title: Role of Na,K-ATPase α1 and α2 isoforms in the support of astrocyte glutamate uptake.
abstract: Glutamate released during neuronal activity is cleared from the synaptic space via the astrocytic glutamate/Na(+) co-transporters. This transport is driven by the transmembrane Na(+) gradient mediated by Na,K-ATPase. Astrocytes express two isoforms of the catalytic Na,K-ATPase α subunits; the ubiquitously expressed α1 subunit and the α2 subunit that has a more specific expression profile. In the brain α2 is predominantly expressed in astrocytes. The isoforms differ with regard to Na+ affinity, which is lower for α2. The relative roles of the α1 and α2 isoforms in astrocytes are not well understood. Here we present evidence that the presence of the α2 isoform may contribute to a more efficient restoration of glutamate triggered increases in intracellular sodium concentration [Na(+)]i. Studies were performed on primary astrocytes derived from E17 rat striatum expressing Na,K-ATPase α1 and α2 and the glutamate/Na(+) co-transporter GLAST. Selective inhibition of α2 resulted in a modest increase of [Na(+)]i accompanied by a disproportionately large decrease in uptake of aspartate, an indicator of glutamate uptake. To compare the capacity of α1 and α2 to handle increases in [Na(+)]i triggered by glutamate, primary astrocytes overexpressing either α1 or α2 were used. Exposure to glutamate 200 µM caused a significantly larger increase in [Na(+)]i in α1 than in α2 overexpressing cells, and as a consequence restoration of [Na(+)]i, after glutamate exposure was discontinued, took longer time in α1 than in α2 overexpressing cells. Both α1 and α2 interacted with astrocyte glutamate/Na(+) co-transporters via the 1st intracellular loop.
authors.affiliation: Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Nina B
authors.initials: NB
authors.lastname: Illarionova
authors.forename: Nina B
authors.initials: NB
authors.lastname: Illarionava
authors.affiliation: Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden; Science for Life Laboratory, Department of Cell Physics, Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Hjalmar
authors.initials: H
authors.lastname: Brismar
authors.affiliation: Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anita
authors.initials: A
authors.lastname: Aperia
authors.affiliation: Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Eli
authors.initials: E
authors.lastname: Gunnarson
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism*; D001692:Biological Transport; D027341:Excitatory Amino Acid Transporter 1 / Q000378:metabolism; D027342:Excitatory Amino Acid Transporter 2 / Q000378:metabolism; D015870:Gene Expression; D018698:Glutamic Acid / Q000378:metabolism*; D042541:Intracellular Space / Q000378:metabolism; D007527:Isoenzymes; D011485:Protein Binding; D051381:Rats; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D027341:Excitatory Amino Acid Transporter 1; D027342:Excitatory Amino Acid Transporter 2; D007527:Isoenzymes; C493395:Slc1a2 protein, rat; C493392:Slc1a3 protein, rat; D001224:Aspartic Acid; D018698:Glutamic Acid; D012964:Sodium; C499075:Atp1a1 protein, rat; C498532:Atp1a2 protein, rat; D000254:Sodium-Potassium-Exchanging ATPase