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Molecular Pharmacology

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Research ArticleArticle

Cycloheximide Increases Proenkephalin and Tyrosine Hydroxylase Gene Expression in Rat Adrenal Medulla

Je-Seong Won, Jin-Koo Lee, Dong-Keun Song, Sung-Oh Huh, Jun-Sub Jung, Yung-Hi Kim, Mi-Ran Choi and Hong-Won Suh
Molecular Pharmacology June 2000, 57 (6) 1173-1181;
Je-Seong Won
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Jin-Koo Lee
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Dong-Keun Song
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Sung-Oh Huh
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Jun-Sub Jung
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Yung-Hi Kim
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Mi-Ran Choi
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Hong-Won Suh
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Abstract

The effect of cycloheximide (CHX; 5 mg/kg) on proenkephalin (proENK) and tyrosine hydroxylase (TH) mRNA expression in rat central and peripheral nervous systems was studied. CHX increased proENK and TH mRNA levels in the adrenal gland, but not in hippocampus, striatum, midbrain, brainstem, pituitary, and hypothalamus. The pretreatment with actinomycin D (0.5 mg/kg) significantly decreased CHX-induced proENK and TH mRNA expression, suggesting that the CHX-dependent increase of these mRNA levels may be caused by the increase of transcriptional activity rather than RNA stabilization. To investigate the factors involved in CHX-induced proENK and TH mRNA expression, the effect of CHX on activator protein-1 (AP-1), cAMP response element (CRE) binding protein (CREB), and glucocorticoid response element (GRE) was tested. In AP-1, the basal expression of Fra-2 and c-Jun proteins and AP-1 DNA binding activity in the adrenal medulla was higher than other tissues tested, but CHX reduced these protein levels and AP-1 DNA binding activity. In CREB, CHX time dependently increased the level of phospho-CREB without altering total CRE level and CRE DNA binding activity. Furthermore, phospho-CREB actively participated in CRE DNA binding activity. In GRE, although CHX increased plasma and adrenal corticosterone level, RU486 (10 mg/kg) reduced CHX-induced proENK, but not TH, mRNA level in a partial manner. These results suggest that the basal expression of proENK and TH mRNA transcription in the adrenal gland seems to be tonically inhibited by de novo protein synthesis. In addition, CHX-dependent increase of proENK and TH mRNA expression in the adrenal medulla is well correlated with phospho-CREB level, but not AP-1. Finally, glucocorticoid seems to be involved at least partially in CHX-dependent proENK, but not TH, mRNA expression in the adrenal medulla.

Footnotes

  • Send reprint requests to: Hong-Won Suh, Ph.D., Department of Pharmacology, College of Medicine, Hallym University, 1 Okchun-Dong, Chunchon, Kangwon-Do, 200-702, Korea. E-mail: hwsuh{at}sun.hallym.ac.kr

  • ↵1 This work was supported by the Research Grant (981-0714-101-2) from Korea Science and Engineering Foundation.

  • Abbreviations:
    ME
    [Met5]enkephalin
    proENK
    proenkephalin
    TH
    tyrosine hydroxylase
    AP-1
    activator protein-1
    CRE
    cAMP response element
    CREB
    CRE-binding protein
    GRE
    glucocorticoid response element
    CHX
    cycloheximide
    p38
    p38 mitogen-activated kinase
    JNK
    c-Jun NH2-terminal kinase, ERK, extracellular signal response kinase
    SSC
    standard saline citrate
    DIG
    digoxigenin
    TBS
    Tris-buffered saline
    AMD
    actinomycin D
    ATF-2
    activating transcription factor-2
    MAPK
    mitogen-activated protein kinase
    • Received September 14, 1999.
    • Accepted February 3, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 57 (6)
Molecular Pharmacology
Vol. 57, Issue 6
1 Jun 2000
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Research ArticleArticle

Cycloheximide Increases Proenkephalin and Tyrosine Hydroxylase Gene Expression in Rat Adrenal Medulla

Je-Seong Won, Jin-Koo Lee, Dong-Keun Song, Sung-Oh Huh, Jun-Sub Jung, Yung-Hi Kim, Mi-Ran Choi and Hong-Won Suh
Molecular Pharmacology June 1, 2000, 57 (6) 1173-1181;

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Research ArticleArticle

Cycloheximide Increases Proenkephalin and Tyrosine Hydroxylase Gene Expression in Rat Adrenal Medulla

Je-Seong Won, Jin-Koo Lee, Dong-Keun Song, Sung-Oh Huh, Jun-Sub Jung, Yung-Hi Kim, Mi-Ran Choi and Hong-Won Suh
Molecular Pharmacology June 1, 2000, 57 (6) 1173-1181;
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