Glucose sensing mechanism has been intensively studied in pancreatic cells and neurons. Depolarization of membrane potential by closure of K-ATP. Kv and TASK channel, and subsequently Ca2+ entry via L-type voltage gated Ca2+ channel (VGCC) are implicated to mediate the signal transduction in these cells. However, the mechanism of non-excitable cells, which are lacking VGCC, for sensing glucose remains unclear. In this study, we utilized the calcium ratio measurement and patch clamping technique to study the effects of low glucose on [Ca2+](i) and currents in the human embryonic kidney epithelial cells (HEK 293). We found low glucose evoked a significant reversible [Ca2+](i) elevation in HEK 293 independent of the closure of Kv channels. This increase of [Ca2+](i) was mediated by Ca2+ entry across plasma membrane and exhibited a dosage dependent behaviour to external glucose concentration. The low glucose-induced entry of Ca2+ was characterized as a voltage independent behaviour and had cation permeability to Na+ and Ca2+. The modulation of PLC. AMPK. tyrosine kinase and cADPribose failed to regulate this glucose-sensitive Ca2+ entry. In addition, the entry of Ca2+ was insensitive to nifedipine, 2APB, SKF, La3+, Gd3+, and KBR9743, suggesting a novel signal pathway in mediating glucose sensing. (c) 2007 Elsevier Inc. All rights reserved.