Journal: Journal of Neuroinflammation

Article Title: The dopamine analogue CA140 alleviates AD pathology, neuroinflammation, and rescues synaptic/cognitive functions by modulating DRD1 signaling or directly binding to Abeta

doi: 10.1186/s12974-024-03180-x

Figure Lengend Snippet: CA140 improves long-term memory and dendritic spine number in 5xFAD mice. A , B Y-maze and NOR tests of 8-month-old 5xFAD mice injected with vehicle (10% DMSO) or CA140 (30 mg/kg, i.p.) daily for 17 days (n = 10 mice/group). C , D Representative hippocampal AO and BS dendrites of 8-month-old 5xFAD mice injected with vehicle (10% DMSO) or 30 mg/kg CA140 daily for 14 days (AO: Veh, n = 39 neurons from 5 mice; CA140, n = 35 neurons from 5 mice; BS: Veh, n = 35 neurons from 5 mice; CA140, n = 37 neurons from 5 mice). Scale bar = 10 μm. E Measurement of LTP in ex vivo hippocampal slices from 8-month-old 5xFAD mice injected with vehicle (10% DMSO) or CA140 (30 mg/kg, i.p.) daily for 14 days. Representative excitatory postsynaptic current (EPSC) traces before and after long-term potentiation (LTP) induction are shown (1: before LTP; 2: after LTP). F Effects of CA140 on LTP induction in 8-month-old 5xFAD mice. G Summary statistics for LTP induction (last 5 min: Veh, n = 26 cells from 7 mice; CA140, n = 23 cells from 7 mice). H Representative traces of mEPSCs of 6-month-old 5xFAD mice injected with vehicle (10% DMSO) or CA140 (30 mg/kg, i.p.) daily for 14 days. I , J Summary graphs of mEPSC amplitude and frequency in the vehicle and CA140-treated groups (Veh, n = 19 cells from 4 mice; CA140, n = 24 cells from 5 mice). K – N Eight-month-old 5xFAD mice were injected with vehicle (10% DMSO) or CA140 (30 mg/kg, i.p.) daily for 14 days, and immunofluorescence staining was conducted with anti-synaptophysin ( K , L ) or anti-PSD-95 antibodies ( M , N ) (n = 16 brain slices from 4 mice/group). Scale bar = 200 μm. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: PSD-95 , Mouse , 1:200 , Neuromab , 75028 , IF, ICC.

Techniques: Injection, Ex Vivo, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: The dopamine analogue CA140 alleviates AD pathology, neuroinflammation, and rescues synaptic/cognitive functions by modulating DRD1 signaling or directly binding to Abeta

doi: 10.1186/s12974-024-03180-x

Figure Lengend Snippet: CA140 enhances dendritic spine formation by regulating functional synapses in primary hippocampal neurons (PHNs). A , C Dendritic spine density was measured in GFP-transfected PHNs treated with vehicle (1% DMSO) or CA140 (1 or 5 μM) for 24 h on DIV14 and DIV21. B , D Quantification of data from A and C (DIV14: Veh, n = 30; 1 μM CA140, n = 47; 5 μM CA140, n = 48; DIV21: Veh, n = 161; 1 μM CA140, n = 60; 5 μM CA140, n = 50). E–H PHNs were transfected with GFP for 24 h, treated with vehicle (1% DMSO) or CA140 (5 μM) for 24 h, and immunostained with anti-synaptophysin ( E ) or anti-PSD-95 ( G ) antibodies (synaptophysin intensity: Veh, n = 11; CA140, n = 12; synaptophysin puncta number: Veh, n = 55; CA140, n = 42; PSD-95 intensity: Veh, n = 16; CA140, n = 16; PSD-95 puncta number: Veh, n = 15; CA140, n = 14). Scale bar = 20 μm. ** p < 0.01, *** p < 0.001

Article Snippet: PSD-95 , Mouse , 1:200 , Neuromab , 75028 , IF, ICC.

Techniques: Functional Assay, Transfection

Journal: The Journal of Cell Biology

Article Title: Dynein light chain regulates axonal trafficking and synaptic levels of Bassoon

doi: 10.1083/jcb.200807155

Figure Lengend Snippet: Mapping of the DLC-binding region on Bassoon. (A) Overview of Bassoon fragments tested for binding to DLC1 in yeast two-hybrid assays. The extension of positive Bsn clones is displayed in black, and nonbinding clones are indicated in gray. (B) The homologous region of Piccolo does not bind DLC. Amino acid residues encoded by individual subclones are given in parentheses. Zn1/2, zinc fingers; CC1–3, coiled-coil regions; PDZ, PSD-95/Dlg/zonula occludens-1 homology domain; C2A/B, C2 domains; PBH1–10, regions of the Piccolo-Bassoon homology domain (depicted as gray boxes).

Article Snippet: Mouse antibodies used were against cytochrome c (diluted 1:100 for ICC; BD), IC74 (diluted 1:2,000 for WBs and 1:50 for ICC; Millipore), Flag M2 (diluted 1:10,000 for WBs; Sigma-Aldrich), kinesin heavy chain (H2; diluted 1:2,000 for WBs; Millipore), c-myc (9E10; diluted 1:1,000 for WBs; Santa Cruz Biotechnology, Inc.), α-tubulin (diluted 1:1,000; Sigma Aldrich), synapsin (diluted 1:2,000 for ICC; Synaptic Systems GmbH), PSD-95 (diluted 1:1,000–2,000 for ICC; Millipore), and Bassoon (mab7f; diluted 1:2,000 for WBs and 1:1,000 for ICC; Assay Designs).

Techniques: Binding Assay, Clone Assay, Zinc-Fingers

Journal: The Journal of Cell Biology

Article Title: Dynein light chain regulates axonal trafficking and synaptic levels of Bassoon

doi: 10.1083/jcb.200807155

Figure Lengend Snippet: Overexpression of dominant-negative fragment Bsn9 enhances accumulation of Bassoon and Piccolo at synapses. (A) Cells transfected at 3 DIV with GFP-Bsn (top) and GFP-BsnDBM (bottom) were fixed at 15 DIV and counterstained to visualize synapsin (last image in each row and magenta in merged image) and PSD-95 (first image in each row and magenta in merged image). The fluorescence of GFP fusion proteins is shown in the middle image of each row (green in merged images). The second and fourth panels are merged images of panels to their right and left. Higher magnifications of the boxed regions are shown below the respective image. (B–G) Quantification of relative staining intensities for Bassoon (B), Piccolo (C), Munc13 (D), PSD-95 (E), synaptophysin (F), and vGLUT1 (G) in cells transfected at 3 DIV with the bicistronic construct expressing EGFP-synapsin and Bsn9 (red bar) or control fragment Bsn9 II,III (open bar) and analyzed at 9 DIV. Bar graphs show mean values of relative fluorescence intensity for each staining, and error bars indicate SEM. Values are derived from three independent experiments. ***, P < 0.001. Bars: (A) 20 µm; (insets) 10 µm.

Article Snippet: Mouse antibodies used were against cytochrome c (diluted 1:100 for ICC; BD), IC74 (diluted 1:2,000 for WBs and 1:50 for ICC; Millipore), Flag M2 (diluted 1:10,000 for WBs; Sigma-Aldrich), kinesin heavy chain (H2; diluted 1:2,000 for WBs; Millipore), c-myc (9E10; diluted 1:1,000 for WBs; Santa Cruz Biotechnology, Inc.), α-tubulin (diluted 1:1,000; Sigma Aldrich), synapsin (diluted 1:2,000 for ICC; Synaptic Systems GmbH), PSD-95 (diluted 1:1,000–2,000 for ICC; Millipore), and Bassoon (mab7f; diluted 1:2,000 for WBs and 1:1,000 for ICC; Assay Designs).

Techniques: Over Expression, Dominant Negative Mutation, Transfection, Fluorescence, Staining, Construct, Expressing, Derivative Assay