(A) GluD2 is genetically-modified to incorporate a cysteine residue (yellow) at the entrance to the pore, which serves as a handle for the covalent attachment of a synthetic, photoswitchable …
(A) UV-visible spectra of MAGu under 520 nm light (green, mostly trans) or under 390 nm light (pink, mostly cis). (B) Thermal relaxation of cis MAGu in aqueous solution in darkness. Absorbance at …
(A) Left, representative current-voltage relationship for GluD2-A654T Lc, with (blue) and without (black) pentamidine (100 μM). The subtracted current (gray) shows clear inward rectification and …
Related to Figure 2A, B and C.
(A) Currents recorded at −60 mV for A654T and each of the fifteen cysteine mutants (n = 3–8 cells). (B) Pentamidine block for A654T and each of the fifteen cysteine mutants (n = 3–8 cells). Data are …
(A) Representative current traces (Vm = −60 mV) recorded for A654T-I677C in darkness, then under 380 light (violet), upon application of pentamidine or NMDG (blue, 100 μM), and then under 535 nm …
Related to Figure 3A, C and D.
(A) Average current-versus-voltage relationship under 380 and 535 nm light (n = 21). S.E.M is shown is shade, and subtracted current in gray. (B) Left, representative dose-dependent blockade of the …
Related to Figure 4A and B.
(A) Representative DHPG-induced current (measured as indicated by the red arrow) for a MAGu-treated (20 µM, 20 min) cell co-expressing mGlu1b and GluD2-I677C, under 380, 535 and 380 nm light. Note …
Related to Figure 5C and D.
(A) Top, DHPG (200 μM)-induced current in vehicle-treated cerebellar slices were of similar amplitude under 380 and 535 nm light (−800 ± 57 pA vs. −799 ± 57 pA, p>0.999, n = 10). Middle, DHPG (200 …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Cell line (H. sapiens) | HEK tsa201 | Sigma-Aldrich #96121229 | RRID:CVCL_2737 | |
Chemical compound, drug | MAGu | 10.1016/j.neuron.2015.10.026 | Originally named PAG-1c. Custom-synthetized by Enamine, Ukraine | |
Chemical compound, drug | DMSO | Sigma-Aldrich | D2650 | |
Chemical compound, drug | Pentamidine | Sigma-Aldrich | 1504900 | |
Chemical compound, drug | N-methyl-d-glucamine (NMDG) | Sigma-Aldrich | M2004 | |
Chemical compound, drug | (R,S)−3,5-DHPG | Hello-bio | HB0026 | |
Chemical compound, drug | CNQX | Hello-bio | HB0205 | |
Chemical compound, drug | D-APV | Hello-bio | HB0225 | |
Chemical compound, drug | SR 95531 (Gabazine) | Hello-bio | HB0901 | |
Chemical compound, drug | CGP 55845 | Hello-bio | HB0960 | |
Gene (Mus musculus) | Grid2 (glutamate receptor, ionotropic, delta 2) | Genbank | GeneID: 14804 | |
Gene (Rattus norvegicus) | Grm1 (glutamate receptor, metabotropic 1) | Genbank | Gene ID: 24414 | |
Transfected construct (Mus musculus) | pcDNA3-GluD2 | https://doi.org/10.1002/embr.201337371 | NM_008167.3 | |
Transfected construct (Mus musculus) | pRK5-mGlu1b | Laurent Prezeau (IGF, Montpellier, France). | NM_001114330.1 | |
Software, algorithm | R Project for Statistical Computing | http://www.r-project.org/ | RRID:SCR_001905 | |
Software, algorithm | Modeller 9.19 | https://salilab.org/modeller/9.19/release.html | RRID:SCR_008395 | |
Software, algorithm | Smina | https://sourceforge.net/projects/smina/ |