Chronic Electrical Stimulation Promotes the Excitability and Plasticity of ESC-derived Neurons following Glutamate-induced InhibitionIn vitro

Abstract

Functional electrical stimulation (FES) is rapidly gaining traction as a therapeutic tool for mediating the repair and recovery of the injured central nervous system (CNS). However, the underlying mechanisms and impact of these stimulation paradigms at a molecular, cellular and network level remain largely unknown. In this study, we used embryonic stem cell (ESC)-derived neuron and glial cocultures to investigate network maturation following acute administration of Lglutamate, which is a known mediator of excitotoxicity following CNS injury. We then modulated network maturation using chronic low frequency stimulation (LFS) and direct current stimulation (DCS) protocols. We demonstrated that L-glutamate impaired the rate of maturation of ESC-derived neurons and glia immediately and over a week following acute treatment. The administration of chronic LFS and DCS protocols individually following L-glutamate infusion significantly promoted the excitability of neurons as well as network synchrony, while the combination of LFS/DCS did not. qRT-PCR analysis revealed that LFS and DCS alone significantly up-regulated the expression of excitability and plasticity-related transcripts encoding N-methyl-D-aspartate (NMDA) receptor subunit (NR2A), brain-derived neurotrophic factor (BDNF) and Ras-related protein (RAB3A). In contrast, the simultaneous administration of LFS/DCS down-regulated BDNF and RAB3A expression. Our results demonstrate that LFS and DCS stimulation can modulate network maturation excitability and synchrony following the acute administration of an inhibitory dose of L-glutamate, as well as an upregulation of NR2A, BDNF and RAB3A gene expression. Our study also provides a novel framework for investigating the effects of electrical stimulation on neuronal responses and network formation/repair after traumatic brain injury.

Direct Current Stimulation in Cell Culture Systems and Brain Slices—New Approaches for Mechanistic Evaluation of Neuronal Plasticity and Neuromodulation: State of the Art

“…The intervention enhanced neuronal excitability as well as network synchronization. They furthermore demonstrated an upregulation of the NMDA receptor subunit NR2A, and Ras-related protein RAB3A in mouse Hb9 ESC-derived neurons and glial cells [ 50 ].…”

A guide to the manufacture of sustainable, ready to use in vitro platforms for the electric-field pacing of cellularised 3D porous scaffolds

“…In this work, we detail methods to rapid prototype an electrical pacing bioreactor and R3S -a Rig for Stimulation of Sponge-like Scaffolds. As a proof of concept and validation we demonstrate that these systems are compatible with isotropic and anisotropic porous scaffolds composed of collagen or poly (3,4-ethylene dioxythiophene):polystyrene sulfonate (PEDOT:PSS). External pacing of C3H10 cells on anisotropic porous scaffolds led to a metabolic increase and enhanced cell alignment.…”

“…Such stimuli include strain and stress, pressure, flow, and the subject focus of this paper: electrical pacing. Pacing is a method commonly used with engineered heart tissues derived from primary cardiomyocytes and cardiomyocytes derived from pluripotent stem cell sources 2 , neural applications in the case of neuronal cultures 3 , and skeletal muscle cultures 4 . More recently, the use of electrical pacing has been explored in non-traditional tissues such as skin 5 , bone 6 , and immune cells 7 .…”

The effects of electrical stimulation on glial cell behaviour

“…Direct current is often used in applications such as transcranial direct current stimulation (tDCS), which (in the clinical context) makes use of electrodes placed on the outside of the head and is designed to treat disorders such as depression and Parkinson's disease. Latchoumane et al investigated the molecular pathways underlying the treatment effects of tDCS [54]. Embryonic stem cell-derived neuron-glia co-cultures were subjected to chronic low frequency stimulation and direct current stimulation paradigms in the presence of the excitotoxic mediator L-glutamate to simulate CNS injury.…”