Kv7.1 surface expression is regulated by epithelial cell polarization

Abstract

The potassium channel KV7.1 is expressed in the heart where it contributes to the repolarization of the cardiac action potential. In addition, KV7.1 is expressed in epithelial tissues where it plays a role in salt and water transport. Mutations in the kcnq1 gene can lead to long QT syndrome and deafness, and several mutations have been described as trafficking mutations. To learn more about the basic mechanisms that regulate KV7.1 surface expression, we have investigated the trafficking of KV7.1 during the polarization process of the epithelial cell line Madin-Darby Canine Kidney (MDCK) using a modified version of the classical calcium switch. We discovered that KV7.1 exhibits a very dynamic localization pattern during the calcium switch. When MDCK cells are kept in low calcium medium, KV7.1 is mainly observed at the plasma membrane. During the first hours of the switch, KV7.1 is removed from the plasma membrane and an intracellular accumulation in the endoplasmic reticulum (ER) is observed. The channel is retained in the ER until the establishment of the lateral membranes at which point KV7.1 is released from the ER and moves to the plasma membrane. Our data furthermore suggest that while the removal of KV7.1 from the cell surface and its accumulation in the ER could involve activation of protein kinase C, the subsequent release of KV7.1 from the ER depends on phosphoinositide 3-kinase (PI3K) activation. In conclusion, our results demonstrate that KV7.1 surface expression is regulated by signaling mechanisms involved in epithelial cell polarization in particular signaling cascades involving protein kinase C and PI3K.