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Papers In Press, published online ahead of print September 27, 2002
J. Biol. Chem, 10.1074/jbc.M203032200
Submitted on March 28, 2002
Revised on September 25, 2002
Accepted on September 27, 2002
LDN, NIH/NICHD, Bethesda, Maryland 20892
Corresponding Author: buonanno{at}helix.nih.gov
SUMMARY The postnatal appearance and upregulation of the NR2A subunit of the NMDA receptor contributes to the functional heterogeneity of the receptor during development. To elucidate the molecular mechanisms that regulate the neural- and developmental-specific expression of NR2A, an upstream ~9kb region of the gene harboring the promoter was isolated and characterized in transgenic mice and transfected cortical neurons. Transgenic mouse lines generated with luciferase reporter constructs driven by either 9kb or 1kb of upstream sequence selectively transcribe the transgene in brain, as compared to other non-neural tissues. Reporter luciferase levels in dissociated cultures made from these mice are over 100-fold greater in neuronal/glial co-cultures than in pure glial cultures. Analysis of NR2A 5’-nested deletions in transfected cultures of cortical neurons and glia indicate that, while sequences residing upstream of -1079bp augment NR2A neuronal expression, sequences between -486bp and -447bp are sufficient to maintain neuronal preference. An RE1/NRSE element is not necessary for NR2A neuron-specificity. Furthermore, comparison of the 5’-deletion constructs in cortical neurons grown for 5, 8, 11 or 14 DIV indicate that sequences between -1253bp and -1180bp are necessary for maturational upregulation of NR2A. Thus, different cis-acting sequences control the regional and temporal expression of NR2A, implicating distinct regulatory pathways.
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