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Cell surface measurements of ATP release from single pancreatic β cells using a novel biosensor technique
Authors:Akihiro Hazama  Seiji Hayashi  Y. Okada
Affiliation:(1) Department of Cellular and Molecular Physiology, National Institute for Physiological Sciences, Okazaki 444–8585; CREST, Japan Science and Technology Corporation, Japan e-mail: okada@nips.ac.jp Tel.: +81-564-557731, Fax: +81-564-557735, JP
Abstract: To examine the possibility that ATP modulates insulin secretion by an autocrine mechanism, we measured the local concentration of released ATP at the surface of a single pancreatic β cell by a new biosensor technique, using PC12 cells expressing ligand-gated cation channels, P2X2 receptors. Upon application of glucose or glibenclamide, a series of current spikes, whose amplitude equates to an ATP concentration of over 25 μM, were recorded from a PC12 cell using the whole-cell patch-clamp technique, when placed near a rat pancreatic β cell at 37°C. The current response was inhibited by cooling (below 30°C) or by applying an ATP-hydrolysing enzyme (apyrase) or a P2 receptor blocker (suramin). Thus, it is concluded that pancreatic β cells secrete ATP in response to glucose stimulation, thereby increasing the ATP concentration close to the cell surface sufficiently high enough to enhance insulin secretion from the pancreatic β cells. Received: 8 May 1998 / Received after revision: 4 August 1998 / Accepted: 10 August 1998
Keywords:  ATP release  Autocrine  Biosensor  Exocytosis  Insulin secretion  P2 receptor  Pancreatic β   cell  PC12 cell
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