Journal: The Journal of Biological Chemistry
Article Title: Activation of Intracellular Metabotropic Glutamate Receptor 5 in Striatal Neurons Leads to Up-regulation of Genes Associated with Sustained Synaptic Transmission Including Arc/Arg3.1 Protein *
doi: 10.1074/jbc.M111.301366
Figure Lengend Snippet: Intracellular mGluR5 plays a critical role in glutamate and neuronal activity-induced Arc. A, striatal neurons were pretreated with NMDA antagonist MK801 (5 μm), the AMPA/kainate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 μm), and mGluR1 antagonist CPCCOEt (20 μm) followed by treatment with glutamate (100 μm, 1 h). Glutamate increased Arc expression that was blocked by MPEP but not LY393053. Bars indicate Arc-positive/total neurons compared with control. #, p < 0.05 versus control; n = 4, more than 2500 neurons assessed per treatment. Ctl, control. B, bath application of 15 mm KCl-induced Arc in ∼30% striatal neurons. This KCl-regulated induction was decreased ∼28% by MPEP, whereas LY395053 + KCl was not significantly different from KCl alone. Bars indicate Arc positive/total neurons expressed as the percentage of KCl treatment. More than 2000 neurons per treatment from four independent experiments were assessed for Arc immunofluorescence using the High Content Imager. #, p < 0.0001 compared with untreated control. *, p < 0.005 compared with KCl treated alone. C, the proposed model for intracellular mGluR5-mediated transcriptional activation of Arc/Arg3.1 is shown. NPC, nuclear pore complex.
Article Snippet: Primary antibodies included polyclonal anti-C-terminal mGluR5 (1:250; Millipore), mouse monoclonal C-7 anti-Arc/Arg3.1 (1:100; Santa Cruz Biotechnology), rabbit polyclonal anti-SRF (1:1000, Santa Cruz), anti-NeuN (1:100, Millipore), monoclonal anti-lamin B 2 (1:100; Zymed Laboratories Inc.), polyclonal anti-EAAT3 (1:1000; Chemicon International, Temecula, CA), polyclonal anti-quisqualate antibody (1:250; Dr. J. Koerner, University of Minnesota), and polyclonal anti-pERK1/2 (1:200; Cell Signaling Technology, Beverly, MA).
Techniques: Activity Assay, Expressing, Immunofluorescence, Activation Assay
Santa Cruz Biotechnology is a verified supplier 
