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Probing the ionotropic activity of glutamate GluD2 receptor in HEK cells with genetically-engineered photopharmacology

Abstract : Glutamate delta (GluD) receptors belong to the ionotropic glutamate receptor family, yet they don't bind glutamate and are considered orphan. Progress in defining the ion channel function of GluDs in neurons has been hindered by a lack of pharmacological tools. Here, we used a chemo-genetic approach to engineer specific and photo-reversible pharmacology in GluD2 receptor. We incorporated a cysteine mutation in the cavity located above the putative ion channel pore, for site-specific conjugation with a photoswitchable pore blocker. In the constitutively open GluD2 Lurcher mutant, current could be rapidly and reversibly decreased with light. We then transposed the cysteine mutation to the native receptor, to demonstrate with high pharmacological specificity that metabotropic glutamate receptor signaling triggers opening of GluD2. Our results assess the functional relevance of GluD2 ion channel and introduce an optogenetic tool that will provide a novel and powerful means for probing GluD2 ionotropic contribution to neuronal physiology.
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https://hal.archives-ouvertes.fr/hal-03087878
Contributor : Philippe Faure Connect in order to contact the contributor
Submitted on : Friday, October 15, 2021 - 4:44:15 PM
Last modification on : Tuesday, October 19, 2021 - 2:51:54 PM

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Damien Lemoine, Sarah Mondoloni, Jérome Tange, Bertrand Lambolez, Philippe Faure, et al.. Probing the ionotropic activity of glutamate GluD2 receptor in HEK cells with genetically-engineered photopharmacology. eLife, eLife Sciences Publication, 2020, 9, pp.e59026. ⟨10.7554/eLife.59026⟩. ⟨hal-03087878⟩

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