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Cloning and Localization of Exon 5‐Containing Isoforms of the NMDAR1 Subunit in Human and Rat Brains
Author(s) -
Nash Norman R.,
Heilman Craig J.,
Rees Howard D.,
Levey Allan I.
Publication year - 1997
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1997.69020485.x
Subject(s) - gene isoform , exon , microbiology and biotechnology , splice , cloning (programming) , protein subunit , alternative splicing , biology , monoclonal antibody , epitope , western blot , fusion protein , receptor , biochemistry , antibody , gene , genetics , recombinant dna , computer science , programming language
Nine isoforms of the rat NMDAR1 receptor subunit have been previously identified, of which several have an alternatively spliced N‐terminal insert believed to be important in proton sensitivity of the receptor. The cloning of the human homologues of NMDAR1‐3b (hNMDA1‐1) and NMDAR1‐4b (hNMDA1‐2), both bearing the insert, is reported here. A monoclonal antibody generated against the N‐terminal region of these isoforms showed reactivity with at least two distinct human brain proteins of ∼115 kDa. This antibody was further characterized by using a series of truncated fusion proteins and splice variants of NMDAR1 demonstrating its specific recognition of an epitope within the 21‐amino acid N‐terminal insert, encoded by exon 5. Western blot and immunocytochemical studies were performed to examine the expression of the exon 5‐containing isoforms of the NMDAR1 subunit in both rat and human brain.