The Metabolic Disturbances of Motoneurons Exposed to Glutamate
Glutamate-induced excitotoxicity is considered as one of the major pathophysiological factors of motoneuron death in amyotrophic lateral sclerosis and other motoneuron diseases. In order to expand our knowledge on mechanisms of glutamate-induced excitotoxicity, the present study proposes to determine the metabolic consequences of glutamate and astrocytes in primary enriched motoneuron culture. Using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS), we showed that the presence of astrocytes and glutamate profoundly modified the metabolic profile of motoneurons. Our study highlights for the first time that crosstalk between astrocytes and enriched motoneuron culture induced alterations in phenylalanine, tryptophan, purine, arginine, proline, aspartate, and glutamate metabolism in motoneurons. We observed that astrocytes modulate the sensitivity of motoneurons to glutamate, since metabolites altered by glutamate in motoneurons cultured alone were different (except 5-hydroxylysine) from those altered in co-cultured motoneurons. Our findings provide new insight into the metabolic alterations associated to astrocytes and glutamate in motoneurons and provide opportunities to identify novel therapeutic targets.
KeywordsFingerprinting Excitotoxicity Metabolomics Amyotrophic lateral sclerosis ALS Primary motoneuron cultures Astrocytes Metabolites
We thank the staff of the Programme Pluri-Formation “Analyze des Systèmes Biologiques” (PPF ASB) platform of the University François-Rabelais in Tours, France, Céline Salsac, and Antoine Lefèvre for technical assistance.
B. MH, H. B, E. C, P. V, P. E, P. C, C. R. A, C. R, and S. M designed of the experiments and wrote the manuscript. H. B and S. M managed the project. B. MH, E. C, C. R, and S. M performed experiments and/or contributed to the interpretation of data.
This work was supported by grants from the institut national de la santé et de la recherche médicale (INSERM), the association française pour la recherche sur la SLA (ARSLA), ANR-14-RARE-0006 E-RARE “FasSMALS” and ANR “GliALS”. E.C. received a grant from the association française contre les myopathies (AFM) and B. MH received a grant from “La Région Centre” (2013–10).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- 6.Madji Hounoum B, Vourc'h P, Felix R, Corcia P, Patin F, Gueguinou M, Potier-Cartereau M, Vandier C et al (2016) NSC-34 motor neuron-like cells are unsuitable as experimental model for glutamate-mediated excitotoxicity. Front Cell Neurosci 10:118. https://doi.org/10.3389/fncel.2016.00118 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Madji Hounoum B, Mavel S, Coque E, Patin F, Vourc'h P, Marouillat S, Nadal-Desbarats L, Emond P et al (2017) Wildtype motoneurons, ALS-linked SOD1 mutation and glutamate profoundly modify astrocyte metabolism and lactate shuttling. Glia 65(4):592–605. https://doi.org/10.1002/glia.23114 CrossRefPubMedGoogle Scholar
- 13.Raoul C, Estevez AG, Nishimune H, Cleveland DW, deLapeyriere O, Henderson CE, Haase G, Pettmann B (2002) Motoneuron death triggered by a specific pathway downstream of Fas. Potentiation by ALS-linked SOD1 mutations. Neuron 35(6):1067–1083. https://doi.org/10.1016/S0896-6273(02)00905-4 CrossRefPubMedGoogle Scholar
- 15.Zhang H, Xing L, Rossoll W, Wichterle H, Singer RH, Bassell GJ (2006) Multiprotein complexes of the survival of motor neuron protein SMN with Gemins traffic to neuronal processes and growth cones of motor neurons. J Neurosci 26(33):8622–8632. https://doi.org/10.1523/jneurosci.3967-05.2006 CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Madji Hounoum B, Blasco H, Nadal-Desbarats L, Diémé B, Montigny F, Andres CR, Emond P, Mavel S (2015) Analytical methodology for metabolomics study of adherent mammalian cells using NMR, GC-MS and LC-HRMS. Anal Bioanal Chem 407(29):8861–8872. https://doi.org/10.1007/s00216-015-9047-x CrossRefPubMedGoogle Scholar
- 24.Rothstein JD, Dykes-Hoberg M, Pardo CA, Bristol LA, Jin L, Kuncl RW, Kanai Y, Hediger MA et al (1996) Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate. Neuron 16(3):675–686. https://doi.org/10.1016/S0896-6273(00)80086-0 CrossRefPubMedGoogle Scholar
- 27.Daniels BP, Jujjavarapu H, Durrant DM, Williams JL, Green RR, White JP, Lazear HM, Gale M Jr et al (2017) Regional astrocyte IFN signaling restricts pathogenesis during neurotropic viral infection. J Clin Invest 127(3):843–856. https://doi.org/10.1172/JCI88720 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Baslow MH (2010) Evidence that the tri-cellular metabolism of N-acetylaspartate functions as the brain's "operating system": How NAA metabolism supports meaningful intercellular frequency-encoded communications. Amino Acids 39(5):1139–1145. https://doi.org/10.1007/s00726-010-0656-6 CrossRefPubMedGoogle Scholar
- 31.Gerardo-Nava J, Mayorenko II, Grehl T, Steinbusch HW, Weis J, Brook GA (2013) Differential pattern of neuroprotection in lumbar, cervical and thoracic spinal cord segments in an organotypic rat model of glutamate-induced excitotoxicity. J Chem Neuroanat 53:11–17. https://doi.org/10.1016/j.jchemneu.2013.09.007 CrossRefPubMedGoogle Scholar
- 32.Do-Ha D, Buskila Y, Ooi L (2017) Impairments in motor neurons, interneurons and astrocytes contribute to Hyperexcitability in ALS: Underlying mechanisms and paths to therapy. Mol Neurobiol. https://doi.org/10.1007/s12035-017-0392-y
- 35.Pellerin L, Magistretti PJ (2005) Ampakine CX546 bolsters energetic response of astrocytes: A novel target for cognitive-enhancing drugs acting as alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor modulators. J Neurochem 92(3):668–677. https://doi.org/10.1111/j.1471-4159.2004.02905.x CrossRefPubMedGoogle Scholar
- 38.Arumugam S, Garcera A, Soler RM, Tabares L (2017) Smn-deficiency increases the intrinsic excitability of motoneurons. Front Cell Neurosci 11(269). https://doi.org/10.3389/fncel.2017.00269
- 39.Urushitani M, Shimohama S, Kihara T, Sawada H, Akaike A, Ibi M, Inoue R, Kitamura Y et al (1998) Mechanism of selective motor neuronal death after exposure of spinal cord to glutamate: Involvement of glutamate-induced nitric oxide in motor neuron toxicity and nonmotor neuron protection. Ann Neurol 44(5):796–807. https://doi.org/10.1002/ana.410440514 CrossRefPubMedGoogle Scholar