Journal: The Journal of Neuroscience
Article Title: A Specific Role for NR2A-Containing NMDA Receptors in the Maintenance of Parvalbumin and GAD67 Immunoreactivity in Cultured Interneurons
Figure Lengend Snippet: Cultured cortical interneurons represent a nonoverlapping population with respect to the expression of calcium-binding proteins. Dissociated cortical neurons were grown for 21 d in vitro and fixed in 4% paraformaldehyde. Double immunocytochemistry was performed using a rabbit polyclonal antibody against PV (red) and a mouse monoclonal antibody (green) against calretinin (top), calbindin (middle), or αCaMKII (bottom) and AlexaFluor-conjugated secondary antibodies. Scale bar, 15 μm.
Article Snippet: For double immunostaining, the coverslips were incubated in 2% normal goat serum containing a dilution of either a mouse monoclonal antibody against parvalbumin (1:4000; Sigma, St. Louis, MO) or a dilution of a rabbit polyclonal antibody against the same protein (1:7000; Swant, Bellinzona, Switzerland), and a dilution of a rabbit polyclonal or mouse monoclonal antibodies against α calcium calmodulin-dependent protein kinase II (αCaMKII) (ABR), against calbindin or calretinin (Sigma), against GAD67, glutamate receptor 1 (GluR1), GluR2,3, metabotropic GluR1 (mGluR1), or mGluR5 (Chemicon, Temecula, CA) or against NR2A and NR2B (Invitrogen; Upstate Biotechnology, Lake Placid, NY; Santa Cruz Biotechnology, Santa Cruz, CA) for 1–2 h at 37°C.
Techniques: Cell Culture, Expressing, Binding Assay, In Vitro, Immunocytochemistry