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The Journal of Neuroscience, September 15, 1999, 19(18):7781-7792
Alternative Splicing of the C-Terminal Domain Regulates Cell Surface Expression of the NMDA Receptor NR1 Subunit
Shigeo Okabe1, 2, Akiko Miwa3, and Haruo Okado3 1 Laboratory of Molecular Neurobiology, National Institute of Bioscience and Human Technology, Tsukuba, Ibaraki 305-8566, Japan, 2 Department of Anatomy and Cell Biology, School of Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, 113-8519, Japan, and 3 Department of Neurobiology, Tokyo Metropolitan Institute for Neuroscience, Fuchu, Tokyo 183-8526, Japan
Subcellular localization of the NMDA receptor NR1 splice forms was studied by expressing individual splice variants and their epitope-tagged derivatives in mouse fibroblasts and in hippocampal neurons. When NR1 splice variants were expressed in fibroblasts, the amount of NR1 molecules expressed on the cell surface varied among forms with different C-terminal cytoplasmic domains. The splice forms with the longest C-terminal cytoplasmic tail (NR1-1a and NR1-1b) showed the lowest amount of cell surface expression, and the splice forms with the shortest C-terminal cytoplasmic tail (NR1-4a and NR1-4b) showed the highest cell surface expression. Cell surface expression of NR1 was enhanced by the coexpression of the NR2 subunit. We measured the glutamate-induced increase of calcium concentration in fibroblasts expressing one of the NR1 splice forms and the NR2B subunit. The increase of calcium concentration after glutamate application had a positive correlation with the amount of NR1 splice forms expressed on the cell surface. When epitope-tagged NR1 splice variants were expressed in primary hippocampal neurons using recombinant adenoviruses, we also observed the differential expression on the cell surface between splice variants. These results suggest that the splicing of the C-terminal domain of the NR1 subunit regulates the cell surface expression of the functional NMDA receptors.
Key words: NMDA receptor; alternative splicing; fluorescent antibody technique; calcium imaging; membrane proteins; neurons
Copyright © 1999 Society for Neuroscience 0270-6474/99/19187781-12$05.00/0
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