Ethanol increases tyrosine hydroxylase gene expression in N1E-115 neuroblastoma cells

J Biol Chem. 1991 Nov 25;266(33):22279-84.


Catecholamines appear to be involved in behavioral responses to acute and chronic ethanol consumption. Since tyrosine hydroxylase (TH) is the rate-limiting enzyme for catecholamine biosynthesis and is regulated by second messenger systems known to be modulated by ethanol, we studied ethanol-induced changes in TH gene expression. In the N1E-115 neural cell line, Northern and Western blot analyses showed that treatment with 25-200 mM ethanol for 3 days caused a dose-dependent increase in TH mRNA and protein levels. N1E-115 cells were also stably transfected with pTH5'CAT, a plasmid containing 773 base pairs of the TH promoter fused to a chloramphenicol acetyltransferase (CAT) reporter gene. Subclones expressing pTH5'CAT showed ethanol-induced increases in CAT activity, suggesting that ethanol modulates TH gene transcription. Furthermore, simultaneous treatment of transfected cells with 100 mM ethanol and 1 nM to 1 microM prostaglandin E1 increased prostaglandin E1-mediated stimulation of TH-promoter activity. Similarly, simultaneous treatment of transfected cells with 100 mM ethanol and either 10 mM (-)-N6-(R-phenylisopropyl)adenosine or 0.5 mM 8-bromo-cAMP also resulted in increased TH-promoter activity compared to treatment with these agents without ethanol. These results suggest that ethanol treatment of N1E-115 cells has a prominent effect on both basal and cAMP-regulated TH expression. Ethanol-induced changes in TH expression may be a critical molecular event in adaptation of the central nervous system to ethanol.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 8-Bromo Cyclic Adenosine Monophosphate / pharmacology
  • Alprostadil / pharmacology
  • Animals
  • Blotting, Northern
  • Blotting, Western
  • Cell Line
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Ethanol / pharmacology*
  • Gene Expression Regulation, Enzymologic / drug effects*
  • Kinetics
  • Neuroblastoma
  • Phenylisopropyladenosine / pharmacology
  • Plasmids
  • Promoter Regions, Genetic / drug effects*
  • RNA, Messenger / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transfection
  • Tyrosine 3-Monooxygenase / genetics*


  • RNA, Messenger
  • Recombinant Fusion Proteins
  • 8-Bromo Cyclic Adenosine Monophosphate
  • Phenylisopropyladenosine
  • Ethanol
  • Tyrosine 3-Monooxygenase
  • Chloramphenicol O-Acetyltransferase
  • Alprostadil