Bacterial endotoxin alters kinetics of BK channels in rat cerebrovascular smooth muscle cells

Abstract

Patch-clamp recordings were used to study the effects of Escherichia coli bacterial endotoxin (lipopolysaccharide, LPS) on the properties of large-conductance, Ca 2+-dependent K + channels (BK channels) in the membrane of enzymatically dispersed rat cerebrovascular smooth muscle cells (CVSMCs). LPS had negligible effects on the kinetic and conductance properties of BK channels when applied to the extracellular domain of these channels. However, acute application of LPS (10–100 μg/ml) to inside-out patches of CVSMC membrane isolated in a cell-free environment rapidly and reversibly increased the open probability of BK channels, leaving the conductance of these channels unaltered. The magnitude of this effect decreased as the concentration of free Ca 2+ at the cytoplasmic membrane face was lowered, but was little affected by changes in membrane potential. Kinetic analysis showed that LPS accelerated reopening of BK channels while having little effect on mean channel open time. Detoxified E. coli LPS, from which the fatty acid chains of Lipid A were partially removed, showed slightly reduced activity when compared to the parent endotoxin molecule. A purified E. coli Lipid A had negligible effects on BK channel function. These results indicate that LPS activates BK channels in cerebrovascular smooth muscle cells when present at the cytoplasmic membrane face. This novel mechanism may provide insights into the regulation of BK channels by intracellular, membrane-associated elements.