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Optical control of metabotropic glutamate receptors

Nature Neuroscience volume 16, pages 507516 (2013) | Download Citation

Abstract

G protein–coupled receptors (GPCRs), the largest family of membrane signaling proteins, respond to neurotransmitters, hormones and small environmental molecules. The neuronal function of many GPCRs has been difficult to resolve because of an inability to gate them with subtype specificity, spatial precision, speed and reversibility. To address this, we developed an approach for opto-chemical engineering of native GPCRs. We applied this to the metabotropic glutamate receptors (mGluRs) to generate light-agonized and light-antagonized mGluRs (LimGluRs). The light-agonized LimGluR2, on which we focused, was fast, bistable and supported multiple rounds of on/off switching. Light gated two of the primary neuronal functions of mGluR2: suppression of excitability and inhibition of neurotransmitter release. We found that the light-antagonized tool LimGluR2-block was able to manipulate negative feedback of synaptically released glutamate on transmitter release. We generalized the optical control to two additional family members: mGluR3 and mGluR6. This system worked in rodent brain slices and in zebrafish in vivo, where we found that mGluR2 modulated the threshold for escape behavior. These light-gated mGluRs pave the way for determining the roles of mGluRs in synaptic plasticity, memory and disease.

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Acknowledgements

We thank A.P. Mariani (National Eye Institute) for 11-cis retinal, J.P. Pin (University of Montpellier) for the mGluR plasmids and E. Reuveny (Weizmann Institute) for the GIRK1 plasmid, K. Durkin, K. Dubay, T. Berger, G. Sandoz and S. Berlin for helpful discussions, A. Guyon, Z. Fu and S. Szobota for help with slice cultures, Z. Fu for molecular biology assistance, K. McDaniel, J. Maxfield, J. Saint-Hillaire and D. Weinman for fish care, E. Carroll for discussion and help with zebrafish set up, P. Gut (University of California, San Francisco) for zebrafish plasmids, H. Baier (University of California, San Francisco) for fish lines, and the College of Chemistry (University of California, Berkeley) for computing resources for the Monte Carlo simulations. Support for the work was provided by the Nanomedicine Development Center for the Optical Control of Biological Function, US National Institutes of Health grant PN2EY018241 (D.T. and E.Y.I.), the Human Frontier Science Program (RGP0013/2010 to E.Y.I.), the Deutsche Forschungsgemeinschaft (FOR 1279, D.T.), the Fond der Chemischen Industrie (Kekulé fellowship to P.S.), National Science Foundation grants CHE-0233882 and CHE-0840505 (to the College of Chemistry at the University of California, Berkeley), a postdoctoral fellowship of the European Molecular Biology Organization (H.J.), a Helen Hay Whitney postdoctoral fellowship (D.S.), the McKnight Endowment Fund for Neuroscience and US National Institutes of Health grant R01HL109525 (A.F.S.), and a predoctoral fellowship from the Fulbright Foundation (B.G.).

Author information

Author notes

    • Harald Janovjak

    Present address: Institute of Science and Technology Austria, Klosterneuburg, Austria.

Affiliations

  1. Biophysics Graduate Group, University of California, Berkeley, California, USA.

    • Joshua Levitz
    •  & Ehud Y Isacoff
  2. Department of Molecular and Cell Biology, University of California, Berkeley, California, USA.

    • Carlos Pantoja
    • , Harald Janovjak
    • , Andreas Reiner
    • , Adam Hoagland
    •  & Ehud Y Isacoff
  3. Helen Wills Neuroscience Graduate Program, University of California, Berkeley, California, USA.

    • Benjamin Gaub
    •  & Ehud Y Isacoff
  4. Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA.

    • David Schoppik
    •  & Alexander F Schier
  5. Department of Chemistry, University of California, Berkeley, California, USA.

    • Brian Kane
    •  & Dirk Trauner
  6. Department of Chemistry and Center of Integrated Protein Science, University of Munich, Munich, Germany.

    • Philipp Stawski
    •  & Dirk Trauner
  7. Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.

    • Ehud Y Isacoff

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Contributions

J.L. designed experiments and conducted homology modeling, Monte Carlo simulations, HEK293 cell patch experiments, cAMP measurements, cultured neuron and hippocampal slice patch experiments, analyzed data, and wrote the manuscript. C.P. designed zebrafish experiments, conducted zebrafish behavioral experiments, analyzed data, and wrote the manuscript. B.G. and A.R. conducted HEK293 cell patch experiments. H.J. designed and conducted Monte Carlo simulations. P.S. and B.K. synthesized photoswitches. A.H., D.S. and A.F.S. developed and setup zebrafish behavioral apparatus. D.T. developed photoswitching methodology and provided photoswitches. E.Y.I. supervised the project, designed experiments, analyzed data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Dirk Trauner or Ehud Y Isacoff.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–10, Supplementary Table 1 and Chemical Synthesis

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DOI

https://doi.org/10.1038/nn.3346

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