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dc.contributor.authorUgbode, Christopher I.*
dc.contributor.authorHirst, W.D.*
dc.contributor.authorRattray, Marcus*
dc.date.accessioned2016-02-23T16:40:36Z
dc.date.available2016-02-23T16:40:36Z
dc.date.issued2014-09
dc.identifier.citationUgbode C, Hirst WD and Rattray M (2014) Neuronal influences are necessary to produce mitochondrial co-localization with glutamate transporters in astrocytes. Journal of Neurochemistry, 130 (5): 668-77.en_US
dc.identifier.urihttp://hdl.handle.net/10454/7811
dc.descriptionyesen_US
dc.description.abstractAbstract Recent evidence suggests that the predominant astrocyte glutamate transporter, GLT-1/ Excitatory Amino Acid Transporter 2 (EAAT2) is associated with mitochondria. We used primary cultures of mouse astrocytes to assess co-localization of GLT-1 with mitochondria, and tested whether the interaction was dependent on neurons, actin polymerization or the kinesin adaptor, TRAK2. Mouse primary astrocytes were transfected with constructs expressing V5-tagged GLT-1, pDsRed1-Mito with and without dominant negative TRAK2. Astrocytes were visualized using confocal microscopy and co-localization was quantified using Volocity software. Image analysis of confocal z-stacks revealed no co-localization between mitochondria and GLT-1 in pure astrocyte cultures. Co-culture of astrocytes with primary mouse cortical neurons revealed more mitochondria in processes and a positive correlation between mitochondria and GLT-1. This co-localization was not further enhanced after neuronal depolarization induced by 1 h treatment with 15 mM K+. In pure astrocytes, a rho kinase inhibitor, Y27632 caused the distribution of mitochondria to astrocyte processes without enhancing GLT-1/mitochondrial co-localization, however, in co-cultures, Y27632 abolished mitochondrial: GLT-1 co-localization. Disrupting potential mitochondrial: kinesin interactions using dominant negative TRAK2 did not alter GLT-1 distribution or GLT-1: mitochondrial co-localization. We conclude that the association between GLT-1 and mitochondria is modest, is driven by synaptic activity and dependent on polymerized actin filaments. Mitochondria have limited co-localization with the glutamate transporter GLT-1 in primary astrocytes in culture. Few mitochondria are in the fine processes where GLT-1 is abundant. It is necessary to culture astrocytes with neurones to drive a significant level of co-localization, but co-localization is not further altered by depolarization, manipulating sodium ion gradients or Na/K ATPase activity.en_US
dc.language.isoenen_US
dc.relation.isreferencedbyhttp://doi.org/10.1111/jnc.12759en_US
dc.rights© 2014 The Authors. Published Open Access by Wiley. Reproduced in accordance with the publisher's self-archiving policy.en_US
dc.subjectEAAT2; Glial cell; TRAK2; Co-culture; Glutamate transporter; rho kinaseen_US
dc.titleNeuronal influences are necessary to produce mitochondrial co-localization with glutamate transporters in astrocytes.en_US
dc.status.refereedyesen_US
dc.typeArticleen_US
dc.type.versionpublished version paperen_US
refterms.dateFOA2018-07-25T14:17:29Z


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