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

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Abstract
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.