mouse monoclonal antibody against synapsin  (Developmental Studies Hybridoma Bank)

Journal: Frontiers in Cellular Neuroscience

Article Title: A Purinergic P2 Receptor Family-Mediated Increase in Thrombospondin-1 Bolsters Synaptic Density and Epileptic Seizure Activity in the Amygdala-Kindling Rat Model

doi: 10.3389/fncel.2018.00302

Figure Lengend Snippet: Increased synaptic density and transforming growth factor-β1 (TGF-β1) during epileptic progression. The represented changes in IR of related proteins that begin at different stages in different subregions are presented. Measurements of synapsin-I-IR in the ipsilateral hippocampus (Hip), PC and amygdala (AM) in stage 2 (A) and bilateral hippocampus (H), piriform cortex (P) and cortex except PC (C) in stage 4 ( B ; i-ipsilateral; c-contralateral) by western blotting during epileptic progression ( n = 4 per group). Ipsilateral IR of post synaptic density protein-95 (PSD-95) and vesicular glutamate transporter-1 (vGluT-1) in stage 2 and 4 ( C , no changes, –; increase, ↑; n = 4 per group). Flow cytometry-based quantification of TGF-β1 ( D , n = 4 per group). The bands were excised from different gels, which were run under the same electrophoresis condition. Data are shown as mean ± SEM. ** P < 0.01 and *** P < 0.001 compared with controls.

Article Snippet: After blocking with 5% skimmed milk for 1 h, the membranes were incubated with mouse monoclonal antibody against synapsin-I (1:1,000, Abcam, ab8), PSD-95 (1:1,000; Abcam, ab2723), vesicular glutamate transporter-1 (vGluT-1, 1:1,000; Abcam, ab106289), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, Goodhere, AB-P-R-001) at 4°C overnight.

Techniques: Western Blot, Flow Cytometry, Electrophoresis

Journal: Frontiers in Cellular Neuroscience

Article Title: A Purinergic P2 Receptor Family-Mediated Increase in Thrombospondin-1 Bolsters Synaptic Density and Epileptic Seizure Activity in the Amygdala-Kindling Rat Model

doi: 10.3389/fncel.2018.00302

Figure Lengend Snippet: Inhibitory effects of the leu-ser-lys-leu (LSKL) peptide delivered to the lateral ventricle on synapses and kindling progression. (A) IR of PSD-95 (green, sham group, without kindling; bar = 10 μm), (B,C) the ipsilateral IR of synapsin-I/vGluT-1 after different doses of LSKL (H, hippocampus; P, piriform cortex; C, cortex except PC), (D) number of PSD-95-positive puncta in the field (80 μm × 60 μm), (E) Racine stage progression of epilepsy, (F) seizure stages after 20 stimulations, (G) generalized seizure duration after 20 stimulations, (H) afterdischarge duration (ADD), (I) changes in AD threshold (ADT) and (J) respective electroencephalograms (EEGs) recorded from the amygdala in kindled rats (saline group, n = 10; LSKL groups, n = 8–10 per group). The bands were excised from different gels, which were run under the same electrophoresis condition. Data are shown as mean ± SEM. Asterisks show significant differences from the control group (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: After blocking with 5% skimmed milk for 1 h, the membranes were incubated with mouse monoclonal antibody against synapsin-I (1:1,000, Abcam, ab8), PSD-95 (1:1,000; Abcam, ab2723), vesicular glutamate transporter-1 (vGluT-1, 1:1,000; Abcam, ab106289), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, Goodhere, AB-P-R-001) at 4°C overnight.

Techniques: Electrophoresis

Journal: Frontiers in Cellular Neuroscience

Article Title: A Purinergic P2 Receptor Family-Mediated Increase in Thrombospondin-1 Bolsters Synaptic Density and Epileptic Seizure Activity in the Amygdala-Kindling Rat Model

doi: 10.3389/fncel.2018.00302

Figure Lengend Snippet: Effect of pyridoxalphosphate-6-azophenyl-2’,4’-disulfonic (PPADS) on TSP-1, synapses and kindling progression. (A) IR of TSP-1 (red), GFAP (green, bar = 200 μm), and PSD-95 (green, sham group, without kindling; bar = 10 μm) in the dentate gyrus of the hippocampus, (B) mean intensity of TSP-1 immunostaining, (C) number of PSD-95-positive puncta in the field (80 μm × 60 μm), (D) the ipsilateral IR of synapsin-I after different doses of PPADS (H, hippocampus; P, piriform cortex; C, cortex except PC), (E) ADD, (F) seizure stage after 19 stimulations, (G) duration of generalized seizures, (H) Racine stage progression of epilepsy, (I) change in the ADT, and (J) respective EEGs in kindled rats (saline group, n = 10; PPADS groups, n = 8–10 per group). The bands were excised from different gels, which were run under the same electrophoresis condition. DAPI, blue. Data are shown as mean ± SEM. Asterisks show significant differences from the control group (* P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: After blocking with 5% skimmed milk for 1 h, the membranes were incubated with mouse monoclonal antibody against synapsin-I (1:1,000, Abcam, ab8), PSD-95 (1:1,000; Abcam, ab2723), vesicular glutamate transporter-1 (vGluT-1, 1:1,000; Abcam, ab106289), or glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:2,000, Goodhere, AB-P-R-001) at 4°C overnight.

Techniques: Immunostaining, Electrophoresis