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

FEBS Lett. 1998 Feb 6;422(3):328-32. doi: 10.1016/s0014-5793(97)01618-9.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Animals
  • Biological Transport
  • Coleoptera
  • Cytosol / metabolism
  • Insulin / metabolism*
  • Insulin Secretion
  • Islets of Langerhans / metabolism*
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Tumor Cells, Cultured


  • Insulin
  • Recombinant Proteins
  • Adenosine Triphosphate
  • Luciferases