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Arachidonic acid elicits a substrate-gated proton current associated with the glutamate transporter EAAT4

Nature Neuroscience volume 1, pages 105–113 (1998)Cite this article

Abstract

Arachidonic acid modulates both electrical and biochemical properties of membrane proteins involved in cellular signaling. In Xenopus laevis oocytes expressing the excitatory amino acid transporter EAAT4, physiologically relevant concentrations of arachidonic acid increase the amplitude of the substrate-activated current by roughly twofold at -60 mV. This stimulation is not attributable to the modulation of either substrate/ion cotransport or the ligand-gated chloride current, the major conductance associated with this carrier. Ion-substitution experiments reveal that arachidonic acid stimulates a proton-selective conductance. The effect does not require metabolism of arachidonic acid and is not blocked by inhibitors of endogenous oocyte ion-exchangers. This proton conductance expands the complex repertoire of the ligand-gated channel properties associated with EAAT4.

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Figure 1: Representative traces of the effects of L-Asp, arachidonic acid and L-Asp plus arachidonic acid on currents in water-injected and EAAT4-expressing Xenopus oocytes voltage-clamped at -60 mV.
Figure 2: Concentration dependence of the stimulation of the L-Asp-induced current by arachidonic acid and the effects of PUFAs on the L-Asp-induced current in EAAT4-expressing oocytes voltage-clamped at -60 mV.
Figure 3: Effects of arachidonic acid on the concentration response curves for [3H]L-Asp flux and L-Asp-induced currents.
Figure 4: Representative current–voltage curves for L-Asp and arachidonic acid in EAAT4-expressing and uninjected Xenopus oocytes.
Figure 5: Influence of ion substitutions on L-Asp-induced currents and arachidonic acid-stimulated L-Asp currents.
Figure 6: Dependence on pH of the L-Asp-induced and the arachidonic acid-stimulated current in chloride-depleted conditions.
Figure 7: Effects of D2O substitution on translocation and EAAT4 currents.

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Acknowledgements

This work was supported by the Howard Hughes Medical Institute and NIH grant NS33273 to S.G.A. We thank Mary Oltman for technical assistance.

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Authors and Affiliations

  1. Howard Hughes Medical Institute,, Portland, 97201, Oregon, USA

    Wendy A. Fairman & Susan G. Amara

  2. Vollum Institute for Advanced Biomedical Research, Portland, 97201, Oregon, USA

    Wendy A. Fairman, Mark S. Sonders & Susan G. Amara

  3. Department of Pathology, Oregon Health Sciences University, Portland, 97201, Oregon, USA

    Geoffrey H. Murdoch

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  1. Wendy A. Fairman
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Correspondence to Susan G. Amara.

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Fairman, W., Sonders, M., Murdoch, G. et al. Arachidonic acid elicits a substrate-gated proton current associated with the glutamate transporter EAAT4. Nat Neurosci 1, 105–113 (1998). https://doi.org/10.1038/355

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