Effect of ethanol on protein kinase Cζ and p70S6 kinase activation by carbachol: A possible mechanism for ethanol-induced inhibition of glial cell proliferation

Marina Guizzetti, Lucio G. Costa

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30 Citations (Scopus)

Abstract

The signal transduction pathways that mediate the mitogenic response of muscarinic acetylcholine receptors in astroglial cells have not been fully elucidated. In this study we investigated the activation of p70S6 kinase (p70S6K) by carbachol in 1321 N1 astroctyoma cells. Carbachol induced a dose- and time-dependent activation of p70S6K, as evidenced by increased phosphorylation at Thr-389, Thr-421 and Ser-424, by increased p70S6K activity, and by a shift in its molecular weight. Activation of p70S6K was mediated by M3 muscarinic acetylcholine receptors (mAChRs) and was inhibited by two phosphatidylinositol-3-kinase (P13-K) inhibitors, by a pseudosubstrate to protein kinase C (PKC) ζ, and by the p70S6K inhibitor rapamycin. Carbachol-induced DNA synthesis was strongly inhibited by rapamycin, suggesting that p70S6K activation plays an important role in carbachol-induced cell proliferation. Ethanol (25-100 mM) has been shown to inhibit carbachol-induced proliferation of astroglial cells. In the same range of concentrations, ethanol also inhibits carbacholinduced activation of PKCζ and of p70S6K. On the other hand, inhibition of P13-kinase was only observed at higher ethanol concentrations. These results indicate that activation of the PKCζ → p70S6K pathway by M3 mAChRs may play a role in the increased DNA synthesis and may represent a target for ethanol-induced inhibition of astroglial cell proliferation.

Original languageEnglish (US)
Pages (from-to)38-46
Number of pages9
JournalJournal of Neurochemistry
Volume82
Issue number1
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Cell proliferation
Carbachol
Neuroglia
Ethanol
Phosphotransferases
Chemical activation
Cell Proliferation
Muscarinic Receptors
Sirolimus
Phosphatidylinositol 3-Kinase
Signal transduction
Phosphorylation
DNA
protein kinase C kinase
Signal Transduction
Molecular Weight
Molecular weight

Keywords

  • Astrocytoma cells
  • Cell proliferation
  • Ethanol
  • Muscarinic receptors
  • P70S6K
  • PKCζ

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

@article{04822b8b67a44c30b14c6741428f6089,
title = "Effect of ethanol on protein kinase Cζ and p70S6 kinase activation by carbachol: A possible mechanism for ethanol-induced inhibition of glial cell proliferation",
abstract = "The signal transduction pathways that mediate the mitogenic response of muscarinic acetylcholine receptors in astroglial cells have not been fully elucidated. In this study we investigated the activation of p70S6 kinase (p70S6K) by carbachol in 1321 N1 astroctyoma cells. Carbachol induced a dose- and time-dependent activation of p70S6K, as evidenced by increased phosphorylation at Thr-389, Thr-421 and Ser-424, by increased p70S6K activity, and by a shift in its molecular weight. Activation of p70S6K was mediated by M3 muscarinic acetylcholine receptors (mAChRs) and was inhibited by two phosphatidylinositol-3-kinase (P13-K) inhibitors, by a pseudosubstrate to protein kinase C (PKC) ζ, and by the p70S6K inhibitor rapamycin. Carbachol-induced DNA synthesis was strongly inhibited by rapamycin, suggesting that p70S6K activation plays an important role in carbachol-induced cell proliferation. Ethanol (25-100 mM) has been shown to inhibit carbachol-induced proliferation of astroglial cells. In the same range of concentrations, ethanol also inhibits carbacholinduced activation of PKCζ and of p70S6K. On the other hand, inhibition of P13-kinase was only observed at higher ethanol concentrations. These results indicate that activation of the PKCζ → p70S6K pathway by M3 mAChRs may play a role in the increased DNA synthesis and may represent a target for ethanol-induced inhibition of astroglial cell proliferation.",
keywords = "Astrocytoma cells, Cell proliferation, Ethanol, Muscarinic receptors, P70S6K, PKCζ",
author = "Marina Guizzetti and Costa, {Lucio G.}",
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T1 - Effect of ethanol on protein kinase Cζ and p70S6 kinase activation by carbachol

T2 - A possible mechanism for ethanol-induced inhibition of glial cell proliferation

AU - Guizzetti, Marina

AU - Costa, Lucio G.

PY - 2002

Y1 - 2002

N2 - The signal transduction pathways that mediate the mitogenic response of muscarinic acetylcholine receptors in astroglial cells have not been fully elucidated. In this study we investigated the activation of p70S6 kinase (p70S6K) by carbachol in 1321 N1 astroctyoma cells. Carbachol induced a dose- and time-dependent activation of p70S6K, as evidenced by increased phosphorylation at Thr-389, Thr-421 and Ser-424, by increased p70S6K activity, and by a shift in its molecular weight. Activation of p70S6K was mediated by M3 muscarinic acetylcholine receptors (mAChRs) and was inhibited by two phosphatidylinositol-3-kinase (P13-K) inhibitors, by a pseudosubstrate to protein kinase C (PKC) ζ, and by the p70S6K inhibitor rapamycin. Carbachol-induced DNA synthesis was strongly inhibited by rapamycin, suggesting that p70S6K activation plays an important role in carbachol-induced cell proliferation. Ethanol (25-100 mM) has been shown to inhibit carbachol-induced proliferation of astroglial cells. In the same range of concentrations, ethanol also inhibits carbacholinduced activation of PKCζ and of p70S6K. On the other hand, inhibition of P13-kinase was only observed at higher ethanol concentrations. These results indicate that activation of the PKCζ → p70S6K pathway by M3 mAChRs may play a role in the increased DNA synthesis and may represent a target for ethanol-induced inhibition of astroglial cell proliferation.

AB - The signal transduction pathways that mediate the mitogenic response of muscarinic acetylcholine receptors in astroglial cells have not been fully elucidated. In this study we investigated the activation of p70S6 kinase (p70S6K) by carbachol in 1321 N1 astroctyoma cells. Carbachol induced a dose- and time-dependent activation of p70S6K, as evidenced by increased phosphorylation at Thr-389, Thr-421 and Ser-424, by increased p70S6K activity, and by a shift in its molecular weight. Activation of p70S6K was mediated by M3 muscarinic acetylcholine receptors (mAChRs) and was inhibited by two phosphatidylinositol-3-kinase (P13-K) inhibitors, by a pseudosubstrate to protein kinase C (PKC) ζ, and by the p70S6K inhibitor rapamycin. Carbachol-induced DNA synthesis was strongly inhibited by rapamycin, suggesting that p70S6K activation plays an important role in carbachol-induced cell proliferation. Ethanol (25-100 mM) has been shown to inhibit carbachol-induced proliferation of astroglial cells. In the same range of concentrations, ethanol also inhibits carbacholinduced activation of PKCζ and of p70S6K. On the other hand, inhibition of P13-kinase was only observed at higher ethanol concentrations. These results indicate that activation of the PKCζ → p70S6K pathway by M3 mAChRs may play a role in the increased DNA synthesis and may represent a target for ethanol-induced inhibition of astroglial cell proliferation.

KW - Astrocytoma cells

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