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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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.).
The authors declare no competing financial interests.
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Levitz, J., Pantoja, C., Gaub, B. et al. Optical control of metabotropic glutamate receptors. Nat Neurosci 16, 507–516 (2013). https://doi.org/10.1038/nn.3346
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