anti glua1  (Santa Cruz Biotechnology)

Journal: Journal of Neurophysiology

Article Title: Two-stage AMPA receptor trafficking in classical conditioning and selective role for glutamate receptor subunit 4 (tGluA4) flop splice variant

doi: 10.1152/jn.01097.2011

Figure Lengend Snippet: Confocal images of abducens motor neurons after normal conditioning for 2 pairing sessions (C2) or conditioning following treatment with the glutamate receptor A4 (GluA4) subunit small-interfering RNA (tGluA4 siRNA). Images are unprocessed except that the contrast was increased for the illustration. After drawing the outline of the cell of interest by the investigator, the software breaks the original image into its individual color channels, revealing punctate staining for synaptophysin (Syn; green), GluA4 (red), or GluA1 (blue) for quantitative analysis. Original scale bar = 10 μm.

Article Snippet: The GluA1 siRNA was obtained from commercial suppliers (sc-35485, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Small Interfering RNA, Software, Staining

Journal: Journal of Neurophysiology

Article Title: Two-stage AMPA receptor trafficking in classical conditioning and selective role for glutamate receptor subunit 4 (tGluA4) flop splice variant

doi: 10.1152/jn.01097.2011

Figure Lengend Snippet: α-Amino-3-hydroxy-5-methylisoxazole-4-propionic acid type receptor (AMPAR) subunit protein expression in brain stem preparations from the different treatment groups. Some preparations were either given no stimulation (Naïve) or underwent pseudoconditioning (Ps2) or conditioning (C2) without further treatment. Other preparations were incubated in siRNA alone, followed by conditioning [tGluA4 siRNA, GluA1 siRNA, negative control (NC) siRNA]. Finally, preparations were injected with rescue plasmids into the abducens nerve for transfection of abducens motor neurons, followed by anti-tGluA4 siRNA treatment and C2 (rescue tGluA4 flip; rescue tGluA4 flop). Actin-loading controls are also shown. *Significant differences from Ps2; #significant differences from C2.

Article Snippet: The GluA1 siRNA was obtained from commercial suppliers (sc-35485, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Expressing, Incubation, Negative Control, Injection, Transfection

Journal: Journal of Neurophysiology

Article Title: Two-stage AMPA receptor trafficking in classical conditioning and selective role for glutamate receptor subunit 4 (tGluA4) flop splice variant

doi: 10.1152/jn.01097.2011

Figure Lengend Snippet: Acquisition of conditioning is inhibited by anti-tGluA4 or anti-GluA1 siRNA treatment and rescued by tGluA4 flop rescue plasmid. Percent of conditioned response (CR) acquisition after 1 and 2 pairing sessions for the different treatment groups is shown. Treated preparations were incubated in the siRNA alone followed by conditioning or pretreated with rescue plasmids followed by tGluA4 siRNA and the conditioning procedure. Physiological traces from some of these groups show representative recordings of CRs (arrows) and URs during paired stimulation. *Significant differences from Ps2; #significant differences from C2. CS, conditioned stimulus; US, unconditioned stimulus.

Article Snippet: The GluA1 siRNA was obtained from commercial suppliers (sc-35485, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Plasmid Preparation, Incubation

Journal: Journal of Neurophysiology

Article Title: Two-stage AMPA receptor trafficking in classical conditioning and selective role for glutamate receptor subunit 4 (tGluA4) flop splice variant

doi: 10.1152/jn.01097.2011

Figure Lengend Snippet: Synaptic localization of tGluA4- or tGluA1-containing AMPARs after conditioning or treatment with siRNA and rescue plasmids. A: representative confocal images of abducens motor neurons from the different treatment groups showing punctate staining for synaptophysin (green), GluA1 AMPAR subunits (blue), and GluA4 (red). Double colocalization of GluA1 with synaptophysin (GluA1 + Syn) is cyan, GluA4 with synaptophysin (GluA4 + Syn) is yellow, and triple colocalization of GluA1 and GluA4 with synaptophysin (GluA1 + GluA4 + Syn) is white. Original scale bar = 2 μm. B: quantitative analysis of synaptophysin punctate staining and the colocalization of AMPAR subunits with synaptophysin for the different treatment groups. *Significant differences from Ps2; #significant differences from C2.

Article Snippet: The GluA1 siRNA was obtained from commercial suppliers (sc-35485, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Staining

Journal: Journal of Neurophysiology

Article Title: Two-stage AMPA receptor trafficking in classical conditioning and selective role for glutamate receptor subunit 4 (tGluA4) flop splice variant

doi: 10.1152/jn.01097.2011

Figure Lengend Snippet: Synaptic localization of tGluA2/3-containing AMPARs after conditioning or treatment with the siRNAs. Levels of synaptophysin punctate staining increased significantly in all of the groups that received paired stimulation compared with Ps2 (P < 0.0001). Conditioning in normal medium or after treatment with either the tGluA4 or GluA1 siRNA did not alter the colocalization of these AMPARs with synaptophysin. Representative images of abducens motor neurons are also shown (synaptophysin, green; tGluA2/3, red; colocalization, yellow). *Significant differences from Ps2; scale bar = 2 μm.

Article Snippet: The GluA1 siRNA was obtained from commercial suppliers (sc-35485, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Staining