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The Journal of Neuroscience, December 1, 1999, 19(23):10193-10200
Glutamate Induces Rapid Upregulation of Astrocyte Glutamate Transport and Cell-Surface Expression of GLAST
Shumin Duan, Christopher M. Anderson, Becky A. Stein, and Raymond A. Swanson Department of Neurology, University of California, San Francisco, and Veterans Affairs Medical Center, San Francisco, California 94121
Glutamate transporters clear glutamate from the extracellular space by high-affinity binding and uptake. Factors that regulate glutamate transporter expression and activity can thereby influence excitatory neurotransmission. Transporter function in GABAergic and other systems has been shown to be regulated by transporter substrates. Here, glutamate regulation of glutamate transport was studied using primary murine astrocyte cultures that express the GLAST (EAAT1) and GLT-1 (EAAT2) transporter subtypes. Glutamate was found to stimulate glutamate transport capacity (Vmax) in a dose- and time-dependent manner. The maximal increase was 100%, with an ED50 of 40 µM glutamate and with onset beginning ~15 min after onset of glutamate exposure. The uptake stimulation was reproduced by D-aspartate, which is also a transporter substrate, but not by nontransported glutamate receptor agonists. Moreover, glutamate incubation did not stimulate transport when performed in a sodium-free medium, suggesting that the stimulatory effect of glutamate is triggered by increased transporter activity rather than receptor activation. Treatment with the actin-disrupting agents cytochalasin B or cytochalasin D prevented the glutamate-induced increase in glutamate uptake. Biotinylation labeling of membrane surface proteins showed that glutamate incubation produced an increase in GLAST expression at the astrocyte cell surface. These results suggest that cell-surface expression of GLAST can be rapidly regulated by glutamate through a process triggered by GLAST activity and involving the actin cytoskeleton. This feedback loop provides a mechanism by which changes in extracellular glutamate concentrations could rapidly modulate astrocyte glutamate transport capacity.
Key words: EAAT1; EAAT2; GLT-1; GLAST; actin; cytochalasin; glutamate uptake; trafficking
Copyright © 1999 Society for Neuroscience 0270-6474/99/192310193-08$05.00/0
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