Quantitation of NMDA receptor NR2 mRNA transcripts in human brain by competitive RT-PCR
| Autor: | Matthew R. Hynd, Peter R. Dodd, H. L. Scott |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Protein subunit
Glutamic Acid Biology Hippocampus Receptors N-Methyl-D-Aspartate Synaptic Transmission Gene expression Humans Protein Isoforms Homomeric RNA Messenger Cloning Molecular Receptor Brain Chemistry Neurons Messenger RNA Reverse Transcriptase Polymerase Chain Reaction General Neuroscience Motor Cortex Brain RNA Templates Genetic Molecular biology Alternative Splicing Synapses Synaptic plasticity NMDA receptor |
| Zdroj: | Brain Research Protocols. 11:67-79 |
| ISSN: | 1385-299X |
| Popis: | The NMDA-selective ionotropic receptor constitutes one of the three principal classes of L-glutamate receptors within the mammalian brain. It plays key roles in neuronal differentiation and synapse consolidation, activity-dependent forms of synaptic plasticity, and excitatory amino acid-mediated neuronal toxicity [Lab. Invest., 68 (1993) 372-387]. NMDA receptors exist as multimeric complexes comprising proteins from two families, NR1 and NR2(A-D) [J. Biol. Chem., 271 (1996) 15669-15674]. Studies on recombinant receptors have revealed that while homomeric NR2 receptors are non-functional, co-expression of an NR1 with an NR2 subunit modulates the efficacy of the resulting channel [Nature, 357 (1992) 70-74]. The RT-PCR assay we describe here was developed to allow quantitation of all hNR2 transcripts in a single-tube PCR assay. Each hNR2 isoform is quantified on the basis of standard curves in which a known amount of synthetic ribonucleic acid competitor is co-amplified against total RNA. The protocol has been applied to the quantitation of hNR2 mRNA levels in autopsy brain. Used in conjunction with a method for the quantitation of hNR1 transcripts [Brain Res. Protoc.. in press]. a complete analysis of NMDA receptor mRNA expression can be obtained. (C) 2003 Elsevier Science B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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