Journal: Neuron

Article Title: Architecture of the Mouse Brain Synaptome

doi: 10.1016/j.neuron.2018.07.007

Figure Lengend Snippet: Whole-Brain-Scale Mapping of PSD95 and SAP102 (A) PSD95 (green) and SAP102 (magenta) expression in stitched down-sampled images of five coronal sections (S1–S5). Bregma (β) level is indicated. (B) PSD95 and SAP102 synaptome parameters in ARA anatomical subregions. Parameter units are as follows: density, number of puncta per 100 μm 2 ; size, μm 2 ; intensity, mean gray value per punctum (AU × 10 4 ); colocalization, %. (C) Synaptome maps of delineated regions in five sections. Median punctum density (i), intensity (ii), size (iii), and colocalization (iv) for PSD95 (top) and SAP102 (bottom) are shown. Parameter units: density, number of puncta per 100 μm 2 ; intensity, mean gray value per punctum (AU); size, μm 2 ; colocalization, %. (D) PSD95 punctum intensity in delineated subregions of the hippocampus and cortex. CA1, cornu ammonis; CX, isocortex; DG, dentate gyrus. (E) SAP102 punctum intensity in delineated subregions of hippocampus and cortex. (F) Similarity matrix between pairs of subregions (rows and columns). White lines outline three major blocks (cerebrum, brainstem, and cerebellum). Pink box highlights hippocampal subregions. (G) Dendrogram showing hierarchical organization of subregions based on their similarity. Branch tips represent delineated subregions, colored as in (B).

Article Snippet: Three-way colocalization was then measured by Imaris Triple Colocalization plugin, developed by Michael Adams, using a distance threshold of 600 nm.

Techniques: Expressing

Journal: Neuron

Article Title: Architecture of the Mouse Brain Synaptome

doi: 10.1016/j.neuron.2018.07.007

Figure Lengend Snippet:

Article Snippet: Three-way colocalization was then measured by Imaris Triple Colocalization plugin, developed by Michael Adams, using a distance threshold of 600 nm.

Techniques: Western Blot, Virus, Recombinant, Plasmid Preparation, Sequencing, Software, Comparison

Journal: iScience

Article Title: The non-adrenergic imidazoline-1 receptor protein nischarin is a key regulator of astrocyte glutamate uptake

doi: 10.1016/j.isci.2022.104127

Figure Lengend Snippet: Nischarin mediates glutamate-dependent GLT-1 internalization in astrocytes (A) Surface biotinylation assay showing surface GLT-1 level in astrocytes transfected with GFP and GLT-1a-V5 or GFP-Nisch and GLT-1a-V5 following +/− 100μM glutamate treatment. One-way ANOVA, post hoc Dunnett’s multiple comparison test (n = 4 individual experiments). (B) Proximity ligation assay in DIV14 hippocampal culture. Increased red puncta per nuclei (DAPI stained (blue)) is indicative of increased direct interaction between Nischarin and GLT-1 in hippocampal culture. Glutamate treatment (100μM, 1h) significantly increased GLT-1-Nischarin interaction compared to control. One-way ANOVA, post hoc Tukey’s test (n = 3 individual preparations). (C) Schematic representation of GLT-1BBS construct bound to BTX conjugated Alexa 555 (BTX555). Astrocytes expressing GFP-Nisch and GLT-1aBBS were labeled using BTX555 and dual color live-structured illumination microscopy-monitored trafficking of GLT-1 following glutamate treatment. Merged kymographs of GFP-Nisch vesicle (green) and GLT-1 bound BTX-555 (red) reveal co-localized diagonal trajectory, representing moving vesicles. (D) Quantification of GFP-Nisch and GLT-1aBBS expressing astrocytes treated with 100μM glutamate for 0, 5, 30, and 60min showed increased colocalization between Nisch and GLT-1 compared to untreated controls. p values by unpaired t test, Mann Whitney test (n = 6-14).

Article Snippet: For measuring colocalization, the colocalization plugin of the Metamorph software was used.

Techniques: Surface Biotinylation Assay, Transfection, Comparison, Proximity Ligation Assay, Staining, Control, Construct, Expressing, Labeling, Microscopy, MANN-WHITNEY