Regulation of cation channels in liver cells by intracellular calcium and protein kinase C

J. G. Fitz, A. H. Sostman, J. P. Middleton

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

The regulation of Ca2+-permeant cation channels in HTC hepatoma cells was investigated using patch clamp and fluorescence techniques. In intact cells, exposure to nucleotide analogues ATP, uridine 5'-triphosphate (UTP), and adenosine 5'-O-(3-thiotriphosphate) (ATPγS) caused transient opening of channels with linear conductances of ~18 and ~28 pS. Channels were permeable to Na+, K+, and Ca2+ and carried inward (depolarizing) current at the resting potential. Exposure to thapsigargin to increase cytosolic Ca2+ concentration ([Ca2+](i)) opened similar channels, suggesting that opening is stimulated by a rise in [Ca2+](i). In subconfluent monolayers, ATP increased [Ca2+](i) with half-maximal effects at ~7.4 μM; at 10-4 M, the peak increase in [Ca2+](i) was ATP > UTP > ATPγS > > 2-methyl- thioadenosine 5'-triphosphate, α,β-methyleneadenosine 5'-triphosphate, and adenosine. The relative potency suggests that the effects are mediated by 5'- nucleotide receptors. In excised inside-out patches, channels were not activated by myo-inositol 1,4,5-trisphosphate (50-100 μM) or myo-inositol 1,3,4,5-trisphosphate (20 μM) but opened after increases in Ca2+ to greater than ~250 nM, consistent with a direct role for Ca2+ in channel opening. In intact cells, channel opening was followed by a prolonged refractory period. Protein kinase C appears to contribute by inhibition of the ATP-stimulated [Ca2+](i) response and by direct inhibitory effects on the channel. These findings indicate that extracellular ATP leads to modulation of liver cell cation channels through activation of 5'-nucleotide receptors and are consistent with a model in which transient opening of channels is stimulated by a rise in [Ca2+](i) and subsequent closure is mediated by protein kinase C-dependent pathways.

Original languageEnglish (US)
Pages (from-to)G677-G684
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume266
Issue number4 29-4
DOIs
StatePublished - 1994

Keywords

  • ATP
  • hepatocyte
  • ion channel
  • purinergic

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

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