Characterization of calcium oscillations in normal and benzo[a]pyrene-treated clone 9 cells

Toxicol Sci. 2002 Aug;68(2):444-50. doi: 10.1093/toxsci/68.2.444.

Abstract

Intracellular Ca2+ oscillations induced by oxytocin and vasopressin were analyzed in a rat liver cell line (Clone 9) in order to identify mechanisms by which benzo[a]pyrene (BaP) alters Ca2+ signaling patterns in these cells. Clone 9 cells exhibit an initial Ca2+ spike, followed by Ca2+ oscillations upon oxytocin or vasopressin treatment. The range of frequencies (maximum 110 mHz) was dependent on agonist concentration with a constant amplitude less than or equal to the amount of Ca2+ generated from the inositol trisphosphate (InsP(3))-sensitive pool. This study examined contributions of extracellular and intracellular pools to the frequency of Ca2+ oscillations and the role of membrane channels, second messengers, and different pharmacological reagents on the regulation of oscillation frequency in both control and BaP-treated cells. Results indicated that the Ca2+ oscillations are mainly due to inositol 1,4,5-triphosphate (InsP(3))-sensitive stores and that extracellular Ca2+ contributes to refilling of this intracellular Ca2+ pool. The frequency of Ca2+ oscillations is also sharply affected by protein kinase C activated by phospholipase C. In BaP-treated Clone 9 cells, basal Ca2+ levels were elevated and the frequency of Ca2+ oscillations was suppressed in a dose-dependent fashion. Suppression of Ca2+ oscillations is due, at least in part, to an effect of BaP on enhanced opening of K+ channels. This was confirmed by showing that inhibition of the K+ channel opening by tetraethylammonium chloride can reverse the effect of BaP on oxytocin-induced Ca2+ oscillations, and potentially decrease the toxicity of BaP.

Publication types

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

MeSH terms

  • Animals
  • Benzo(a)pyrene / toxicity*
  • Calcium / metabolism*
  • Calcium Signaling*
  • Carcinogens / toxicity*
  • Cell Line
  • Clone Cells
  • Drug Antagonism
  • Image Cytometry
  • Liver / cytology
  • Liver / drug effects*
  • Liver / metabolism*
  • Microscopy, Fluorescence
  • Oxytocin / pharmacology
  • Potassium Channel Blockers / pharmacology
  • Rats
  • Tetraethylammonium / pharmacology
  • Vasopressins / pharmacology

Substances

  • Carcinogens
  • Potassium Channel Blockers
  • Vasopressins
  • Benzo(a)pyrene
  • Oxytocin
  • Tetraethylammonium
  • Calcium