Stable transfection of the NR1 subunit in Chinese hamster ovary cells fails to produce a functional N-methyl-D-aspartate receptor.
The NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor cDNA was stably transfected into Chinese hamster ovary (CHO) cells. Northern analysis revealed 3 clonal cell lines expressing high levels of NMDAR1 mRNA (BA1, BA2 and BA3). NMDAR1 protein was readily detected by Western analysis in only one of the clonal populations, BA1. Whole-cell patch clamp analysis revealed a lack of functional NMDA receptors in all of the cell lines, including BA1. NMDA-mediated responses were obtained only after a second subunit, NMDAR2A, was transiently expressed in BA1 cells. Therefore, the NR1 subunit alone may not be sufficient to form functional NMDA receptors in CHO cells.
Boeckman
F A
FA
Department of Neurobiology, University of Pittsburgh School of Medicine, PA 15261.
Aizenman
E
E
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000096:biosynthesis; D004594:Electrophysiology; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D016194:Receptors, N-Methyl-D-Aspartate / Q000096:biosynthesis* / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D016194:Receptors, N-Methyl-D-Aspartate