gabaa receptor subunit a2se  (Addgene inc)

Journal: Cell reports

Article Title: Acute reorganization of postsynaptic GABA A receptors reveals the functional impact of molecular nanoarchitecture at inhibitory synapses.

doi: 10.1016/j.celrep.2023.113331

Figure Lengend Snippet: Figure 2. CRY2olig-GephFingR clustering perturbs the nanodomain organization of postsynaptic scaffolds and receptors (A) Experimental timeline and representative images for live-cell and retrospective dSTORM imaging protocol. Cultured cells expressing CRY2olig-GephFingR- GFP or GephFingR-GFP were imaged live before (dark) and 10 min after light exposure (+light). Cover slips were then fixed and labeled for endogenous gephyrin and GABAARg2, and the same cells were imaged by dSTORM. dSTORM localization data for gephyrin (magenta) and GABAARg2 (blue) are shown to the right, overlayed on the confocal image of CRY2olig-GephFingR. Scale bar represents 2.5 mm. An individual synapse (white box) is magnified to the right. The 2D segmentation image (bottom) displays boundaries defined by minimum localization densities (light shaded areas) and high-density regions (darker shaded re- gions) for gephyrin (magenta) and GABAARg2 (blue). Scale bar represents 250 nm. (B) High-resolution dSTORM example images of inhibitory synapses expressing CRY2olig-GephFingR-GFP or GephFingR-GFP after light-induced clustering. Scale bar represents 200 nm. (C) Left, diagram showing the delineation of GABAAR and gephyrin high-density regions (HDRs) (darker shades) and postsynaptic compartment regions (lighter shades) in dSTORM reconstructions. Middle, mean compartment area of gephyrin and GABAARg2 at synapses in cells expressing CRY2olig-GephFingR-GFP or GephFingR-GFP after light-induced clustering. Error bars represent 95% CI. ***p < 0.001; ****p < 0.0001; Mann-Whitney test. n = 63–70 synapses, from three independent cultures. Right, plotted is the the GABAAR compartment area, measured by dSTORM, for synapses from cells expressing CRY2olig-GephFingR-GFP

Article Snippet: The original GABAA receptor subunit a2SE was a gift from Tija Jacob & Stephen Moss42 (Addgene plasmid # 49169).

Techniques: Imaging, Cell Culture, Expressing, Labeling, MANN-WHITNEY

Journal: Cell reports

Article Title: Acute reorganization of postsynaptic GABA A receptors reveals the functional impact of molecular nanoarchitecture at inhibitory synapses.

doi: 10.1016/j.celrep.2023.113331

Figure Lengend Snippet: Figure 4. CRY2olig-GephFingR clustering impairs evoked and spontaneous synaptic transmission through GABAAR displacement within the postsynaptic membrane (A) Plotted are electrically evoked IPSC amplitudes measured from individual sweeps before and after 2-s light exposures at 0 min and 5 min (denoted by blue bars) recorded from a dissociated hippocampal neuron expressing CRY2olig-GephFingR-GFP. Series resistance measurements are plotted above. (B) Averaged evoked IPSC amplitudes are plotted as a function of time before and after blue light illumination (2-s pulse, denoted by blue arrows) in dissociated hippocampal neurons expressing GephFingR-GFP (black) or CRY2olig-GephFingR-GFP (blue). Error bars are 95% CI. Example IPSC traces before (red) and 10 min following light exposure (blue) are shown above. Scale bar left, 250 pA, 35 ms. Scale bar right, 150 pA, 45 ms. *p < 0.05; two-way ANOVA. n = 7 cells each condition, from 5 independent cultures.

Article Snippet: The original GABAA receptor subunit a2SE was a gift from Tija Jacob & Stephen Moss42 (Addgene plasmid # 49169).

Techniques: Transmission Assay, Membrane, Expressing