Real-Time Monitoring of Levetiracetam Effect on the Electrophysiology of an Heterogenous Human iPSC-Derived Neuronal Cell Culture Using Microelectrode Array Technology.

Andrea Di Credico,Angela Di Baldassarre,Giulia Gaggi,Pascal Izzicupo,Giovanni Iannetti,Laura Ferri,Laura Bonanni,Barbara Ghinassi

doi:10.3390/bios11110450

Andrea Di Credico, Angela Di Baldassarre + Show 6 more

Open Access

https://doi.org/10.3390/bios11110450

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Abstract

Levetiracetam (LEV) is a broad-spectrum and widely used antiepileptic drug that also has neuroprotective effects in different neurological conditions. Given its complex interaction with neuronal physiology, a better comprehension of LEV effects on neurons activity is needed. Microelectrode arrays (MEAs) represent an advanced technology for the non-invasive study of electrophysiological activity of neuronal cell cultures. In this study, we exploited the Maestro Edge MEA system, a platform that allows a deep analysis of the electrical network behavior, to study the electrophysiological effect of LEV on a mixed population of human neurons (glutamatergic, GABAergic and dopaminergic neurons, and astrocytes). We found that LEV significantly affected different variables such as spiking, single-electrode bursting, and network bursting activity, with a pronounced effect after 15 min. Moreover, neuronal cell culture completely rescued its baseline activity after 24 h without LEV. In summary, MEA technology confirmed its high sensitivity in detecting drug-induced electrophysiological modifications. Moreover, our results allow one to extend the knowledge on the electrophysiological effects of LEV on the complex neuronal population that resembles the human cortex.

Highlights

Levetiracetam (LEV) is a second-generation, broad-spectrum antiepileptic, generally prescribed for the treatment of myoclonic, partial, and primary generalized tonic-clonic seizures [1,2]
Little is known about the mechanisms through which LEV exerts its anti-epileptic action [3]; they seem to include the binding to the synaptic vesicle protein 2A
To provide an in-depth report of the neuronal activity both during and after the treatment with LEV, we analyzed the spiking, which describes the number and regularity of action potentials occurrence; the single-electrode bursting, which defines the occurrence of bursts; the network bursting which encompasses several variables related to the network bursts, which are coordinated clusters of spiking across multiple tivities, and the evaluation of multiple extracellular spikes amplitude were performed

Summary

Introduction

Levetiracetam (LEV) is a second-generation, broad-spectrum antiepileptic, generally prescribed for the treatment of myoclonic, partial, and primary generalized tonic-clonic seizures [1,2]. MEAs consist of a high number of electrodes that can be of different materials (e.g., gold, poly(3,4ethylenedioxythiophene) polystyrene sulfonate—PEDOT) [9] usually implanted at the base of tissue culture wells that allows to record (and eventually stimulate) the activity of cells capable to generate electrical signals (e.g., neurons, cardiomyocytes) [10,11,12] in a label-free manner and in real-time. MEAs allow one to record the Extracellular Action Potentials (EAP, called spikes) [10,13]. In addition to the spike rate and timing, last-generation MEAs can provide higher-level analysis, recording singleelectrode bursting and network bursting activity [14]. These features provide important information regarding the electrophysiological activity and connectivity inside the neuronal cell culture [15]. EAP waveform analysis is an interesting tool to consider since it was seen that pharmacological treatment can alter its amplitude and waveform [8,18], and different neuron types can produce different waveforms [16]

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