( ts were used for RT-PCR reac...)Gene-Specific Primers for NMDA Receptor Subunits

ts were used for RT-PCR reactions to identify cell populations expressing these specific NMDA receptor subunit genes. Briefly, RT-PCR was conducted using species-specific primer sets for various NMDA receptor subunits (table 1) with cDNA constructed from whole rat brain (figure 1). (Data from reactions using human brain and human hNT cells as templates are not shown). PCR products resulting from amplification of rat NMDA receptor subunit genes were electrophoresed on an agarose gel (figure 1), and their sizes were visualized by ethidium bromide staining. Amplification products with the gene-specific primers for the NMDA receptor subunits resulted in products with correct sizes for all cDNAs tested. For NR1, amplification using primers flanking the C1 or N1 splice-variant exons from rat resulted in two products of different sizes, corresponding to mRNAs either containing or lacking these exons.

Conclusions

Investigators can now use the extremely sensitive RT-PCR method to study expression of NMDA receptor subunit genes. Stratagenes RT-PCR primers for rat and human NMDA receptor genes have been tested using cDNA templates derived from whole brain or cultured hNT cells. These new RT-PCR primers will allow further studies of the subunit composition of NMDA receptors.

REFERENCES

1. Collingridge, G.L., et al. (1983) J. Physiol. (Lond.) 335: 33-46.

2. Moriyoshi, K., et al. (1991) Nature 354: 31-37.

3. Monyer, H., et al. (1992) Science 256: 1217-1221.

4. Laurie, D.J., Seeburg, P.H. (1994) J. Neurosci. 14: 3180-3194.

5. Monyer, H., et al. (1994) Neuron 12: 529-540.

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