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FEBS Lett. 1998 Feb 6;422(3):328-32.

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

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1
Department of Internal Medicine, University Medical Centre, Geneva, Switzerland.

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.

PMID:
9498809
DOI:
10.1016/s0014-5793(97)01618-9
[Indexed for MEDLINE]
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