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1 Department of Medicine, University of Colorado Health Sciences Center, Denver, Colorado 80262; and 2 Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, Alabama 35294
Extracellular
ATP functions as an important autocrine and paracrine signal that
modulates a broad range of cell and organ functions through activation
of purinergic receptors in the plasma membrane. Because little is known
of the cellular mechanisms involved in ATP release, the purpose of
these studies was to evaluate the potential role of the lanthanide
Gd3+ as an inhibitor of ATP
permeability and to assess the physiological implications of impaired
purinergic signaling in liver cells. In rat hepatocytes and HTC
hepatoma cells, increases in cell volume stimulate ATP release, and the
localized increase in extracellular ATP increases membrane
Cl
permeability and
stimulates cell volume recovery through activation of
P2 receptors. In cells in culture,
spontaneous ATP release, as measured by a luciferin-luciferase-based
assay, was always detectable under control conditions, and
extracellular ATP concentrations increased 2- to 14-fold after
increases in cell volume. Gd3+
(200 µM) inhibited volume-sensitive
ATP release by >90% (P < 0.001),
inhibited cell volume recovery from swelling
(P < 0.01), and uncoupled cell
volume from increases in membrane
Cl permeability
(P < 0.01). Moreover,
Gd3+ had similar inhibitory
effects on ATP release from other liver and epithelial cell models.
Together, these findings support an important physiological role for
constitutive release of ATP as a signal coordinating cell volume and
membrane ion permeability and suggest that
Gd3+ might prove to be an
effective inhibitor of ATP-permeable channels once they are identified.
cell volume; liver; chloride channel
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