Transfer of NMDAR2 cDNAs increases endogenous NMDAR1 protein and induces expression of functional NMDA receptors in PC12 cells.
The pheochromocytoma cell line (PC12) has been used as a model system for the study of regulation of expression of NMDA receptors. PC12 cells express a substantial amount of NMDAR1 subunit (NR1) mRNA, whereas they express only a small amount of NR1 protein. The level of functional NMDA receptor expression is almost negligible. To test the possibility that NMDAR2 subunits (NR2) control expression of functional NMDA receptors as well as NR1 protein, we transferred NR2A-D cDNAs into PC12 cells using adenovirus vectors. Prominent NMDA receptor-mediated currents were recorded in PC12 cells to which NR2A or NR2B cDNA was delivered without NR1 cDNA. The amplitudes of these responses were similar to those in PC12 cells to which NR1 cDNA was delivered together with NR2A or NR2B cDNA. In cells to which either NR2C or NR2D cDNA alone was delivered, NMDA receptor-mediated currents were also detected, although to a much lesser extent. These results showed that NR2 proteins produced by gene transfer are co-assembled with the endogenous NR1 protein to form functional heteromeric receptors. The delivery of NR2A-D cDNAs also increased the amount of NR1 protein but not that of NR1 mRNA, suggesting that this protein increase is due to post-transcriptional mechanisms. The effects of NR2A-B gene transfer on expression of NR1 protein were much more efficient than those of NR2C-D gene transfer.
Saito
Yutaka
Y
Department of Physiology, Gunma University School of Medicine, 3-39-22 Showa-machi, Maebashi 371-8511, Japan. yutakas@med.gunma-u.ac.jp
Tsuzuki
Keisuke
K
Yamada
Nobuaki
N
Okado
Haruo
H
Miwa
Akiko
A
Goto
Fumio
F
Ozawa
Seiji
S
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002490:Central Nervous System / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D018014:Gene Transfer Techniques; D018698:Glutamic Acid / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D008954:Models, Biological; D016716:PC12 Cells; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics* / Q000378:metabolism*; D009435:Synaptic Transmission / Q000235:genetics
D018076:DNA, Complementary; C409466:NR1 NMDA receptor; C120997:NR2A NMDA receptor; C121001:NR2B NMDA receptor; C432823:NR2C NMDA receptor; C464436:NR2D NMDA receptor; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid