Continuous monitoring of ATP levels in living insulin secreting cells expressing cytosolic firefly luciferase

FEBS Letters

Wollheim, Claes B¹  Wang, Haiyan¹  Maechler, Pierre¹ 

[1] Division de Biochimie Clinique, Department of Internal Medicine, University Medical Centre, CH-1211 Geneva 4, Switzerland

关键词: ATP;    Luciferase;    Insulin secretion;    INS-1 cell;    Tetracycline transactivator;    ADP-ATP translocase;    met-Suc;    methyl-succinate;    KRBH;    Krebs-Ringer bicarbonate HEPES buffer;    FCCP;    carbonyl cyanide p-trifluoromethoxyphenylhydrazone;   

DOI  :  10.1016/S0014-5793(97)01618-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The second messenger role of ATP in insulin secretion was investigated in living INS-1 insulinoma cells. ATP-dependent luminescence was monitored in cells expressing high levels of firefly luciferase under the control of the tetracycline-dependent transactivator. The calibration of luminescence in permeabilized cells yielded similar ATP levels as those obtained in cell extracts with a conventional ATP assay. Stimulation of insulin secretion by glucose or methyl-succinate was correlated with rises of cellular ATP in simultaneous measurements. ATP generation was decreased by inhibition of the ADP-ATP translocase. This approach demonstrates the feasibility of defining the dynamic relationship between ATP and other parameters involved in metabolism-secretion coupling.

【 授权许可】

Unknown