Real-time measurements of acetylcholine-induced release of ATP from bovine medullary chromaffin cells

FEBS Lett. 1985 Jun 17;185(2):323-7. doi: 10.1016/0014-5793(85)80931-5.

Abstract

The luminescent oxidation of luciferin has been used to monitor acetylcholine-induced ATP release from cultured bovine chromaffin cells. Acetylcholine (1-100 microM) evoked ATP release of up to 30% of the total cellular ATP. This secretion required external free calcium and could also be elicited by K+-induced membrane depolarization. The size of the cytosolic ATP compartment was estimated as 5% of the ATP in the cell by solubilising the cell membrane using digitonin (20 microM) or by application to the cells of brief pulses (2 microseconds) of high electric field (2000 V/cm). Blockers of the voltage-gated Ca2+ channel effectively blocked K+-induced ATP release, while the acetylcholine antagonists d-tubocurarine and beta-bungarotoxin inhibited the acetylcholine-induced release of ATP. These data support the concept that ATP is released together with the catecholamines by exocytosis of chromaffin granule contents.

MeSH terms

  • Acetylcholine / pharmacology*
  • Adenosine Triphosphate / metabolism*
  • Adrenal Medulla / metabolism*
  • Animals
  • Calcium / physiology
  • Cattle
  • Cell Membrane Permeability
  • Chromaffin Granules / metabolism*
  • Chromaffin System / metabolism*
  • Cytosol / metabolism
  • Digitonin
  • Firefly Luciferin
  • Luciferases
  • Receptors, Nicotinic / drug effects

Substances

  • Receptors, Nicotinic
  • Firefly Luciferin
  • Adenosine Triphosphate
  • Luciferases
  • Digitonin
  • Acetylcholine
  • Calcium