Journal:

Article Title: Alterations in AMPA receptor subunits and TARPs in the rat nucleus accumbens related to the formation of Ca 2+ -permeable AMPA receptors during the incubation of cocaine craving

doi: 10.1016/j.neuropharm.2011.01.021

Figure Lengend Snippet: Expression of AMPAR subunits and the TARP γ2 in the NAc postsynaptic density (PSD) fraction after 1 or 45 days of withdrawal from self-administration. Data are presented as mean (± SEM) expressed as percent of saline controls at each withdrawal time. A) GluA1 protein levels in the Coc-SA group were slightly decreased on WD45 compared to the Sal-SA WD45 group. B) Data suggesting that γ2 and GluA1 are lost in parallel during preparation of PSD fractions from the Coc-SA WD45 group. Total γ2 protein decreased slightly in the Coc-SA WD45 group (inset), similar to the decrease in GluA1 shown in panel A, and levels of GluA1 protein and γ2 protein were significantly correlated for individual rats in this group. The dashed lines show confidence interval and the solid line shows best fit. C, D) GluA2 and GluA3 protein levels were unchanged in the PSD fraction. In all panels, blot images from two rats in each group are shown by the two bands directly underneath each bar. For example, in panel A, the top four bands correspond (from left to right) to Sal-SA WD1, Coc-SA WD1, Sal-SA WD45 and Coc-SA WD45 rats. The lower four bands show another set of representative rats from each of these experimental groups.

Article Snippet: 2.8 SDS-PAGE and immunoblotting All samples were heated at 70°C for 10 min in Laemmli sample treatment buffer with 100mM DTT and then processed for SDS-PAGE and immunoblotting as described previously ( Ferrario et al., 2010 ) using the following primary antibodies: GluA1 (1:1000, PA1-37776, Thermo Scientific or 1:1000, AB1504 or MAB2263, Millipore); pS845 GluA1 (1:500, p1160-845, PhosphoSolutions, Aurora, CO; or 1:500, 2491-1, Epitomics, Burlingame, CA); GluA2 (1:200, 75-002, UC Davis/NIH NeuroMab Facility, Davis, CA or 1:1000, AB1768, Millipore); GluA3 (1:500, 3437, Cell Signaling, Danvers, MA); γ2 (1:1000, 1505-STAR, PhosphoSolutions); γ4 (1:1000, AB5795, Millipore); PSD-95 (1:20,000, 75-028, UC Davis/NIH NeuroMab Facility); CaMKIIα (1:20,000, MAB8699, Millipore); CaMKIIβ (1:500, ab34703; AbCam, Cambridge, MA), pCaMKIIα/β (1:5000, p1005-286, PhosphoSolutions); total ERK1/2 (1:10,000, AB3053, Millipore); pERK1/2 (1:10,000, AB3826, Millipore).

Techniques: Expressing, Saline

Journal:

Article Title: Alterations in AMPA receptor subunits and TARPs in the rat nucleus accumbens related to the formation of Ca 2+ -permeable AMPA receptors during the incubation of cocaine craving

doi: 10.1016/j.neuropharm.2011.01.021

Figure Lengend Snippet: Expression of AMPAR subunits and the TARPs γ2 and γ4 in the NAc lysed synaptosomal membrane (LP1) fraction after 45 days of withdrawal from cocaine-administration (Coc-SA WD45) or the last handling session (Han). Data are presented as mean (± SEM) expressed as percent of handled controls. Importantly, Han and Sal-SA groups did not differ on several measures (see Results), including the RI (Fig. 1). A) GluA1 protein levels were significantly increased in the Coc-SA group compared to the Han group. B) Levels of γ2 and γ4 were slightly elevated. C) GluA2 protein levels were unchanged in the Coc-SA group. D) Levels of GluA3 protein were significantly decreased in the Coc-SA group compared to controls. Taken together, these data are consistent with an increase in CP-AMPARs in the Coc-SA WD45 group. In all panels, blot images from two rats in each group are shown by the two bands directly underneath each bar. For example, in panel A, the top two bands correspond (from left to right) to Han and Coc-SA WD45 rats. The lower two bands show another set of representative rats from each of these experimental groups. *p<.05, #p=.05

Article Snippet: 2.8 SDS-PAGE and immunoblotting All samples were heated at 70°C for 10 min in Laemmli sample treatment buffer with 100mM DTT and then processed for SDS-PAGE and immunoblotting as described previously ( Ferrario et al., 2010 ) using the following primary antibodies: GluA1 (1:1000, PA1-37776, Thermo Scientific or 1:1000, AB1504 or MAB2263, Millipore); pS845 GluA1 (1:500, p1160-845, PhosphoSolutions, Aurora, CO; or 1:500, 2491-1, Epitomics, Burlingame, CA); GluA2 (1:200, 75-002, UC Davis/NIH NeuroMab Facility, Davis, CA or 1:1000, AB1768, Millipore); GluA3 (1:500, 3437, Cell Signaling, Danvers, MA); γ2 (1:1000, 1505-STAR, PhosphoSolutions); γ4 (1:1000, AB5795, Millipore); PSD-95 (1:20,000, 75-028, UC Davis/NIH NeuroMab Facility); CaMKIIα (1:20,000, MAB8699, Millipore); CaMKIIβ (1:500, ab34703; AbCam, Cambridge, MA), pCaMKIIα/β (1:5000, p1005-286, PhosphoSolutions); total ERK1/2 (1:10,000, AB3053, Millipore); pERK1/2 (1:10,000, AB3826, Millipore).

Techniques: Expressing, Membrane

Journal: Structure (London, England : 1993)

Article Title: Druggability simulations and X-ray crystallography reveal a ligand-binding site in the GluA3 AMPA receptor N-terminal domain

doi: 10.1016/j.str.2018.10.017

Figure Lengend Snippet: (A-C) Conserved probe binding patterns observed in multiple druggability simulations of GluA1 (A), GluA2 (B) and GluA3 (C) NTD dimers. In contrast to GluA1 and GluA2, GluA3 features a high affinity ligand-binding site at the LL interface (see contact distributions in Figure S4). (D-H) Our crystal structures of the GluA3 NTD reveal unique dimeric states: one with phosphate bound between the pairs of R163 and R184 residues (D), which is comparable to the previously resolved open conformer (PDB ID: 3O21 dimer CD; see Figure S5); a super-open form (E); and a displaced dimer found in the same crystal as the phosphate bound form (G). Bottom views in F and H compare the PO43−-bound structure to the super-open and displaced structures, respectively. Phosphate binding is mimicked by dianionic methylphosphate binding in additional druggability simulations (see Figure S7).

Article Snippet: Our data revealed extensive probe binding specifically at the GluA3 NTD LL dimer interface, a known allosteric site utilized in other receptors, such as mGluRs and natriuretic peptide receptors ( Misono et al., 2011 ; Tsuchiya et al., 2002 ), and by NMDAR NTDs ( Mony et al., 2011 ; Tajima et al., 2016 ).

Techniques: Binding Assay, Ligand Binding Assay

Journal: Structure (London, England : 1993)

Article Title: Druggability simulations and X-ray crystallography reveal a ligand-binding site in the GluA3 AMPA receptor N-terminal domain

doi: 10.1016/j.str.2018.10.017

Figure Lengend Snippet: Druggability of GluA3 NTD LL interface pockets.

Article Snippet: Our data revealed extensive probe binding specifically at the GluA3 NTD LL dimer interface, a known allosteric site utilized in other receptors, such as mGluRs and natriuretic peptide receptors ( Misono et al., 2011 ; Tsuchiya et al., 2002 ), and by NMDAR NTDs ( Mony et al., 2011 ; Tajima et al., 2016 ).

Techniques: Binding Assay

Journal: Structure (London, England : 1993)

Article Title: Druggability simulations and X-ray crystallography reveal a ligand-binding site in the GluA3 AMPA receptor N-terminal domain

doi: 10.1016/j.str.2018.10.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Our data revealed extensive probe binding specifically at the GluA3 NTD LL dimer interface, a known allosteric site utilized in other receptors, such as mGluRs and natriuretic peptide receptors ( Misono et al., 2011 ; Tsuchiya et al., 2002 ), and by NMDAR NTDs ( Mony et al., 2011 ; Tajima et al., 2016 ).

Techniques: Recombinant, Stable Transfection, Expressing, Software