Neuronal influences are necessary to produce mitochondrial co-localization with glutamate transporters in astrocytes.
dc.contributor.author | Ugbode, Christopher I. | * |
dc.contributor.author | Hirst, W.D. | * |
dc.contributor.author | Rattray, Marcus | * |
dc.date.accessioned | 2016-02-23T16:40:36Z | |
dc.date.available | 2016-02-23T16:40:36Z | |
dc.date.issued | 2014-09 | |
dc.identifier.citation | Ugbode 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.uri | http://hdl.handle.net/10454/7811 | |
dc.description | yes | en_US |
dc.description.abstract | Abstract 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.iso | en | en_US |
dc.rights | © 2014 The Authors. Published Open Access by Wiley. Reproduced in accordance with the publisher's self-archiving policy. | en_US |
dc.subject | EAAT2; Glial cell; TRAK2; Co-culture; Glutamate transporter; rho kinase | en_US |
dc.title | Neuronal influences are necessary to produce mitochondrial co-localization with glutamate transporters in astrocytes. | en_US |
dc.status.refereed | yes | en_US |
dc.type | Article | en_US |
dc.type.version | published version paper | en_US |
dc.identifier.doi | https://doi.org/10.1111/jnc.12759 | |
refterms.dateFOA | 2018-07-25T14:17:29Z |