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J Biol Chem, Vol. 274, Issue 43, 30979-30986, October 22, 1999
From the ATP stimulates Cl
The Lipid Products of Phosphoinositide 3-Kinase Contribute to
Regulation of Cholangiocyte ATP and Chloride Transport
,,,,
Departments of Pediatrics and Medicine,
Children's Hospital and the University of Colorado Health Sciences
Center, Denver, Colorado 80262 and the ¶ Signal Transduction
Group, Boston Biomedical Research Group,
Boston, Massachusetts 02114
secretion
and bile formation by activation of purinergic receptors in the apical
membrane of cholangiocytes. The purpose of these studies was to
determine the cellular origin of biliary ATP and to assess the
regulatory pathways involved in its release. In Mz-Cha-1 human
cholangiocarcinoma cells, increases in cell volume were followed by
increases in phophoinositide (PI) 3-kinase activity, ATP release, and
membrane Cl permeability. PI 3-kinase signaling appears
to play a regulatory role because ATP release was inhibited by
wortmannin or LY294002 and because volume-sensitive current activation
was inhibited by intracellular dialysis with antibodies to the 110 kDa-subunit of PI 3-kinase. Similarly, in intact normal rat
cholangiocyte monolayers, increases in cell volume stimulated luminal
Cl secretion through a wortmannin-sensitive pathway. To
assess the role of PI 3-kinase more directly, cells were dialyzed with
the synthetic lipid products of PI 3-kinase. Intracellular delivery of
phosphatidylinositol 3,4-bisphosphate, and phosphatidylinositol 3,4,5-trisphosphate activated Cl currents analogous to
those observed following cell swelling. Taken together, these findings
indicate that volume-sensitive activation of PI 3-kinase and the
generation of lipid messengers modulate cholangiocyte ATP release,
Cl secretion, and, hence, bile formation.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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