mouse anti psd 95 upstate  (Alomone Labs)

Journal:

Article Title: Persistent inflammation induces GluR2 internalization via NMDA receptor-triggered PKC activation in dorsal horn neurons

doi: 10.1523/JNEUROSCI.4514-08.2009

Figure Lengend Snippet: NMDARs and PKC couple to AMPAR complex in dorsal horn. (A) PICK1 co-immunoprecipitates with GluR2 and PKCα (but not PKCβ or PKCγ). (B) GluR2 co-immunoprecipitates with PICK1 and PKCα. (C) PSD-95 co-immunoprecipitates with NR2B and stargazin. (D) Stargazin co-immunoprecipitates PSD-95 and GluR2. (E) NR1 (5-nm gold, arrowheads) and GluR2 (15-nm gold) co-localize at superficial dorsal horn synapses. Pre, presynaptic terminal; Post, postsynaptic structure. Scale bar: 100 nm

Article Snippet: The following antibodies were used: rabbit anti-GluR2 (Chemicon, Temecula, CA), mouse anti-GluR2 (Chemicon), rabbit anti-GluR2-p Ser 880 ( Chung et al., 2003 ; Steinberg et al., 2006 ), rabbit ant-PICK1 ( Steinberg et al., 2006 ), rabbit anti-GRIP1 (Upstate, Lake Placid, NY), mouse anti-PSD-95 (Upstate), rabbit anti-PSD-93 (Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2B (Chemicon), rabbit anti-NR1 (Chemicon), rabbit anti-NR2A/2B (Chemicon), rabbit anti-PKCα (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-PKC β (Santa Cruz Biotechnology), rabbit anti-PKCγ (Santa Cruz Biotechnology), rabbit anti- N -cadherin (BD Transduction Laboratories, San Jose, CA), rabbit anti-stargazin (Upstate), mouse anti-α-adaptin (Sigma), and mouse anti-β-actin (Sigma).

Techniques:

Journal:

Article Title: Persistent inflammation induces GluR2 internalization via NMDA receptor-triggered PKC activation in dorsal horn neurons

doi: 10.1523/JNEUROSCI.4514-08.2009

Figure Lengend Snippet: Proposed model for the NMDAR/PKC-dependent dorsal horn GluR2 internalization under persistent inflammatory pain conditions. NMDARs couple to AMPARs through PSD-95 (which binds to NR2A/2B) interaction with stargazin (which binds to GluR1, GluR2, and GluR4). Under normal conditions, ABP/GRIP binds to and anchors GluR2 at synapses. Under persistent inflammatory pain conditions, NMDAR activation causes Ca2+ influx and PKCα activation. The latter phosphorylates GluR2 at Ser880 and disrupts GluR2 binding to ABP/GRIP, which leads to GluR2 internalization. GluR2 internalization results in an increase of AMPAR Ca2+ permeability. The increase in [Ca2+]i in dorsal horn neurons should initiate or potentiate a variety of Ca2+-dependent intracellular cascades that are associated with the maintenance of persistent inflammatory pain.

Article Snippet: The following antibodies were used: rabbit anti-GluR2 (Chemicon, Temecula, CA), mouse anti-GluR2 (Chemicon), rabbit anti-GluR2-p Ser 880 ( Chung et al., 2003 ; Steinberg et al., 2006 ), rabbit ant-PICK1 ( Steinberg et al., 2006 ), rabbit anti-GRIP1 (Upstate, Lake Placid, NY), mouse anti-PSD-95 (Upstate), rabbit anti-PSD-93 (Alomone Labs Ltd, Jerusalem, Israel), rabbit anti-NR2B (Chemicon), rabbit anti-NR1 (Chemicon), rabbit anti-NR2A/2B (Chemicon), rabbit anti-PKCα (Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-PKC β (Santa Cruz Biotechnology), rabbit anti-PKCγ (Santa Cruz Biotechnology), rabbit anti- N -cadherin (BD Transduction Laboratories, San Jose, CA), rabbit anti-stargazin (Upstate), mouse anti-α-adaptin (Sigma), and mouse anti-β-actin (Sigma).

Techniques: Activation Assay, Binding Assay, Permeability

Journal:

Article Title: Chronic administration of docosahexaenoic acid or eicosapentaenoic acid, but not arachidonic acid, alone or in combination with uridine, increases brain phosphatide and synaptic protein levels in gerbils

doi: 10.1016/j.neuroscience.2007.06.016

Figure Lengend Snippet: Effects of AA, DHA or EPA, alone or in combination with a UMP-supplemented diet, on levels of the presynaptic or postsynaptic proteins PSD-95 (a1, a2); Synapsin-1 (b1, b2) and Syntaxin-3 (c1, c2). CV, control diet + vehicle; CA, control diet + AA; CD, control diet + DHA; CE, control diet + EPA; UV, UMP-supplemented diet + vehicle; UA, UMP-supplemented diet + AA; UD, UMP-supplemented diet + DHA; UE, UMP-supplemented diet + EPA. *P<0.05; **P<0.01; and ***P<0.001 compared with CV, and aP<0.05 compared with CA on the left-sided columns (a1, b1, and c1) using One Way ANOVA. *P<0.05; **P<0.01; and ***P<0.001 compared with UV, and xP<0.05; and yP<0.01 compared with UA on the right-sided columns (a2, b2, and c2) using One Way ANOVA.

Article Snippet: Membranes were then rinsed five times in TBST buffer and immersed in TBST solution containing the antibody of interest (rabbit anti-syntaxin-3 [Abcam, Cambridge, MA, USA], mouse anti-PSD-95 [Upstate, Lake Placid, NY, USA], mouse anti-synapsin-1 [Calbiochem, San Diego, CA, USA], and mouse anti-β-tubulin [Chemicon, Temecula, CA, USA]).

Techniques: