Journal:

Article Title: Long Term Synaptic Depression That Is Associated with GluR1 Dephosphorylation but Not ?-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid (AMPA) Receptor Internalization *

doi: 10.1074/jbc.M803431200

Figure Lengend Snippet: Neither GluR1 nor GluR2/3 total surface expression is changed 45 min following cLTD treatment, as determined by cross-linking assay. A, representative blots of X-linked (25.0 μg of total protein loaded) and un-X-linked (5.0 μg of total protein loaded) samples for control and cLTD conditions probed with antibodies specific for GluR1 (left group) and GluR2/3 (right group). B, representative blots of un-X-linked and X-linked (5.0μg of total protein loaded for both) samples probed with antibodies specific for PP2Aα (top bands) and synapsin (bottom bands). C and D, graphs of cumulative data comparing X-linked control and X-linked cLTD samples probed with antibodies specific for GluR1 (C) and GluR2/3 (D)(n = 8). E and F, data from C and D normalized to the transferrin receptor. G and H, graphs of cumulative data comparing the percent surface expression (see “Experimental Procedures” for explanation of calculation) between control and cLTD conditions for GluR1 (G) and GluR2/3 (H)(n = 8). I and J, graphs of cumulative data comparing un-X-linked and X-linked samples probed with antibodies specific for PP2Aα (I) and synapsin (J)(n = 8). No significant differences were observed for any of the comparisons as determined by Student's paired t test. Values represent the mean ± S.E.

Article Snippet: Primary antibodies were as follows: GluR1 and GluR2/3 (Chemicon) used at 1:3000 in BSA, phospho-Ser-845 and phospho-Ser-831 (PhosphoSolutions) used at 1:1000 in BSA, PP2Aα (BD Pharmingen) used at 1:5000 in milk, synapsin, NR2B, and NR1 (PhosphoSolutions) used at 1:2000 in BSA, the α subunit of calcium/calmodulin-dependent protein kinase II (αCaMKII) (BD Pharmingen) used at 1:3000 in BSA, phospho-Thr-286 (PhosphoSolutions) used at 1:3000 in milk, PSD-95 (Affinity Bioreagents) used at 1:5000 in BSA, and transferrin receptor (Zymed Laboratories Inc.) used at 1:1000 in milk.

Techniques: Expressing, Control

Journal: International journal of molecular sciences

Article Title: Treadmill Exercise Alleviates Cognition Disorder by Activating the FNDC5: Dual Role of Integrin αV/β5 in Parkinson's Disease.

doi: 10.3390/ijms24097830

Figure Lengend Snippet: Figure 6. Effect of treadmill exercise on synaptic plasticity of hippocampal pyramidal neurons and dopamine levels and BDNF in MPTP-induced chronic PD mice model. (A) Total dopamine levels of the hippocampus (F(4,55) = 33.12, p < 0.001, n = 12). (B) Hippocampal protein levels of PSD95, synapsin, synaptophysin, SNAP47, TH, and BDNF were examined by Western blotting. (C–H) Quantification of PSD95 (F(4,10) = 53.50, p < 0.001), synapsin (F(4,10) = 99.18, p < 0.001), SNAP47 (F(4,10) = 60.12, p < 0.001), synaptophysin (F(4,10) = 17.47, p < 0.001), TH (F(4,10) = 83.36, p < 0.001),and BDNF (F(4,10) = 215.7, p < 0.001) levels. (n = 3). (I) Ultrastructure of synapses determined via electron micrography in hippocampus CA1 region of mice (scale bar = 500 nm). Enlarged images of the left images (scale bar = 125 nm). Red arrows indicate postsynaptic densities (PSDs), green arrows indicate the width of the synaptic cleft, blue arrows indicate the thickness of the PSD, and red dotted lines indicate the length of active synaptic zone. (J–L) Quantification of the length of the active synaptic zone (F(2,6) = 264.3, p < 0.001), synaptic cleft width (F(2,6) = 23.87, p = 0.001), and PSD thickness (F(2,6) = 21.17, p = 0.002), (n = 3). (M,N) Representative section of Golgi-Cox staining dendrites of CA1 pyramidal neurons from the stratum radiatum in mice. Spines number/10 µm (F(2,6) = 11.73, p = 0.009). Scale bars = 10 µm, 25 µm, and 100 µm; n = 3. Data represent the mean ± SEM; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs.

Article Snippet: These primary antibodies included TH (1:1000, ab112, Abcam, Shanghai, China), PSD95 (1:1000, 3450S, CST, Fall River, MA, USA), Synapsin (1:1000, NB300-104, NOVUS (Shanghai) Co. Ltd, Centennial, CO, USA), CAMKII(1:1000, 50049S, CST, Fall River, MA, USA), SNAP47 (1:1000, ab172609, Abcam, Shanghai, China), Synaptophysin (1:1000, ab52636, Abcam, Shanghai, China), BDNF (1:200, ANT-010, Alomone Labs, Jerusalem, Israel), FNDC5 (1:1000, bs-8486R, Bioss, Beijing, China), integrin αV (1:200, sc-9969, Santa Cruz, Dallas, TX, USA), integrin β5 (1:200, sc398214, Santa Cruz, Dallas, TX, USA), and β-actin (1:5000, 60008-1-Ig, Proteintech, Wuhan, China).

Techniques: Western Blot, Staining, Control

Journal: Science Advances

Article Title: Loss of intracellular ATP affects axoplasmic viscosity and pathological protein aggregation in mammalian neurons

doi: 10.1126/sciadv.adq6077

Figure Lengend Snippet: ( A ) Liquid phase–separated condensates formed after 1 hour at 37°C from 10 μM protein in Tyrode’s solution and without crowding agents. Scale bar, 1 μm. ( B ) Estimated size of protein condensates obtained from binary images [ n = 278, 151, 247, 280, and 268 aggregates for SYN1, SNCA, SNCA A53T mutant, Tau (TAU), and TDP-43, respectively]. Box plot with median and minimum/maximum whiskers. Kruskal-Wallis nonparametric test with Dunn’s correction for multiple comparison. ** P = 0.0064, *** P = 0.0002, and **** P < 0.0001. ( C ) Proportion of remaining condensates (left) and size of remaining condensates (right) after adding increasing concentrations of ATP ranging from 2 to 8 mM. Means ± SEM (pooled from four independent experiments). IC 50 , half-maximal inhibition concentration. ( D ) Representative images of SNCA (top) and SNCA A53T mutant (bottom) protein condensates and prefibrils after 1 to 3 hours of incubation at 37°C and after decondensation by 3.2 and 6.4 mM ATP, respectively. Scale bar, 5 μm. ( E ) Proportion of SNCA and SNCA A53T condensates (left; n = 202 and 244 condensates, respectively) and prefibrils (right; n = 4 and 11 prefibrils, respectively) after 3 hours of incubation at 37°C. Data collected and pooled from four independent experiments. Unpaired Student’s t test. ** P = 0.0038.

Article Snippet: Human recombinant full length protein SYN1 (#TP321273) was purchased from OriGene Technologies Inc. Human recombinant full-length proteins SNCA (#pro-393), SNCA A53T mutant (#pro-159), microtubule-associated protein Tau (#pro-295), ubiquitin ligase Parkin (PARK2, #pro-1840), APP N-terminal domain (#pro-1080), TAR DNA binding protein (TDP-43, #pro-2383), and RNA binding protein LIN28 (#pro-743) were purchased from ProSpec.

Techniques: Mutagenesis, Comparison, Inhibition, Concentration Assay, Incubation

Journal: Science Advances

Article Title: Loss of intracellular ATP affects axoplasmic viscosity and pathological protein aggregation in mammalian neurons

doi: 10.1126/sciadv.adq6077

Figure Lengend Snippet: ( A ) Representative images of liquid phase–separated SYN1 condensates formed in presence of 5% PEG and after 60′ without PEG at 37°C from 10 μM protein solutions under control condition (top) or after the addition of 8 mM ATP (bottom). Scale bar, 5 μm. ( B ) Distribution of SYN1 droplet sizes with and without PEG. Means ± SEM (pooled from four independent experiments), multiple unpaired Student’s t test. * P = 0.0061, 0.0326, 0.0482, and 0.0265. ( C ) Average size of SYN1 droplets with and without PEG. Means ± SEM (pooled from four independent experiments), unpaired Student’s t test. * P = 0.0258. ( D ) Normalized number of SYN1 droplets with and without PEG under control conditions or after the addition of ATP. Means ± SEM (pooled from four independent experiments), unpaired Student’s t test. ** P = 0.0049 and **** P < 0.0001. ( E ) Normalized size of SYN1 droplets with and without PEG under control conditions or after the addition of ATP. Means ± SEM (pooled from four independent experiments), unpaired Student’s t test. * P = 0.0182 and **** P < 0.0001.

Article Snippet: Human recombinant full length protein SYN1 (#TP321273) was purchased from OriGene Technologies Inc. Human recombinant full-length proteins SNCA (#pro-393), SNCA A53T mutant (#pro-159), microtubule-associated protein Tau (#pro-295), ubiquitin ligase Parkin (PARK2, #pro-1840), APP N-terminal domain (#pro-1080), TAR DNA binding protein (TDP-43, #pro-2383), and RNA binding protein LIN28 (#pro-743) were purchased from ProSpec.

Techniques: Control