptome-wide and cell-specific changes in human tissue with the goal to reveal novel candidates involved in the pathophysiology of epilepsy and possibly other CNS pathologies.
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Malformations of cortical development (MCD) represent a group of rare diseases with severe clinical presentation as epileptic and pharmacoresistant encephalopathies. Morphological studies in tissue from MCD patients have revealed reduced GABAergic efficacy and increased intracellular chloride concentration in neuronal cells as important pathophysiological mechanisms in MCD. Also, in various animal models, alterations of GABAergic inhibition have been postulated as a predominant factor contributing to perilesional hyperexcitability. Along with this line, the NKCC1 inhibitor bumetanide has been postulated as a potential drug for treatment of epilepsy, mediating its antiepileptic effect by reduction of the intracellular chloride and increased inhibitory efficacy of GABAergic transmission. In the present study, we focused on the focal freeze-lesion model of MCD to compare antiepileptic drugs with distinct mechanisms of action, including NKCC1 inhibition by bumetanide. For this purpose, we combined electrophysiological and optical methods in slice preparations and assessed the properties of seizure like events (SLE) induced by 4-aminopyridine. In freeze-lesioned but not control slices, SLE onset was confined to the perilesional area, confirming that this region is hyperexcitable and likely triggers pathological activity. Bumetanide selectively reduced epileptic activity in lesion-containing slices but not in slices from sham-treated control rats. Moreover, bumetanide caused a shift in the SLE onset site away from the perilesional area. In contrast, effects of other antiepileptic drugs including carbamazepine, lacosamide, acezatolamide and zonisamide occurred mostly independently of the lesion and did not result in a shift of the onset region. Our work adds evidence for the functional relevance of chloride homeostasis in the pathophysiology of microgyrus formation as represented in the focal freeze-lesion model. Further studies in different MCD models and human tissue will be required to validate the effects across different MCD subtypes and species and to assess the translational value of our findings.
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Anticonvulsivantes/administración & dosificación , Criocirugía/efectos adversos , Malformaciones del Desarrollo Cortical/tratamiento farmacológico , Malformaciones del Desarrollo Cortical/patología , Convulsiones/tratamiento farmacológico , Convulsiones/patología , 4-Aminopiridina/toxicidad , Animales , Bumetanida/administración & dosificación , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/patología , Masculino , Malformaciones del Desarrollo Cortical/etiología , Técnicas de Cultivo de Órganos , Ratas , Ratas Wistar , Convulsiones/inducido químicamente , Inhibidores del Simportador de Cloruro Sódico y Cloruro Potásico/administración & dosificaciónRESUMEN
Up to date, preclinical screening for new antiepileptic substances is performed by a combination of different in vivo models of acute seizures, for which large numbers of animals are necessary. So far, little attention has been paid to in vitro models, which are also able to detect antiepileptic efficacy and in principle could likewise serve for exploratory preclinical screening. One of the established in vitro models of acute seizures is the 4-aminopyridine (4-AP) model. Previous studies have shown that the 4-AP model is capable to recapitulate the antiepileptic efficacy of standard antiepileptic drugs (AEDs) such as valproate or carbamazepine. Here, we employed a dual methodological approach using electrophysiology and optical imaging to systematically test the antiepileptic efficacy of three new-generation AEDs with distinct mechanisms of action (lacosamide, zonisamide, and levetiracetam). We found that frequency of 4-AP induced seizure like events (SLE) was the most sensitive parameter to detect dose-dependent antiepileptic effects in these compounds. Specifically, levetiracetam reduced SLE frequency while lacosamide and zonisamide at higher doses completely blocked SLE incidence. Analysis of the intrinsic optical signal additionally revealed a subiculum-specific reduction of the area involved in the propagation of ictal activity when lacosamide or zonisamide were administered. Taken together, our data adds some evidence that acute seizure models in vitro are in principle capable to detect antiepileptic effects across different mechanisms of action with efficacy similar to acute models in vivo. Further studies with negative controls, e.g., penicillin as a proconvulsant, and other clinically relevant AEDs are needed to determine if this acute in vitro model might be useful as exploratory screening tool. In view of the increasing sensitivity toward animal welfare, an affective in vitro model may help to reduce the number of laboratory animals deployed in burdening in vivo experiments and to preselect substances for subsequent testing in time- and cost-laborious models of chronic epilepsy.
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The dentate gyrus (DG) as part of the hippocampal formation is believed to serve as a gatekeeper with strong inhibitory properties against uncontrolled propagation of neuronal activity from the entorhinal cortex and neocortical structures. In temporal lobe epilepsy, the DG becomes hyperexcitable and loses its gate function, enabling propagation of ictal activity into downstream structures such as CA3 and CA1 areas. Furthermore, the DG, apart from facilitating propagation, may also be able to autonomously generate ictal activity, but this point has remained open so far. To tackle this question, we used intrinsic optical imaging in combination with electrophysiological recordings in brain slice preparations from rats in which status epilepticus had been induced electrically several weeks prior to measurements. Upon omission of Mg++ from the artificial cerebrospinal fluid, in 15 out of 33 slices (45.4%) from 9 out of 13 epileptic animals (69.2%), spontaneous and autonomous ictal activity, mostly seizure-like events (SLE), was observed in the DG. This activity manifested independently from SLE generated in adjacent cortices and never occurred in slices from control animals. SLE generated in the DG differed from those with origin in the entorhinal or temporal cortex by longer latency to the first event after Mg++ omission (p<0.001), a higher SLE frequency (p<0.05), higher amplitude (p<0.001) and a longer SLE duration (p<0.05). We conclude that in epilepsy, the DG, in addition to facilitated gating of activity from upstream structures, can serve as an independent generator of ictal activity.
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Giro Dentado/fisiopatología , Estado Epiléptico/fisiopatología , Animales , Modelos Animales de Enfermedad , Estimulación Eléctrica , Corteza Entorrinal/fisiopatología , Potenciales Postsinápticos Excitadores , Deficiencia de Magnesio , Masculino , Microelectrodos , Imagen Óptica , Ratas Wistar , Técnicas de Cultivo de TejidosRESUMEN
The subiculum is the principal target of CA1 pyramidal cells and mediates hippocampal output to various cortical and subcortical regions of the brain. The majority of subicular pyramidal cells are burst-spiking neurons. Previous studies indicated that high frequency stimulation in subicular burst-spiking cells causes presynaptic NMDA-receptor dependent long-term potentiation (LTP) whereas low frequency stimulation induces postsynaptic NMDA-receptor-dependent long-term depression (LTD). In the present study, we investigate the effect of 5-hydroxytryptamine type 4 (5-HT4) receptor activation and blockade on both forms of synaptic plasticity in burst-spiking cells. We demonstrate that neither activation nor block of 5-HT4 receptors modulate the induction or expression of LTP. In contrast, activation of 5-HT4 receptors facilitates expression of LTD, and block of the 5-HT4 receptor prevents induction of short-term depression and LTD. As 5-HT4 receptors are positively coupled to adenylate cyclase 1 (AC1), 5-HT4 receptors might modulate PKA activity through AC1. Since LTD is blocked in the presence of 5-HT4 receptor antagonists, our data are consistent with 5-HT4 receptor activation by ambient serotonin or intrinsically active 5-HT4 receptors. Our findings provide new insight into aminergic modulation of hippocampal output.
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Hipocampo/fisiología , Depresión Sináptica a Largo Plazo , Receptores de Serotonina 5-HT4/metabolismo , Sinapsis/fisiología , Animales , Potenciación a Largo Plazo , Masculino , Ratas , Ratas WistarRESUMEN
The subiculum (Sub) as a part of the hippocampal formation is thought to play a functional role in learning and memory. In addition to its major input from CA1 pyramidal cells, the subiculum receives input from the entorhinal cortex (EC) via the temporoammonic pathway. Thus far, synaptic plasticity in the subiculum was mainly investigated at CA1-Sub synapses. According to their spiking pattern, pyramidal cells in the subiculum were classified as bursting cells and non-bursting cells. In the present study, we demonstrate that subicular bursting cells show input-specific forms of long-term potentiation (LTP). At CA1-Sub synapses, bursting cells have been shown to express a presynaptic NMDA receptor-dependent LTP that depends on the activation of a cAMP-PKA cascade (Wozny et al., Journal of Physiology 2008). In contrast, at EC-Sub synapses the induction of LTP in bursting cells shows a high induction-threshold and relies on the activation of postsynaptic NMDA receptors, postsynaptic depolarization and postsynaptic Ca(2+) influx. Each form of LTP is input-specific and fails to induce heterosynaptic plasticity. Taken together, our data suggest that distinct, input-specific mechanisms govern high frequency-induced LTP at subicular bursting cells' synapses.