Immunoprecipitation, Immunoblotting, and Lmmunocytochemistry Studies Suggest That Glutamate Receptor 6 Subunits Form Novel Postsynaptic Receptor Complexes
The Journal of Neuroscience, March 1995, 75(3): 2533-2546 Immunoprecipitation, Immunoblotting, and lmmunocytochemistry Studies Suggest That Glutamate Receptor 6 Subunits Form Novel Postsynaptic Receptor Complexes Ebrahim Mayat, Ronald S. Petralia, Ya-Xian Wang, and Robert J. Wenthold Laboratory of Neurochemistry, NIDCD, NIH, Bethesda, Maryland 20892 An antibody was made to a C-terminus peptide of the glu- specific ion channels (for a recent review, see Hollmann and tamate receptor 62 subunit and used to study the distri- Heinemann, 1994). According to pharmacological and subse- bution, biochemical properties, and developmental expres- quent binding studies, GluRs were classified into three main sion of the S receptor in rat brain. The antibody recognizes subtypes: the NMDA, a-amino-3-hydroxy-5-methyl-4-isoxazole both 61 and 62 but not AMPA, kainate, NMDA, and propionic acid (AMPA), and kainate (KA) receptors (for mGluRla glutamate receptor subunits based on Western reviews, seeMonaghan et al., 1989; Watkins et al., 1990; Young blot analysis of transfected HEK-293 cells. Western blot and Fagg, 1990). In the last 5 years, cloning studieshave led to analysis of brain showed a single immunoreactive band, the characterization of multiple subunits which comprise each migrating at M, = 114,000. lmmunoprecipitation of deter- of the three GluR subtypes (for reviews, see Nakanishi, 1992; gent-solubilized cerebellar membranes was done to deter- Sommer and Seeburg, 1992; Hollmann and Heinemann, 1994). mine if S is associated with other glutamate receptor sub- The AMPA receptor has four subunits,GluRl-4, or GluR-A to units and if it binds any of the common excitatory amino -D (Hollmann et al., 1989; Boulter et al., 1990; Keinanen et al, acid ligands.
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