Serum-induced sensitization of cyclic AMP accumulation in C62B rat glioma cells

Myron Lee Toews, J. M. Hoffman, S. A. Liewer, L. J. Arneson-Rotert

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Exposure of C62B rat glioma cells to fresh medium containing fetal bovine serum induced a sensitization of the subsequent ability of isoproterenol and forskolin to stimulate cyclic AMP accumulation, compared to cells exposed to fresh medium without serum. Isoproterenol stimulation was typically increased by 2- to 4-fold and forskolin stimulation by 3- to 5-fold. Sensitization occurred rapidly, was rapidly reversible and appeared to result from an increase in maximal stimulation. A commercial preparation of albumin, purified chromatographically so as to retain bound lipids and other factors, was able to mimic the effect of serum. In contrast to the effects of serum, exposure of cells to phorbol 12-myristate, 13-acetate induced little or no change in forskolin stimulation but a marked desensitization of isoproterenol stimulation that was due primarily to a decrease in potency. Neither the protein kinase C inhibitor staurosporine or overnight exposure to phorbol 12- myristate, 13-acetate to down-regulate protein kinase C prevented serum- induced sensitization. Pertussis toxin almost completely blocked serum- induced sensitization, suggesting involvement of a pertussis toxin-sensitive guanine nucleotide-binding protein in mediating the effects of serum. Sensitization was poorly retained in membrane adenylate cyclase assays. Studies with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, direct assays of cyclic AMP degradation by intact cells and assays of phosphodiesterase activity in cell lysates all indicated that degradation of cyclic AMP was decreased in serum-pretreated cells. Thus, both increased cyclic AMP synthesis and decreased cyclic AMP degradation may contribute to sensitization in these cells.

Original languageEnglish (US)
Pages (from-to)471-478
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume262
Issue number2
StatePublished - Jan 1 1992

Fingerprint

Glioma
Cyclic AMP
Serum
Colforsin
Isoproterenol
Pertussis Toxin
Protein Kinase C
Acetates
1-Methyl-3-isobutylxanthine
Phosphodiesterase Inhibitors
Guanine Nucleotides
Staurosporine
Protein C Inhibitor
Phosphoric Diester Hydrolases
Protein Kinase Inhibitors
Adenylyl Cyclases
Albumins
Carrier Proteins
Down-Regulation
Lipids

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Serum-induced sensitization of cyclic AMP accumulation in C62B rat glioma cells. / Toews, Myron Lee; Hoffman, J. M.; Liewer, S. A.; Arneson-Rotert, L. J.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 262, No. 2, 01.01.1992, p. 471-478.

Research output: Contribution to journalArticle

Toews, Myron Lee ; Hoffman, J. M. ; Liewer, S. A. ; Arneson-Rotert, L. J. / Serum-induced sensitization of cyclic AMP accumulation in C62B rat glioma cells. In: Journal of Pharmacology and Experimental Therapeutics. 1992 ; Vol. 262, No. 2. pp. 471-478.
@article{355604e697e644b5847bc436f86930e4,
title = "Serum-induced sensitization of cyclic AMP accumulation in C62B rat glioma cells",
abstract = "Exposure of C62B rat glioma cells to fresh medium containing fetal bovine serum induced a sensitization of the subsequent ability of isoproterenol and forskolin to stimulate cyclic AMP accumulation, compared to cells exposed to fresh medium without serum. Isoproterenol stimulation was typically increased by 2- to 4-fold and forskolin stimulation by 3- to 5-fold. Sensitization occurred rapidly, was rapidly reversible and appeared to result from an increase in maximal stimulation. A commercial preparation of albumin, purified chromatographically so as to retain bound lipids and other factors, was able to mimic the effect of serum. In contrast to the effects of serum, exposure of cells to phorbol 12-myristate, 13-acetate induced little or no change in forskolin stimulation but a marked desensitization of isoproterenol stimulation that was due primarily to a decrease in potency. Neither the protein kinase C inhibitor staurosporine or overnight exposure to phorbol 12- myristate, 13-acetate to down-regulate protein kinase C prevented serum- induced sensitization. Pertussis toxin almost completely blocked serum- induced sensitization, suggesting involvement of a pertussis toxin-sensitive guanine nucleotide-binding protein in mediating the effects of serum. Sensitization was poorly retained in membrane adenylate cyclase assays. Studies with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, direct assays of cyclic AMP degradation by intact cells and assays of phosphodiesterase activity in cell lysates all indicated that degradation of cyclic AMP was decreased in serum-pretreated cells. Thus, both increased cyclic AMP synthesis and decreased cyclic AMP degradation may contribute to sensitization in these cells.",
author = "Toews, {Myron Lee} and Hoffman, {J. M.} and Liewer, {S. A.} and Arneson-Rotert, {L. J.}",
year = "1992",
month = "1",
day = "1",
language = "English (US)",
volume = "262",
pages = "471--478",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

TY - JOUR

T1 - Serum-induced sensitization of cyclic AMP accumulation in C62B rat glioma cells

AU - Toews, Myron Lee

AU - Hoffman, J. M.

AU - Liewer, S. A.

AU - Arneson-Rotert, L. J.

PY - 1992/1/1

Y1 - 1992/1/1

N2 - Exposure of C62B rat glioma cells to fresh medium containing fetal bovine serum induced a sensitization of the subsequent ability of isoproterenol and forskolin to stimulate cyclic AMP accumulation, compared to cells exposed to fresh medium without serum. Isoproterenol stimulation was typically increased by 2- to 4-fold and forskolin stimulation by 3- to 5-fold. Sensitization occurred rapidly, was rapidly reversible and appeared to result from an increase in maximal stimulation. A commercial preparation of albumin, purified chromatographically so as to retain bound lipids and other factors, was able to mimic the effect of serum. In contrast to the effects of serum, exposure of cells to phorbol 12-myristate, 13-acetate induced little or no change in forskolin stimulation but a marked desensitization of isoproterenol stimulation that was due primarily to a decrease in potency. Neither the protein kinase C inhibitor staurosporine or overnight exposure to phorbol 12- myristate, 13-acetate to down-regulate protein kinase C prevented serum- induced sensitization. Pertussis toxin almost completely blocked serum- induced sensitization, suggesting involvement of a pertussis toxin-sensitive guanine nucleotide-binding protein in mediating the effects of serum. Sensitization was poorly retained in membrane adenylate cyclase assays. Studies with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, direct assays of cyclic AMP degradation by intact cells and assays of phosphodiesterase activity in cell lysates all indicated that degradation of cyclic AMP was decreased in serum-pretreated cells. Thus, both increased cyclic AMP synthesis and decreased cyclic AMP degradation may contribute to sensitization in these cells.

AB - Exposure of C62B rat glioma cells to fresh medium containing fetal bovine serum induced a sensitization of the subsequent ability of isoproterenol and forskolin to stimulate cyclic AMP accumulation, compared to cells exposed to fresh medium without serum. Isoproterenol stimulation was typically increased by 2- to 4-fold and forskolin stimulation by 3- to 5-fold. Sensitization occurred rapidly, was rapidly reversible and appeared to result from an increase in maximal stimulation. A commercial preparation of albumin, purified chromatographically so as to retain bound lipids and other factors, was able to mimic the effect of serum. In contrast to the effects of serum, exposure of cells to phorbol 12-myristate, 13-acetate induced little or no change in forskolin stimulation but a marked desensitization of isoproterenol stimulation that was due primarily to a decrease in potency. Neither the protein kinase C inhibitor staurosporine or overnight exposure to phorbol 12- myristate, 13-acetate to down-regulate protein kinase C prevented serum- induced sensitization. Pertussis toxin almost completely blocked serum- induced sensitization, suggesting involvement of a pertussis toxin-sensitive guanine nucleotide-binding protein in mediating the effects of serum. Sensitization was poorly retained in membrane adenylate cyclase assays. Studies with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, direct assays of cyclic AMP degradation by intact cells and assays of phosphodiesterase activity in cell lysates all indicated that degradation of cyclic AMP was decreased in serum-pretreated cells. Thus, both increased cyclic AMP synthesis and decreased cyclic AMP degradation may contribute to sensitization in these cells.

UR - http://www.scopus.com/inward/record.url?scp=0026756916&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026756916&partnerID=8YFLogxK

M3 - Article

C2 - 1380077

AN - SCOPUS:0026756916

VL - 262

SP - 471

EP - 478

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 2

ER -