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(Received for publication, April 25, 1997)
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From the In a model liver cell line, recovery from
swelling is mediated by a sensitive autocrine pathway involving
conductive release of ATP, P2 receptor stimulation,
and opening of membrane Cl Division of Gastroenterology, Department of
Medicine, University of Colorado Health Sciences Center, Denver,
Colorado 80262, the Division of Gastroenterology, Department of
Medicine, University of California, San Francisco, California
94143-0538, and the ¶ Division of Gastroenterology, Department of
Medicine, Duke University Medical Center,
Durham, North Carolina 27710
channels (Wang, Y., Roman,
R. M., Lidofsky, S. D., and Fitz, J. G. (1996)
Proc. Natl. Acad. Sci. U. S. A. 93, 12020-12025). However, the mechanisms coupling changes in cell volume to ATP release
are not known. Based on evidence that certain ATP-binding cassette
(ABC) proteins may function as ATP channels or channel regulators, we
evaluated the potential role of ABC proteins by comparing ATP release
and volume regulation in rat HTC and HTC-R hepatoma cells, the latter
of which overexpress Mdr proteins. In both cell types, Cl
current activation (ICl-swell) and volume
recovery following swelling were dependent on conductive ATP efflux.
The rate of volume recovery was ~6-fold faster in HTC-R cells
compared with HTC cells. This effect is likely due to enhanced ABC
protein-dependent ATP release since (i)
ICl-swell and cell volume recovery were eliminated by inhibition of P-glycoprotein transport (20 µM verapamil and 15 µM cyclosporin A); (ii)
swelling-induced Cl current density was similar in
both cell types (approximately 
50 pA/pF; not significant); and (iii)
ATP conductance measured by whole-cell techniques was increased
~3-fold in HTC-R cells compared with HTC cells. Moreover, HTC-R cells
exhibited enhanced survival during hypotonic stress. By modulating ATP
release, hepatic ABC proteins may play a key role in the cellular
pathways coupling changes in cell volume to ion permeability and
secretion.
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