Modulation of hepatocellular swelling-activated K+currents by phosphoinositide pathway-dependent protein kinase C

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Abstract
K+channels participate in the regulatory volume decrease (RVD) accompanying hepatocellular nutrient uptake and bile formation. We recently identified KCNQ1 as a molecular candidate for a significant fraction of the hepatocellular swelling-activated K+current ( IKVol). We have shown that the KCNQ1 inhibitor chromanol 293B significantly inhibited RVD-associated K+flux in isolated perfused rat liver and used patch-clamp techniques to define the signaling pathway linking swelling to IKVolactivation. Patch-electrode dialysis of hepatocytes with solutions that maintain or increase phosphatidylinositol 4,5-bisphosphate (PIP2) increased IKVol, whereas conditions that decrease cellular PIP2decreased IKVol. GTP and AlF4−stimulated IKVoldevelopment, suggesting a role for G proteins and phospholipase C (PLC). Supporting this, the PLC blocker U-73122 decreased IKVoland inhibited the stimulatory response to PIP2or GTP. Protein kinase C (PKC) is involved, because K+current was enhanced by 1-oleoyl-2-acetyl- sn-glycerol and inhibited after chronic PKC stimulation with phorbol 12-myristate 13-acetate (PMA) or the PKC inhibitor GF 109203X. Both IKVoland the accompanying membrane capacitance increase were blocked by cytochalasin D or GF 109203X. Acute PMA did not eliminate the cytochalasin D inhibition, suggesting that PKC-mediated IKVolactivation involves the cytoskeleton. Under isotonic conditions, a slowly developing K+current similar to IKVolwas activated by PIP2, lipid phosphatase inhibitors to counter PIP2depletion, a PLC-coupled α1-adrenoceptor agonist, or PKC activators and was depressed by PKC inhibition, suggesting that hypotonicity is one of a set of stimuli that can activate IKVolthrough a PIP2/PKC-dependent pathway. The results indicate that PIP2indirectly activates hepatocellular KCNQ1-like channels via cytoskeletal rearrangement involving PKC activation.

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