Insulin activates ATP-sensitive K+ channels in pancreatic beta-cells through a phosphatidylinositol 3-kinase-dependent pathway

Insulin is known to regulate pancreatic beta -cell function through the act ivation of cell surface insulin receptors, phosphorylation of insulin recep tor substrate (IRS)-1 and -2, and activation of phosphatidylinositol (PI) 3 -kinase. However, an acute effect of insulin in modulating beta -cell elect rical activity and its underlying ionic currents has not been reported. Usi ng the perforated patch clamp technique, we found that insulin (1-600 nmol/ l) but not IGF-1 (100 nmol/l) reversibly hyperpolarized single mouse beta - cells and inhibited their electrical activity. The dose-response relationsh ip for insulin yielded a maximal change (mean +/- SE) in membrane potential of -13.6 +/- 2.0 mV (P < 0.001) and a 50% effective dose of 25.9 +/- 0.1 n mol/l (n = 63). Exposing patched beta -cells within intact islets to 200 nm ol/l insulin produced similar results, hyperpolarizing islets from -47.7 +/ - 3.3 to -65.6 +/- 3.7 mV (P < 0.0001, n = 11). In single cells, insulin-in duced hyperpolarization was associated with a threefold increase in whole-c ell conductance from 0.6 +/- 0.1 to 1.7 +/- 0.2 nS (P < 0.001, n = 10) and a shift in the current reversal potential from -25.7 +/- 2.5 to -63.7 +/- 1 .0 mV (P < 0.001 vs. control, n = 9; calculated K+ equilibrium potential = -90 mV). The effects of insulin were reversed by tolbutamide, which decreas ed cell conductance to 0.5 0.1 nS and shifted the current reversal potentia l to -25.2 +/- 2.3 mV. Insulin-induced P-cell hyperpolarization was suffici ent to abolish intracellular calcium concentration ([Ca2+](i)) oscillations measured in pancreatic islets exposed to 10 mmol/l glucose. The applicatio n of 100 nmol/l wortmannin to inactivate PI 3-kinase, a key enzyme in insul in signaling, was found to reverse the effects of 100 nmol/l insulin. In ce ll-attached patches, single AIT-sensitive K+ (K-ATP) channels were activate d by bath-applied insulin and subsequently inhibited by wortmannin. Our dat a thus demonstrate that insulin activates the K-ATP channels of single mous e pancreatic beta -cells and islets, resulting in membrane hyperpolarizatio n, an inhibition of electrical activity, and the abolition of [Ca2+](i) osc illations . We thus propose that locally released insulin might serve as a negative feedback signal within the islet under physiological conditions.