ATP-sensitive K channel modulation by products of PLA2 action in the insulin-secreting HIT cell line

Abstract

Insulin secretion from the islets of Langerhans may be initiated or potentiated by increased phospholipase A2 (PLA2) activity. This patch-clamp study of the insulin-secreting HIT tumor cell line assessed whether inhibition of the ATP-sensitive potassium (KATP) channel, which modulates the secretion-associated beta-cell electrical activity, contributes to the secretory response to PLA2. Exogenous PLA2 (100-1,000 mU/ml) reversibly suppressed KATP channel activity in excised inside-out patches. Similarly, mellitin (0.5-5 micrograms/ml), a bee venom component that increases phospholipid susceptibility to metabolism by PLA2, suppressed KATP channel activity, suggesting that PLA2 is present in excised patches. Adding low concentrations of particular lysophospholipids or arachidonic acid also reduced KATP channel activity in excised inside-out patches. In cell-attached patches, the lysophospholipids had a similar effect, whereas arachidonic acid caused channel stimulation; this latter effect was reversed by cyclooxygenase inhibitors. A recently identified ATP-stimulated PLA2 in beta-cells has been proposed as an important mediator of stimulus-secretion coupling in response to nutrients. The present data illustrating that initial products of PLA2 action on membrane phospholipids reduce KATP channel activity indicate a mechanism that may contribute an early stimulatory signal in this pathway. The observation that metabolism of arachidonate via the cyclooxygenase pathway causes KATP channel stimulation demonstrates a potential counterregulatory mechanism.