Abstract
Glutamate-induced excitotoxicity is a major contributor to motor neuron degeneration in the pathogenesis of amyotrophic lateral sclerosis (ALS). The spinal cord × Neuroblastoma hybrid cell line (NSC-34) is often used as a bona fide cellular model to investigate the physiopathological mechanisms of ALS. However, the physiological response of NSC-34 to glutamate remains insufficiently described. In this study, we evaluated the relevance of differentiated NSC-34 (NSC-34D) as an in vitro model for glutamate excitotoxicity studies. NSC-34D showed morphological and physiological properties of motor neuron-like cells and expressed glutamate receptor subunits GluA1–4, GluN1 and GluN2A/D. Despite these diverse characteristics, no specific effect of glutamate was observed on cultured NSC-34D survival and morphology, in contrast to what has been described in primary culture of motor neurons (MN). Moreover, a small non sustained increase in the concentration of intracellular calcium was observed in NSC-34D after exposure to glutamate compared to primary MN. Our findings, together with the inability to obtain cultures containing only differentiated cells, suggest that the motor neuron-like NSC-34 cell line is not a suitable in vitro model to study glutamate-induced excitotoxicity. We suggest that the use of primary cultures of MN is more suitable than NSC-34 cell line to explore the pathogenesis of glutamate-mediated excitotoxicity at the cellular level in ALS and other motor neuron diseases.
Highlights
Amyotrophic lateral sclerosis (ALS) is one of the most common neurodegenerative diseases in adults, caused by the selective death of motor neurons (MN)
When Neuroblastoma × Spinal Cord (NSC)-34 cells were cultured in the differentiation media (NSC-34D), we distinguished two morphologically distinct populations: cells with short neurites, and cells with phenotypic characterization of MN with long processes (Figure 1A)
We investigated whether the potent morphogen retinoic acid (RA) could enhance differentiation of NSC-34 into motor neuron-like cells
Summary
Amyotrophic lateral sclerosis (ALS) is one of the most common neurodegenerative diseases in adults, caused by the selective death of motor neurons (MN). Glutamate excitotoxicity is a major contributor to dysfunction and death of MN in the pathogenesis of ALS (Heath and Shaw, 2002; Van Den Bosch et al, 2006; Spalloni et al, 2013; Blasco et al, 2014). Overstimulation of glutamate receptors facilitates the entry and the excess of calcium (Ca2+) in cell compartments, leading to a cascade of destructive events by calcium-dependent enzymatic pathways and mitochondrial dysfunction with the generation of free radicals (Van Den Bosch et al, 2000; Blasco et al, 2014). AMPA and NMDA receptors are largely responsible for calcium flux across neuronal cell membranes (Hollmann et al, 1991; Van Den Bosch et al, 2000)
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