Agonist and cyclic AMP-mediated regulation of beta 1-adrenergic receptor mRNA and gene transcription in rat C6 glioma cells

J Neurochem. 1994 Nov;63(5):1635-45. doi: 10.1046/j.1471-4159.1994.63051635.x.

Abstract

Exposure of rat C6 glioma cells to either agonists or agents that increase cyclic AMP levels leads to down-regulation of beta 1-adrenergic receptors (beta 1 AR) as measured by loss of radioligand binding sites. The present study examines the influence of isoproterenol and forskolin treatment on levels of beta 1 AR mRNA, mRNA stability, and gene transcription rate. Isoproterenol treatment of C6 cells altered beta 1 AR mRNA levels in a biphasic manner; i.e., short-term exposure (30-60 min) increased by 50%, whereas longer exposure (2-6 h) decreased by 50% the levels of beta 1 AR mRNA. The extent of both the up- and down-regulation was dependent on agonist concentration. Similar regulation of beta 1 AR mRNA was observed in forskolin-treated cells. Pretreatment of the cells with Pseudomonas exotoxin A, a potent inhibitor of protein synthesis, completely blocked isoproterenol- and forskolin-mediated down-regulation of beta 1 AR mRNA, and thereby potentiated the increase in receptor mRNA up to fourfold over the 6-h time course. The mechanisms underlying beta 1 AR mRNA down-regulation were examined. The half-life of beta 1 AR mRNA was slightly increased (from 61 to 77 min) after a 2-h exposure of the cells to either isoproterenol or forskolin. Nuclear run-on analysis demonstrated that the rate of beta 1 AR gene transcription was increased after isoproterenol incubation for 60 min, but then decreased after 90-240 min, consistent with the time course for up- and down-regulation of beta 1 AR mRNA. Isoproterenol treatment (120 min) also decreased the level of beta 1 AR nascent transcripts, purified by affinity chromatography of RNA isolated from 4-thiouridine-pulsed cells. The results demonstrate that beta 1 AR mRNA has a relatively short half-life and that agonist regulation of beta 1 AR mRNA is mediated by activation of the cyclic AMP system. Moreover, the results indicate that agonist regulation of beta 1 AR mRNA occurs at the level of beta 1 AR gene transcription, not mRNA stability. Finally, the observed requirement for protein synthesis indicates that beta 1 AR mRNA down-regulation may be mediated by the induction of a repressor of the beta 1 AR gene.

Publication types

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

MeSH terms

  • Adrenergic beta-1 Receptor Agonists*
  • Animals
  • Colforsin / pharmacology*
  • Cyclic AMP / agonists*
  • Cyclic AMP / pharmacology*
  • Cyclic AMP / physiology
  • Down-Regulation
  • Endotoxins / pharmacology
  • Glioma / genetics*
  • Glioma / metabolism
  • Glioma / pathology*
  • Isoproterenol / pharmacology*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Rats
  • Receptors, Adrenergic, beta-1 / genetics*
  • Receptors, Adrenergic, beta-1 / physiology
  • Time Factors
  • Transcription, Genetic*
  • Tumor Cells, Cultured

Substances

  • Adrenergic beta-1 Receptor Agonists
  • Endotoxins
  • RNA, Messenger
  • Receptors, Adrenergic, beta-1
  • Colforsin
  • Cyclic AMP
  • Isoproterenol