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Molecular Pharmacology

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Research ArticleArticle

Expressional Down-Regulation of Neuronal-Type Nitric Oxide Synthase I by Glucocorticoids in N1E-115 Neuroblastoma Cells

Petra M. Schwarz, Birgit Gierten, Jean-Paul Boissel and Ulrich Förstermann
Molecular Pharmacology August 1998, 54 (2) 258-263; DOI: https://doi.org/10.1124/mol.54.2.258
Petra M. Schwarz
Department of Pharmacology, Johannes Gutenberg University, 55101 Mainz, Germany
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Birgit Gierten
Department of Pharmacology, Johannes Gutenberg University, 55101 Mainz, Germany
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Jean-Paul Boissel
Department of Pharmacology, Johannes Gutenberg University, 55101 Mainz, Germany
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Ulrich Förstermann
Department of Pharmacology, Johannes Gutenberg University, 55101 Mainz, Germany
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Abstract

Neuronal-type nitric oxide synthase (NOS I) is involved in ischemia-induced brain damage, and glucocorticoids have been reported to protect from brain damage. This prompted us to investigate if the activity or expression of NOS I was influenced by glucocorticoids. We used the murine neuroblastoma cell line N1E-115 as our experimental model. Short-term incubation (30 min) of the N1E-115 cells with dexamethasone (10 nm to 1 μm) or hydrocortisone (100 nm to 10 μm) did not change the enzymatic activity of NOS I. However, the glucocorticoids inhibited NOS I mRNA expression in a concentration-dependent fashion (down to 53.3 ± 2.5% of control). In time-course experiments with 100 nm dexamethasone, maximum down-regulation of NOS I mRNA was seen after 24 hr (55.6 ± 6.3% of control). Similar effects were seen with 10 μm hydrocortisone. The effect of 100 nm dexamethasone was completely reversed by 1 μm of the glucocorticoid receptor antagonist mifepristone. In experiments with actinomycin D (10 μg/ml), the half-life of the NOS I mRNA was determined to be approximately 12 hr and remained unchanged after glucocorticoid incubation. Nuclear run-on analyses indicated that the decrease in NOS I mRNA was the result of a glucocorticoid-induced inhibition of NOS I gene transcription. In Western blots, the 160-kDa NOS I protein band was down-regulated to 68.5 ± 8.4% of control after an incubation of the N1E-115 cells with 100 nm dexamethasone for 26 hr. Similarly, NO production was down-regulated to 57.8 ± 8.7% of control. These data demonstrate that glucocorticoids reduce the expression of NOS I without changing its activity.

Footnotes

    • Received January 22, 1998.
    • Accepted May 5, 1998.
  • Send reprint requests to: Dr. Petra M. Schwarz, Department of Pharmacology, Johannes Gutenberg University, Obere Zahlbacher Strasse 67, 55101 Mainz, Germany. E-mail:petra.schwarz{at}uni-mainz.de

  • This work was supported by Grants Fo 144/3–2 and SFB 553 (Project A1) from the Deutsche Forschungsgemeinschaft (Bonn, Germany) and by a grant from the Ministry of the Environment of the State of Rhineland-Palatinate (Mainz, Germany). This article is part of the thesis work of B.G.

  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 54 (2)
Molecular Pharmacology
Vol. 54, Issue 2
1 Aug 1998
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Research ArticleArticle

Expressional Down-Regulation of Neuronal-Type Nitric Oxide Synthase I by Glucocorticoids in N1E-115 Neuroblastoma Cells

Petra M. Schwarz, Birgit Gierten, Jean-Paul Boissel and Ulrich Förstermann
Molecular Pharmacology August 1, 1998, 54 (2) 258-263; DOI: https://doi.org/10.1124/mol.54.2.258

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Research ArticleArticle

Expressional Down-Regulation of Neuronal-Type Nitric Oxide Synthase I by Glucocorticoids in N1E-115 Neuroblastoma Cells

Petra M. Schwarz, Birgit Gierten, Jean-Paul Boissel and Ulrich Förstermann
Molecular Pharmacology August 1, 1998, 54 (2) 258-263; DOI: https://doi.org/10.1124/mol.54.2.258
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