Cloning and localization of exon 5-containing isoforms of the NMDAR1 subunit in human and rat brains.
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 approximately 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.
Nash
N R
NR
Department of Neurology, Emory University School of Medicine, Atlanta, Georgia 30322, U.S.A.
Heilman
C J
CJ
Rees
H D
HD
Levey
A I
AI
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015153:Blotting, Western; D001923:Brain Chemistry; D003001:Cloning, Molecular; D018604:Epitope Mapping; D005091:Exons; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D017385:Sequence Homology
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D011993:Recombinant Fusion Proteins