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Molecular Pharmacology

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Research ArticleArticle

Neuron-Restrictive Silencer Factor Regulates the N-Methyl-d-aspartate Receptor 2B Subunit Gene in Basal and Ethanol-Induced Gene Expression in Fetal Cortical Neurons

Mei Qiang, C. S. Sheela Rani and Maharaj K. Ticku
Molecular Pharmacology June 2005, 67 (6) 2115-2125; DOI: https://doi.org/10.1124/mol.104.010751
Mei Qiang
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C. S. Sheela Rani
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Maharaj K. Ticku
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Abstract

Neuron-restrictive silencer factor (NRSF) is a transcriptional repressor of multiple neuronal genes. This study addressed the role of NRSF in N-methyl-d-aspartate (NMDA) receptor NR2B promoter activity and the molecular mechanisms of ethanol-induced NR2B up-regulation in fetal cortical neurons. The 5′-flanking region of the NR2B gene contains five NRSE-like elements. Functional analysis of the upstream regions of the NR2B gene by transient transfection of neurons revealed that neuron-restrictive silencer element (NRSE) motifs located between base pair –1407 and –2741 represses transcription of the gene. Analysis by electrophoretic mobility shift assay and reporter gene assay identified NRSE2 and 3 as responsible for repressing NR2B gene transcription. The identity of NRSF as the functional binding factor is suggested by the specific binding of in vitro synthesized NRSF or cell lysate to the labeled probes and the specific antibody-induced supershift. Furthermore, whereas mutations of NRSE2 and 3 motifs increased the promoter activity, overexpression of NRSF reduced it significantly. The pattern of NRSF expression during development was investigated and demonstrated that the highest expression is on embryonic day 14 with moderate expression on postnatal day 0, reflecting a possible role of NRSF as a regulator during development. Treatment of cultured cortical neurons with 100 mM ethanol for 5 days caused a significant decrease in the NRSF mRNA and protein levels, NRSF/NRSE binding activity, and an increase in the promoter activity. Therefore, our studies suggest that NRSF is a negative regulator of NR2B expression and may contribute to the ethanol-induced up-regulation of the NR2B gene in fetal cortical neurons.

  • Received December 22, 2004.
  • Accepted March 8, 2005.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 67 (6)
Molecular Pharmacology
Vol. 67, Issue 6
1 Jun 2005
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Research ArticleArticle

Neuron-Restrictive Silencer Factor Regulates the N-Methyl-d-aspartate Receptor 2B Subunit Gene in Basal and Ethanol-Induced Gene Expression in Fetal Cortical Neurons

Mei Qiang, C. S. Sheela Rani and Maharaj K. Ticku
Molecular Pharmacology June 1, 2005, 67 (6) 2115-2125; DOI: https://doi.org/10.1124/mol.104.010751

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Research ArticleArticle

Neuron-Restrictive Silencer Factor Regulates the N-Methyl-d-aspartate Receptor 2B Subunit Gene in Basal and Ethanol-Induced Gene Expression in Fetal Cortical Neurons

Mei Qiang, C. S. Sheela Rani and Maharaj K. Ticku
Molecular Pharmacology June 1, 2005, 67 (6) 2115-2125; DOI: https://doi.org/10.1124/mol.104.010751
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