title: Chronic mitochondrial inhibition induces glutamate-mediated corticomotoneuron death in an organotypic culture model.
abstract: There is growing evidence that mitochondrial dysfunction is an important factor in a cascade of neurotoxic events as observed during pathogenesis of various neurodegenerative diseases. In the neurodegenerative disease amyotrophic lateral sclerosis (ALS) both spinal and cortical motoneurons degenerate, but in experimental studies most attention so far has been focussed on the spinal motoneurons. In order to study the role of mitochondrial dysfunction in the pathways leading to cortical (upper) motoneuron (CMN) death, a long-term culture system of rat cortical explants was used. CMNs were visualized by immunocytochemical labeling with antibodies directed against nonphosphorylated neurofilament, SMI-32, and for their identification we also used their location in layer V of the explant, their size, and their morphological appearance. In this model the effect of mitochondrial inhibition was studied through chronic malonate treatment. For 2 weeks, low doses of complex II inhibitor malonate were added to the cultures twice a week. The malonate-induced chronic mitochondrial inhibition resulted in a dose-dependent increase of CMN death in the slices. Neuroprotection was achieved with the NMDA antagonist MK-801 and the non-NMDA antagonist CNQX indicating the involvement of glutamate in the malonate-induced CMN death. Furthermore, our data indicate that chronic mitochondrial inhibition results in CMN death, which is mediated by glutamate excitotoxicity via both non-NMDA and NMDA receptors. In this respect the present in vitro approach may act as a model for understanding mechanisms underlying CMN death in ALS.
authors.affiliation: Department of Experimental Neurology, RMI for Neurosciences, UMC Utrecht, 3508 GA, Utrecht, The Netherlands. m.g.h.vanwesterlaak@neuro.azu.nl
authors.forename: M G
authors.initials: MG
authors.lastname: Van Westerlaak
authors.forename: E A
authors.initials: EA
authors.lastname: Joosten
authors.forename: A A
authors.initials: AA
authors.lastname: Gribnau
authors.forename: A R
authors.initials: AR
authors.lastname: Cools
authors.forename: P R
authors.initials: PR
authors.lastname: Bär
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D048429:Cell Size; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018698:Glutamic Acid / Q000378:metabolism* / Q000633:toxicity; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008314:Malonates / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D008954:Models, Biological; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019636:Neurodegenerative Diseases / Q000209:etiology / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology; D017966:Pyramidal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
chemical_list: D000943:Antigens, Differentiation; D018691:Excitatory Amino Acid Antagonists; D008314:Malonates; D016900:Neurofilament Proteins; D018696:Neuroprotective Agents; D018698:Glutamic Acid; D016291:Dizocilpine Maleate; C030290:malonic acid