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The Journal of Neuroscience, August 27, 2003, 23(21):7881-7888
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Glutamate Decreases Mitochondrial Size and Movement in Primary Forebrain Neurons
Gordon L. Rintoul, Anthony J. Filiano, Jacques B. Brocard, Geraldine J. Kress, andIan J. Reynolds Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261
Mitochondria are essential to maintain neuronal viability. In addition to the generation of ATP and maintenance of calcium homeostasis, the effective delivery of mitochondria to the appropriate location within neurons is also likely to influence their function. In this study we examined mitochondrial movement and morphology in primary cultures of rat forebrain using a mitochondrially targeted enhanced yellow fluorescent protein (mt-eYFP). Mt-eYFP-labeled mitochondria display a characteristic elongated phenotype and also move extensively. Application of glutamate to cultures results in a rapid diminution of movement and also an alteration from elongated to rounded morphology. This effect required the entry of calcium and was mediated by activation of the NMDA subtype of glutamate receptor. Treatment of cultures with an uncoupler or blocking ATP synthesis with oligomycin also stopped movement but did not alter morphology. Interestingly, application of glutamate together with the uncoupler did not prevent the changes in movement or shape but facilitated recovery after washout of the stimuli. This suggests that the critical target for calcium in this paradigm is cytosolic. These studies demonstrate that in addition to altering the bioenergetic properties of mitochondria, neurotoxins can also alter mitochondrial movement and morphology. We speculate that neurotoxin-mediated impairment of mitochondrial delivery may contribute to the injurious effects of neurotoxins.
Key words: green fluorescent protein; cytoskeleton; NMDA receptor; intracellular calcium; excitotoxicity; organelle transport
Received Feb 24, 2003; revised June 23, 2003; accepted July 2, 2003.
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