Navigation Links
Gene-Specific Primers for NMDA Receptor Subunits

Additional tools for neuroscience researchers

Robert Buchner Peter L. Pingerelli Michael D. Browning Mary Ellen Simcox
Stratagene Cloning Systems, Inc.

Stratagene is now offering gene-specific primer sets for the rat and human NMDA receptor. In this article, we describe using these primers in reverse transcriptase-mediated polymerase chain reaction (RT-PCR) assays to show expression of NMDA subunits with Stratagenes rat cDNA, human cDNA and hNT neuron cDNA libraries as templates. By using these primers, researchers will be able to study rat and human NMDAR subunit-specific expression in a variety of different cell types and regions of the brain.

The ion channels activated by glutamate are typically divided into two classes. The ion channels that are sensitive to N-methyl-D-aspartate (NMDA) are designated NMDA receptors (NMDAR), while those activated by kainate and a-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA) are known as kainate/AMPA receptors (K/AMPAR). The NMDAR assumes a critical role in long-term potentiation (LTP), a putative cellular substrate of memory.1 This receptor has also been linked to neuronal development and has been implicated in several disorders of the central nervous system, including epilepsy and ischemic neuronal cell death.

The rat NMDAR1 (NR1) was the first subunit of the NMDAR to be cloned.2 The NR1 protein can form NMDA-activated ion channels when expressed in Xenopus oocytes, but the currents in such channels are much weaker than those seen in situ. Channels with more relevant physiological characteristics are produced when the NR1 subunit is combined with one or more of the NMDAR2 (NR2A-D) subunits.3 In addition, there are a number of different splice variants of the NR1.4 Significant functional diversity has be en seen in recombinant systems with heterologous coexpression of various NR2 subunits and/or NR1 splice variants.5 However, little is known about the expression of the NMDAR in situ.

NMDA Receptor Subunit

Expected PCR Product Size (bp)

NR1, Rat; flanking C1 exon

C1+, 214 C1, 103

NR1, Rat: flanking N1 exon

N1+, 235 N1,172

NR2A, Rat


NR2B, Rat


NR1, Human


Table 1

figure 1

RT-PCR assays with gene-specific primer sets can be used to study gene expression of the NMDA receptor subunits.6 Stratagenes cDNA library templates and amplification primer sets6 for rat NMDA subuni 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.


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.

  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.

  6. Sheng, M., et al. (1994) Nature 368: 144-147.



Page: All 1 2 3 4

Related biology technology :

1. Control RT-PCR Primers for Human Gene Transcripts with Varying Abundance
2. RT-PCR Primers for the Study of Apoptosis
3. New PCR Primers for Human Diseases
4. Amplification of Epstein-Barr Virus Exon C with Difficult Primers Using Eppendorf Mastercycler gradient
5. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
6. Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
7. pFB-ERV: Retroviral Delivery of the Ecdysone Receptor Proteins
8. TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins
9. Quantification of the Purinergic Receptor P2X3 using ICAT and Orthogonal 2D LC-MS/MS with an Ion Trap Mass Spectrometer
10. Isolation of Low Molecular Weight Digestion Products of the Human Platelet Thromboxane A2 Receptor, Rev A
11. Performance of Ad-A-Gene EGFP-Glucocorticoid Receptor (EGFP-GCCR): an adenoviral vector gene delivery system
Post Your Comments:

(Date:6/27/2016)... 2016  Global demand for enzymes is forecast ... to $7.2 billion.  This market includes enzymes used ... biofuel production, animal feed, and other markets) and ... Food and beverages will remain the largest market ... of products containing enzymes in developing regions.  These ...
(Date:6/27/2016)... ... June 27, 2016 , ... Newly created 4Sight ... solutions to the healthcare market. The company's primary focus is on new product ... marketing strategies that are necessary to help companies efficiently bring their products to ...
(Date:6/24/2016)... SAN DIEGO , June 24, 2016 /PRNewswire/ ... that more sensitively detects cancers susceptible to PARP ... individual circulating tumor cells (CTCs). The new test ... of HRD-targeted therapeutics in multiple cancer types. ... therapies targeting DNA damage response pathways, including PARP, ...
(Date:6/23/2016)... Mass. , June 23, 2016   ... development of novel compounds designed to target cancer ... napabucasin, has been granted Orphan Drug Designation from ... the treatment of gastric cancer, including gastroesophageal junction ... stemness inhibitor designed to inhibit cancer stemness pathways ...
Breaking Biology Technology:
(Date:6/2/2016)... NEW YORK , June 2, 2016   The ... (Weather), is announcing Watson Ads, an industry-first capability in which ... advertising, by being able to ask questions via voice or ... Marketers have long ... with the consumer, that can be personal, relevant and valuable; ...
(Date:6/1/2016)... YORK , June 1, 2016 ... Technology in Election Administration and Criminal Identification to Boost ... to a recently released TechSci Research report, " Global ... By Region, Competition Forecast and Opportunities, 2011 - 2021", ... 24.8 billion by 2021, on account of growing security ...
(Date:5/20/2016)... , May 20, 2016  VoiceIt is excited ... with VoicePass. By working together, VoiceIt ...  Because VoiceIt and VoicePass take slightly different approaches ... increases both security and usability. ... about this new partnership. "This marketing ...
Breaking Biology News(10 mins):