ATP-sensitive potassium (K ATP ) channels play a central role in glucose-stimulated insulin secretion (GSIS) by pancreatic β-cells. Activity of these channels is determined by their open probability (P o ) and the number of channels present in a cell. Glucose is known to reduce P o , but whether it also affects the channel density is unknown. Using INS-1 model β-cell line, we show that the expression of K ATP channel subunits, Kir6.2 and SUR1, is high at low glucose, but declines sharply when the ambient glucose concentration exceeds 5mM. In response to glucose deprivation, channel synthesis increases rapidly by up-regulating translation of existing mRNAs. The effects of glucose deprivation could be mimicked by pharmacological activation of 5′-AMP-activated protein kinase with 5-aminoimidazole-4-carboxamide ribonucleotide and metformin. Pancreatic β-cells which have lost their ability for GSIS do not show such changes implicating a possible (patho-)physiological link between glucose-regulated K ATP channel expression and the capacity for normal GSIS.