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AMPA receptor-mediated regulation of a Gi-protein in cortical neurons

Nature volume 389, pages 502–504 (1997)Cite this article

A Corrigendum to this article was published on 20 December 2017

This article has been updated

Abstract

Excitatory synaptic transmission in the central nervous system is mediated primarily by the release of glutamate from presynaptic terminals onto postsynaptic channels gated by N -methyl-D-aspartate (NMDA) and α-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors1,2. The myriad intracellular responses arising from the activation of the NMDA and AMPA receptors have previously been attributed to the flow of Ca2+ and/or Na+ through these ion channels1,2,3,4,5,6. Here we report that the binding of the agonist AMPA to its receptor can generate intracellular signals that are independent of Ca2+ and Na+ in rat cortical neurons. In the absence of intracellular Ca2+ and Na+, AMPA, but not NMDA, brought about changes in a guanine-nucleotide-binding protein (Gαi1) that inhibited pertussis toxin-mediated ADP-ribosylation of the protein in an in vitro assay. This effect was observed in intact neurons treated with AMPA as well as in isolated membranes exposed to AMPA, and was also found in MIN6 cells, which express functional AMPA receptors but have no metabotropic glutamate receptors. AMPA also inhibited forskolin-stimulated activity of adenylate cyclase in neurons, demonstrating that Gi proteins were activated. Moreover, both Gβγ blockage and co-precipitation experiments demonstrated that the modulation of the Gi protein arose from the association of Gαi1 with the glutamate receptor-1 (GluR1) subunit. These results suggest that, as well as acting as an ion channel, the AMPA receptor can exhibit metabotropic activity.

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Figure 1: AMPA, added to intact neurons, inhibited the PTX-mediated ADP-ribosylation of Gαi1.
Figure 2: AMPA, added to membranes isolated from unstimulated neurons, reduced the PTX-mediated ADP-ribosylation of Gαi1 and forskolin-stimulated adenylate cyclase activity.
Figure 3: Reversion of AMPA-inhibited ADP-ribosylation of Gαi1 by exogenous Gβγ subunit in neuronal membranes, and the detection of Gαi1 in immunocomplexes precipitated by an anti-GluR1 antibody in neurons exposed to AMPA.
Figure 4: AMPA added to membranes isolated from unstimulated MIN6 cells, inhibited PTX-mediated ADP-ribosylation of Gαi1a, b.

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Change history

  • 20 December 2017

    Please see accompanying Corrigendum (http://doi.org/10.1038/nature25163). Fig. 4 of this Letter presented three events of data duplication. In Fig. 4a the panels in row 1, columns 1 and 7 were the same, in Fig. 4a the panels in row 3, columns 5 and 7 were the same, and in Fig. 4c and e, the panels in columns 1–3 were the same. Given the time elapsed since publication, the original raw data could not be located. The main conclusion illustrated by Fig. 4, however, of dose-dependent (Fig. 4d) and Gβγ-sensitive (Fig. 4f) activation of Gαi1 by AMPA in MIN6 cells, which do not express GluR6 (Fig. 4b), remains unaffected. Authors J.P.D. and P.M., now both retired, could not be reached. The original Letter has not been corrected online.

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Acknowledgements

We thank R. Tremblay, D. Song and S. MacLean for technical support and supplying neuronal and MIN6 cell cultures; R. Ball and A. Shapiro for the EIA; J. Conn for antibodies against mGluR4 and mGluR7; S. Nakanishi and R. Shigemoto for the antibody against mGluR6; S. Seino and J. Miyazaki for MIN6 cells; B. Chakravarthy for comments, and P. R. Walker for suggestions during the preparation of the manuscript. This work was partly supported by a Heart and Stroke Foundation of Ontario grant awarded under the OSCAR program (J.P.D. and Y.Z.W.).

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  1. Cellular Neurobiology Group, Institute for Biological Sciences, National Research Council of Canada, Ottawa, K1A 0R6, Ontario, Canada

    Yizheng Wang, Daniel L. Small, Danica B. Stanimirovic, Paul Morley & Jon P. Durkin

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Correspondence to Yizheng Wang.

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Wang, Y., Small, D., Stanimirovic, D. et al. AMPA receptor-mediated regulation of a Gi-protein in cortical neurons. Nature 389, 502–504 (1997). https://doi.org/10.1038/39062

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