The expression of the N-methyl-D-aspartate (NMDA) receptor subunit mRNAs NMDAR1 and NMDAR2A-D was characterized in undifferentiated and nerve growth factor (NGF)-differentiated PC12 cells using Northern blotting, RNase protection assays (RPA) and polymerase chain reaction (PCR). PC12 cells expressed predominately the splice variant NMDAR1-4a and smaller amounts of NMDAR1-1a, NMDAR1-2a and NMDAR1-3a. No splice isoforms containing exon 5 were detected. The NMDAR2C subunit was detected in PC12 cells by Northern blotting and trace amounts of NMDAR2A, B and D were detected by PCR. PC12 cells may be a useful model system for the study of the transcriptional and post-transcriptional regulation of expression of the NMDA receptor subunit genes, including the alternative splicing of NMDAR1 pre-mRNAs.