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Am J Physiol Gastrointest Liver Physiol 282: G116-G122, 2002;
0193-1857/02 $5.00
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Vol. 282, Issue 1, G116-G122, January 2002

Molecular characterization of volume-sensitive SKCa channels in human liver cell lines

Richard Roman1, Andrew P. Feranchak2, Marlyn Troetsch1, Jeffrey C. Dunkelberg1, Gordon Kilic1, Thorsten Schlenker3, Jerome Schaack4, and J. Gregory Fitz1

Departments of 1 Medicine, 2 Pediatrics, and 4 Microbiology, School of Medicine, University of Colorado Health Sciences Center, Denver, Colorado 80262; and 3 Department of Medicine, Ruprecht-Karls-Universität, D-69117 Heidelberg, Germany

In human liver, Ca2+-dependent changes in membrane K+ permeability play a central role in coordinating functional interactions between membrane transport, metabolism, and cell volume. On the basis of the observation that K+ conductance is partially sensitive to the bee venom toxin apamin, we aimed to assess whether small-conductance Ca2+-sensitive K+ (SKCa) channels are expressed endogenously and contribute to volume-sensitive K+ efflux and cell volume regulation. We isolated a full-length 2,140-bp cDNA (hSK2) highly homologous to rat brain rSK2 cDNA, including the putative apamin-sensitive pore domain, from a human liver cDNA library. Identical cDNAs were isolated from primary human hepatocytes, human HuH-7 hepatoma cells, and human Mz-ChA-1 cholangiocarcinoma cells. Transduction of Chinese hamster ovary cells with a recombinant adenovirus encoding the hSK2-green fluorescent protein fusion construct resulted in expression of functional apamin-sensitive K+ channels. In Mz-ChA-1 cells, hypotonic (15% less sodium glutamate) exposure increased K+ current density (1.9 ± 0.2 to 37.5 ± 7.1 pA/pF; P < 0.001). Apamin (10-100 nM) inhibited K+ current activation and cell volume recovery from swelling. Apamin-sensitive SKCa channels are functionally expressed in liver and biliary epithelia and likely contribute to volume-sensitive changes in membrane K+ permeability. Accordingly, the hSK2 protein is a potential target for pharmacological modulation of liver transport and metabolism through effects on membrane K+ permeability.

hepatocyte; cholangiocyte; cell volume; apamin


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