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LES ENREGISTREMENTS MULTI-ELECTRODE ARRAY (MEA): UNE VUE MACROSCOPIQUEMEA

Multi-Electrode Array
Figure 1A:A Multi-Electrode Array is an array made of microscopic metal electrodes (10-30 μm of diameter) distributed on a small surface.
Figure 1B:A Multi-Electrode Array is an array made of microscopic metal electrodes (10-30 μm of diameter) distributed on a small surface.

Pour quoi ?

La technologie du MEA permet de réaliser simultanément des enregistrements extracellulaires multi-sites sur une seule tranche de cerveau, fournissant ainsi une vue macroscopique exceptionnelle du fonctionnement des réseaux neuronaux.
Cette technique permet d’effectuer un grand nombre d’enregistrements dans des temps relativement courts (une à plusieurs semaines en fonction des tests) avec un bon rendement, pour le criblage et ou le profilage de composés par exemple.

Qu’est-ce qu’un MEA ?

Un Multi-Electrode Array est composé d’un réseau d’électrodes en métal microscopiques (électrodes de 10 à 30 μm de diamètre) distribuées sur une petite surface (~0.8-6 mm²). Elles peuvent être uniformément réparties ou agencées de façon à correspondre étroitement à l’organisation spatiale de la région cérébrale étudiée. Ces petites électrodes (recouvertes d’un métal inerte et biocompatible) sont utilisées pour enregistrer les signaux électriques liés à l’activité neuronale à l’intérieur d’une tranche de cerveau.

Quels sont les avantages des enregistrements avec les MEA ?

Votre composé est évalué in vitro… mais au plus proche des conditions physiologiques

Il est possible de préparer et garder en vie des tranches de cerveaux de rongeurs (250-500 μm d’épaisseur) pendant plusieurs heures consécutives. Ce type de tranches peut être préparé à partir de nombreuses régions différentes du cerveau et permet de garder intactes l’organisation et les connexions des cellules neuronales et gliales entre elles, une situation comparable à l’in vivo. De plus, récepteurs, canaux, enzymes sont à l’état natif et les voies de signalisation et de régulation cellulaires sont fonctionnelles.

Les enregistrements avec des MEA augmentent la robustesse des données

Le fait de pouvoir enregistrer en parallèle plusieurs réponses évoquées à l’aide de différentes électrodes, et ce à l’intérieur d’une seule et même tranche, donc dans la même région d’intérêt, augmente la fiabilité des données et leur traitement statistique ultérieur.

Les enregistrements de tranches de cerveau avec des MEA fournissent une vue macroscopique exceptionnelle des réseaux neuronaux

Des effets région-spécifiques peuvent être facilement observés grâce à la large surface couverte par les électrodes du MEA. A titre d’exemple, les activités spécifiques de composés sur certaines populations neuronales peuvent être documentées dans des structures laminaires telles que l’hippocampe ou le cortex.

Les enregistrements en MEA permettent le criblage en parallèle de plusieurs composés, avec rendement important

Les enregistrements réalisés à l’aide des MEA constituent une approche bien adaptée pour des tests fonctionnels en série : ils sont en effet beaucoup plus rapides que les enregistrements classiques (avec une électrode en verre) et ils peuvent être poussés à un haut niveau de standardisation.

PHARMACOLOGY: CHARACTERIZATION OF COMPOUND X EFFECT ON KAINATE-INDUCED GAMMA OSCILLATIONS RECORDED FROM RAT HIPPOCAMPAL SLICES
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SAFETY: EVALUATION OF 4 COMPOUNDS PRO-CONVULSIVE PROPERTIES IN A MODEL 4-AP-INDUCED EPILEPSY, IN RAT HIPPOCAMPAL SLICES
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LE PATCH-CLAMP: LA VUE MICROSCOPIQUEPatch-clamp

Pour quoi ?

Cette technologie offre la plus haute résolution pour les enregistrements électrophysiologiques à l’échelle unitaire du neurone (voir à l’échelle d’un seul canal ionique) grâce à différentes configurations d’enregistrements (cellule entière, patch perforé, cellule attachée, patch excisé « outside-out »). Des questions physiologiques ou pharmacologiques très précises peuvent être abordées avec cette technique, clairement dédiée à la compréhension des mécanismes d’action.

Comment ça marche ?

Les enregistrements en patch-clamp sont effectués à l’aide d’électrodes en verre (borosilicate) mesurant 1 à 2 μm de diamètre à leur extrémité.

La pipette, contenant une solution saline faite d’électrolytes, est hermétiquement scellée sur la membrane neuronale et en isole une partie (patch) électriquement. La pointe de la pipette permet un contact d’étanchéité d’un giga ohm avec la membrane. La plupart du temps le bout de membrane situé sous l’électrode est « aspiré » et rompu afin d’établir une totale continuité entre l’intérieur de la pipette et l’intérieur du neurone : on se trouve alors en configuration d’enregistrement « cellule entière ». Les flux d’ions traversant les canaux ioniques de la membrane neuronale sont alors enregistrés à l’aide d’un amplificateur différentiel très sensible. L’enregistrement de ce courant permet de tirer des conclusions sur le fonctionnement et la modulation des canaux ioniques insérés dans la membrane plasmique du neurone, voir de faire des analyses subrégionales (membrane pré ou post-synaptique).

Les avantages du Patch-Clamp

Les enregistrements en patch-clamp permettent :

Une évaluation in vitro de votre composé dans des conditions très proches des conditions physiologiques in vivo
Une étude approfondie de l’effet de votre composé à l’échelle moléculaire (récepteur, canal ionique)
De tester les effets de composés sur un large éventail de structures cérébrales, sur différents types cellulaires ou sur certains types de neurotransmetteurs et/ou récepteurs
D’obtenir des données de haute qualité : robustes et reproductibles
CNS: EFFECT OF A7 NICOTINIC RECEPTOR AGONIST ON GABAERGIC SYNAPTIC ACTIVITY IN HIPPOCAMPUS
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PAIN: PERIPHERAL PAIN AND NICOTINIC RECEPTOR MODULATION
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CULTURE CELL: EFFECT OF A POTENTIAL ALLOSTERIC MODULATOR OF NR2B-CONTAINING NMDA RECEPTORS ON HIPPOCAMPAL CULTURE
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HUMAN BRAIN SLICES RECORDINGS
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Acetylcholinesterase+
Acetylcholine receptors STUDIES
Acetylcholinesterase
Evaluation of 3 compounds on short-term plasticity in the rat hippocampal slice Evaluation of 2 compounds on LTP recorded in the CA1 region of the adult rat hippocampal slice (2)
M3 receptors+
Acetylcholine receptors STUDIES
M3 receptors
Evaluation of 3 compounds on CA1 neurons spontaneous firing (effect normalized to 1 µM Carbachol-induced firing)
M1 receptors+
Acetylcholine receptors STUDIES
M1 receptors
Evaluation of 3 compounds on CA1 neurons spontaneous firing (effect normalized to 1 µM Carbachol-induced firing) (10 studies) Evaluation of 1 compound on CA1 neurons spontaneous firing in the presence and in the absence of Pirenzepine (1) Determination of one compound dose-effect on synaptic transmission in the CA1 region of rat hippocampal slices, alone and in the presence of a low dose of Oxotremorine
α7 nicotinic receptors+
Acetylcholine receptors STUDIES
α7 nicotinic receptors
Peripheral pain and nicotinic receptor modulationStudy of PAMS (Positive Allosteric Modulator) at α7 nicotinic receptors expressed by spinal cord neurons.Effects of α7 positive allosteric modulator (PAM) on ACh-evoked responses in juvenile rat spinal cord (layers II/III)Effects of α7 positive allosteric modulator (PAM) on ACh-evoked responses in juvenile rat spinal cord laminal/IIStudy of PAMS (Positive Allosteric Modulator) at α7 nicotinic receptors expressed by spinal cord neurons Evaluation of α7 nicotinic receptor agonists on GABAergic synaptic activity in hippocampus
α4 nicotinic receptors+
Acetylcholine receptors STUDIES
α4 nicotinic receptors
Evaluation of compounds on Short- and Long-Term Potentiation in CA1 and DG region of rat hippocampal slice
Ca channels+
Calcium channels STUDIES
Ca channels
Evaluation of 3 compounds (step dosing + TTX) on Basal Synaptic Transmission in the CA1 region of mice hippocampal slices
A2 receptors+
Adenosine receptors STUDIES
A2 receptors
Evaluation of 1 reference and 1 assumed A2A receptors agonist on short- and long-term plasticity properties in the CA1 region of the hippocampal slice
Slo 2.2+
Potassium channels STUDIES
Slo 2.2
Patch-clamp study for Slo 2.2 characterization in WT & Tg mice
Kv7+
Potassium channels STUDIES
Kv7
Evaluation of Compound X effect on Low-magnesium-induced ED in rat cortical and hippocampal slices Evaluation of 2 compounds at 1, 3 and 10 µM on low-magnesium-induced ED in rat hippocampal slices
K channels+
Potassium channels STUDIES
K channels
Evaluation of compounds on Paired-pulse protocols in the CA1 region of adult rat hippocampus
Noradrenaline transporters+
Adrenergic receptors STUDIES
Noradrenaline transporters
Oxygen-Glucose Deprivation (OGD) assay development
α receptors+
Adrenergic receptors STUDIES
α receptors
Physiological and Pharmacological Profiling of Compound X on the Adult Rat Hippocampal Slice based on Electrophysiological Recordings Physiological and pharmacological profiling of low Compound X concentration on the adult rat hippocampal slice based on electrophysiological recordings Investigation of a dose-response effect of Compound X at LPP-DG synapse in the rat hippocampus
Na Channels+
Sodium channels STUDIES
Na Channels
Evaluation of 1 compound on spontaneous firing of neurons (CA1 + PN)
Dopamine transporters+
Dopamine Receptors STUDIES
Dopamine transporters
Evaluation of one compound effect on weak tetanus-induced potentiation (2)
D1/D5 receptors+
Dopamine Receptors STUDIES
D1/D5 receptors
dopaminergic receptors+
Dopamine Receptors STUDIES
dopaminergic receptors
Physiological and Pharmacological Profiling of Compound X on the Adult Rat Hippocampal Slice based on Electrophysiological Recordings Physiological and pharmacological profiling of low Compound X concentration on the adult rat hippocampal slice based on electrophysiological recordings Investigation of a dose-response effect of Compound X at LPP-DG synapse in the rat hippocampus
Studies+
Phenotyping STUDIES
Studies
Phenotyping and evaluation of one compound on synaptic transmission and plasticity in WT and KO miceInvestigation of basal synaptic transmission, short-term plasticity and Long-Term Depression in hippocampal slices from WT and Transgenic mice Characterisation of Long-Term Potentiation profiles in a transgenic mouse model and its wild-type littermateCharacterisation of Long-Term Potentiation profiles in two transgenic mice strains and comparison with wild-type data Characterization of LTP profiles in WT & KO mice Characterization of LTP profiles in WT versus 2 KO strains (mice)Investigation of short-and long-term plasticity properties in hippocampal slices from WT and KO mice
GABAB receptors+
GABA Receptors STUDIES
GABAB receptors
Evaluation of one compound at 3 concentrations on Epileptiform Discharges induced by a zero-magnesium ACSF in rat hippocampal slices
GABAA receptors +
GABA Receptors STUDIES
GABAA receptors
Evaluation of diazepam and one compound on miniature IPSCs and evoked EPSCs in mouse spinal cord Effect of one compound on the inhibitory synaptic activity in the CA1 region Evaluation of pro-convulsive effect of 4 molecules and comparison to bicuculline and picrotoxin in adult rat brain slices Evaluation of Compound X on GABAergic-mediated inhibition in CA1 region of rat hippocampal slice Evaluation of Compound X properties towards NMDA-induced excitotoxicity in rat hippocampal slices Evaluation of one compound on GABAergic-mediated inhibition in CA1 region of the hippocampal slice using Multi-Electrode Array (PPI protocol) Evaluation of 2 compounds on GABAercic-mediated inhibition in CA1 region of the hippocampal slice (PPI protocol)
GABA receptors+
GABA Receptors STUDIES
GABA receptors
Evaluation of compounds on Paired-pulse protocols in the CA1 region of adult rat hippocampus
mGlu4 receptors+
Glutamate receptors STUDIES
mGlu4 receptors
POC : Electrophysiological characterisation of two Positive Allosteric Modulators (PAM) of metabotropic GluR4 receptors Electrophysiological characterization of Compound X effect at LPP synapsesEvaluation of 2 assumed mGluR4 PAM at LPP synapses (3)
mGlu2/3 receptors+
Glutamate receptors STUDIES
mGlu2/3 receptors
Evaluation of 2 compounds on DCG-IV mediated-inhibition of synaptic transmission at MPP synapses Evaluation of two compounds on paired-pulse depression and synaptic transmission at MPP synapses in rat hippocampal slices Evaluation of Pharmacological properties of Compound X (a Positive Allosteric Modulator of mGluR2 Receptors) at the MPP-Granule Cells Synapse Evaluation of Pharmacological properties of 2 compounds (Positive Allosteric Modulators of mGluR2 Receptors) at the MPP-Granule Cells Synapse Characterisation of 3 compounds on synaptic transmission and plasticity at MPP-DG synapses in the adult rat hippocampal slice Evaluation of Compound X on DCG-IV- induced inhibition of synaptic transmission at MPP-DG synapses in the rat hippocampal slice
mGlu1/5 receptors+
Glutamate receptors STUDIES
mGlu1/5 receptors
Evaluation of Compound X effect on DHPG-induced LTD in the CA1 region of rat hippocampal slices (2)
mGlu5 receptors+
Glutamate receptors STUDIES
mGlu5 receptors
Evaluation of five compounds on 100 Hz induced LTP in the CA1 region of adult rat hippocampal slice
AMPA/Kainate receptors+
Glutamate receptors STUDIES
AMPA/Kainate receptors
Evaluation of 4 compounds effect on AMPA/KA- and NMDA-mediated EPSP in rat hippocampal slices
Kainate receptors+
Glutamate receptors STUDIES
Kainate receptors
Evaluation of Bacopa effects on weak tetanus-induced potentiation in hippocampal slices
AMPA receptors+
Glutamate receptors STUDIES
AMPA receptors
Evaluation of PS-DHA effects on weak tetanus-induced potentiation in hippocampal slices
NMDA receptors+
Glutamate receptors STUDIES
NMDA receptors
Effect of a compound treatment on passive and active membrane properties of medium spiny neurons Determination of the effects of compounds on Long-Term Potentiation in Hippocampus Evaluation of compounds effects on weak tetanus-induced potentiation in hippocampal slices Evaluation of compounds effects on weak tetanus-induced potentiation in hippocampal slices Determination of the effects of 2 compounds on weak tetanus-induced potentiation in the hippocampus Evaluation of 2 compounds, alone and in combination on weak tetanus-induced potentiation in hippocampal slices Evaluation of the effect of compounds in combination on weak tetanus-induced potentiation in mice hippocampal slices Evaluation of compound effects on weak tetanus-induced potentiation in hippocampal slices Evaluation of compound effects on weak tetanus-induced potentiation in mice hippocampal slices Evaluation of one compound acute effect on rat hippocampal slices Evaluation of 4 compounds effect on AMPA/KA- and NMDA-mediated EPSP in rat hippocampal slices Investigation of 2 compounds effect on NMDA-mediated neurotransmission and CA1 neurons firing in rat hippocampal slices Evaluation of one compound on Short-Term Potentitiation (STP) Evaluation of one compound on purely NMDA-mediated EPSP Evaluation of 2 compounds on MK-801-impaired LTP in CA1 region of the hippocampal slice Evaluation of compounds on NMDA receptors
H3 receptors+
Histamine receptors STUDIES
H3 receptors
Comparison of five H3-histamine receptors antagonist/inverse agonist properties in the adult rat hippocampal sliceCharacterization of one neutral antagonist and two inverse agonist of H3-histamine receptors properties in adult rat hippocampal slice
H1, H2, H3 receptors+
Histamine receptors STUDIES
H1, H2, H3 receptors
Effect of Compound X on histaminergic receptors (sub-types H1, H2 and H3) assessed with electrophysiological recordings of hippocampal slices of adult rats (Sprague-Dawley) on 3D multi-electrode arrays
Melatonin receptors+
Melatonin receptors STUDIES
Melatonin receptors
Electrophysiological characterization of Melatonin and Agomelatine effect son Short- and Long-Term Potentiation and CA1 neurons firing in mice and rat hippocampal slices
ORL receptors+
Opiod receptors STUDIES
ORL receptors
Evaluation of three compounds on TBS-induced LTP in the CA1 region of the adult mouse hippocampus Establish pharmacologically relevant concentrations of the Compound X in this in vitro slice assay by inhibition of Nociceptin-induced effect on fEPSPs (2)
δ receptors+
Opiod receptors STUDIES
δ receptors
Investigation of pro-convulsive properties of Compound X Investigation of the pharmacological profile of Compound X in the CA1 region of the adult rat hippocampal sliceCharacterization of seven compounds together with Compound X in the CA1 region of the adult rat hippocampal slice (2)
NPY receptors+
Peptid receptors STUDIES
NPY receptors
Evaluation of 1 compound on Y5 receptor mediated inhibition at MF synapses Evaluation of compounds on LTP in adult rat hippocampal slice
5-HT6+
Serotoninergic receptors STUDIES
5-HT6
Evaluation of 4 compounds on PPI and LTP in CA1 region of the hippocampal slice (recording inside SP/SR)
5-HT2B+
Serotoninergic receptors STUDIES
5-HT2B
Effect of Metadoxine on medium spiny neuronsEffect of metadoxine on mEPSC frequency and amplitude
5-HT6/7+
Serotoninergic receptors STUDIES
5-HT6/7
Physiological and Pharmacological Profiling of Compound X on the Adult Rat Hippocampal Slice based on Electrophysiological Recordings Physiological and pharmacological profiling of low Compound X concentration on the adult rat hippocampal slice based on electrophysiological recordings Investigation of a dose-response effect of Compound X at LPP-DG synapse in the rat hippocampus
5-HT transporter+
Serotoninergic receptors STUDIES
5-HT transporter
Evaluation of 3 compounds on Long-Term Potentiation in the CA1 region of rat hippocampal slice
5-HT+
Serotoninergic receptors STUDIES
5-HT
Characterization of Compound X effect on Kainate-induced oscillations recorded from rat hippocampal slicesEvaluation of Compound X effects on synaptic transmission and plasticity between CA3 and CA1 region of adult rat hippocampal slicesEvaluation of 3 compounds on Long-Term Potentiation in the CA1 region of rat hippocampal slice Evaluation of 4 compounds on PPI and LTP in CA1 region of the hippocampal slice (recording inside SP/SR)
GlyR+
Glycine receptors STUDIES
GlyR
Effect of compounds on glycine-evoked currents
TAAR1+
G protein-coupled receptor STUDIES
TAAR1
Characterization in the VTA DOPA, GABA neuron comparison of the spontaneous firing frequency and evaluation of a compound and the potential block of the effect by another compound in WT and TAAR1-OE miceEvaluation of a compound on the firing rate in mice acute dorsal raphe slicesEvaluation of a compound on the firing rate in acute dorsal raphe slices of miceCharacterization of TAAR1 in acute dorsal raphe slices of miceEvaluation of a compound on the firing rate in acute slices of the ventral tegmental areaEvaluation of a compound on the firing rate in acute ventral tegmental area slices of miceCharacterization of some TAAR1-mediated effects in acute striatal slices of mice

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