Orexin neurons are specifically localized in and around the lateral hypothalamus (LH), a feeding center. Intracerebroventricular administration of orexin-A and -B stimulates feeding as well as arousal. However, little is known regarding the regulators of the orexin neuron activity. The neurons that are activated under low glucose conditions, glucose-sensitive neurons, are located in the LH and have been implicated in the control of feeding. The present study investigated the effect of glucose on the single orexin neurons isolated from the rat LH, by measuring cytosolic Ca(2+) concentration ([Ca(2+)](i)) by fura-2 microfluorometry followed by immunocytochemical staining with anti-orexin antiserum. A shift of glucose concentration form 8.3 to 2.8 mM in the superfusion solution increased [Ca(2+)](i) in 13 out of 32 orexin-immunoreactive LH neurons. The results demonstrate that glucose-sensitive orexin neurons are present in the LH and that these neurons may play a role in linking the metabolic state in the body to the orexigenic, and could also, awakening signaling in the brain.