The Journal of Membrane Biology

, Volume 32, Issue 1, pp 201–230 | Cite as

Intracellular uptake and α-amylase and lactate dehydrogenase releasing actions of the divalent cation ionophore A23187 in dissociated pancreatic acinar cells

  • Douglas E. Chandler
  • John A. Williams


Intracellular uptake of A23187 and the increased release of amylase and lactate dehydrogenase (LDH) accompanying ionophore uptake was studied using dissociated acinar cells prepared from mouse pancreas. Easily detected changes in the fluorescence excitation spectrum of A23187 upon transfer of the ionophore from a Tris-buffered Ringer's to cell membranes were used to monitor A23187 uptake. Uptake was rapid in the absence of extracellular Ca2+ and Mg2+ (t1/2=1 min) and much slower in the presence of Ca2+ or Mg2+ (t1/2=20 min). Cell-associated ionophore was largely intracellular as indicated by fluorescence microscopy, lack of spectral sensitivity to changes in extracellular Ca2+ and Mg2+, and by equivalent interaction of ionophore with membranes of whole and sonicated cells.

A23187 (10 μm) increased amylase release 200% in the presence of extracellular Ca2+ and Mg2+. In the absence of Ca2+ (but in the presence of Mg2+) A23187 did not increase amylase release. A23187 (10 μm) also produced Ca2+-dependent cell damage, as judged by increased LDH release, increased permeability to trypan blue, and by disruption of cell morphology. The cell damaging and amylase releasing properties of A23187 were distinguished by their time course and dose-response relationship. A23187 (1 μm) increased amylase release 140% without increasing LDH release or permeability to trypan blue.


Amylase Lactate Dehydrogenase Trypan Blue Excitation Spectrum Acinar Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Amsterdam, A., Jamieson, J.D. 1972. Structural and functional characterization of isolated pancreatic exocrine cells.Proc. Nat. Acad. Sci. USA 69:3028PubMedGoogle Scholar
  2. Amsterdam, A., Jamieson, J.D. 1974. Studies on dispersed pancreatic exocrine cells. I. Dissociation technique and morphologic characteristics of separated cells.J. Cell Biol. 63:1037PubMedGoogle Scholar
  3. Ashby, J.P., Speake, R.N. 1975. Insulin and glucagon secretion from isolated islets of langerhans. Effects of calcium ionophores.Biochem. J. 150:89PubMedGoogle Scholar
  4. Binet, A., Volfin, P. 1975. Effect of the A23187 ionophore on mitochondrial Mg2+ and Ca2+.FEBS Lett. 49:400CrossRefPubMedGoogle Scholar
  5. Case, R.M., Clausen, T. 1973. The relationship between calcium exchange and enzyme secretion in the isolated rat pancreas.J. Physiol. (London) 235:75Google Scholar
  6. Case, G.D., Vanderkooi, J.M., Scarpa, A. 1974. Physical properties of biological membranes determined by the fluorescence of the calcium ionophore A23187.Arch. Biochem. Biophys. 162:174PubMedGoogle Scholar
  7. Caswell, A.H., Pressman, B.C. 1972. Kinetics of transport of divalent cations across sarcoplasmic reticulum vesicles induced by ionophores.Biochem. Biophys. Res. Commun. 49:292CrossRefPubMedGoogle Scholar
  8. Chandler, D.E. 1976. Stimulus-secretion coupling in the exocrine pancreas: Use of the fluorescent probe chlorotetracycline to detect divalent cation redistribution.Fed. Proc. 35:603 (Abstr.)Google Scholar
  9. Chandler, D.E., Williams, J.A. 1974. Pancreatic acinar cells: Effects of lanthanum ions on amylase release and calcium ion fluxes.J. Physiol. (London) 243:831Google Scholar
  10. Charles, M.A., Lawecki, J., Pictet, R., Grodsky, G.M. 1975. Insulin secretion. Interrelationships of glucose, cyclic adenosine 3′∶5′-monophosphate and calcium.J. Biol. Chem. 250:6134PubMedGoogle Scholar
  11. Cochrane, D.E., Douglas, W.W. 1974. Calcium induced extrusion of secretory granules exposed to 48/80 or the ionophores A23187 and X-537A.Proc. Nat. Acad. Sci. USA 71:408PubMedGoogle Scholar
  12. Cochrane, D.E., Douglas, W.W., Mouri, T., Nakazato, Y. 1975. Calcium and stimulus-secretion coupling in the adrenal medulla: Contrasting stimulating effects of the ionophores X-537A and A23187 on catecholamine output.J. Physiol. (London) 252:363Google Scholar
  13. Foreman, J.C., Mongar, J.L., Gomperts, B.D. 1973. Calcium ionophores and movement of calcium ions following the physiological stimulus to a secretory process.Nature (London) 245:249Google Scholar
  14. Garcia, A.G., Kirpekar, S.M., Prat, J.C. 1975. A calcium ionophore stimulating the secretion of catecholamines from the cat adrenal.J. Physiol. (London) 244:253Google Scholar
  15. Gardner, J.D., Conlon, T.P., Klaeveman, H.L., Adams, T.D., Ondetti, M.A. 1975. Action of cholecystokinin and cholingergic agents on calcium transport in isolated pancreatic acinar cells.J. Clin. Invest. 56:366PubMedGoogle Scholar
  16. Hainaut, K., Desmedt, J.E. 1974. Calcium ionophore A23187 potentiates twitch and intracellular calcium release in single muscle fibers.Nature (London) 252:407Google Scholar
  17. Heinz, E., Hoffman, J.F. 1965. Phosphate incorporation and Na, K-ATPase activity in human red blood cell ghosts.J. Cell Comp. Physiol. 65:31CrossRefGoogle Scholar
  18. Karl, R.C., Zawalich, W.S., Ferrendelli, J.A., Matschinsky, F.M. 1975. The role of Ca2+ and cyclic adenosine 3′∶5′-monophosphate in insulin release induced in vitro by the divalent cation ionophore A23187.J. Biol. Chem. 250:4575PubMedGoogle Scholar
  19. Kondo, S., Schulz, I. 1976. Calcium ion uptake in isolated pancreas cells induced by secretagogues.Biochim. Biophys. Acta. 419:76PubMedGoogle Scholar
  20. Murray, J.J., Reed, P.W., Fay, F.S. 1975. Contraction of isolated smooth muscle cells by ionophore A23187.Proc. Nat. Acad. Sci. USA 72:4459PubMedGoogle Scholar
  21. Petris, S. de 1975. Concanavalin A receptors, immunoglobulins, and θ-antigen of the lymphocyte surface.J. Cell Biol. 65:123CrossRefPubMedGoogle Scholar
  22. Pfeiffer, D.R., Reed, P.W., Lardy, H.A. 1974. Ultraviolet and fluorescent spectral properties of the divalent cation ionophore A23187 and its metal ion complexes.Biochemistry 13:4007PubMedGoogle Scholar
  23. Poulsen, J.H., Williams, J.A. 1976. Pancreatic acinar cell depolarization and amylase release evoked by Ca2+-ionophore A23187 and bethanechol.Fed. Proc. 35:603 (Abstr.)Google Scholar
  24. Pressman, B.C. 1972. Carboxylic ionophores as mobile carriers for divalent ions.In: The Role of Membranes in Metabolic Regulation M.A. Mehlman and R.W. Hanson, editors. p. 149. Academic Press, New YorkGoogle Scholar
  25. Reed, P.W., Lardy, H.A. 1972a. Antibiotic A23187 as a probe for the study of calcium and magnesium in biological systems.In: The Role of Membrane in Metabolic Regulation, A.M. Mehlman and R.W. Hanson, editors. p. 111. Academic Press, New YorkGoogle Scholar
  26. Reed, P.W., Lardy, H.A. 1972b. A23187: A divalent cation ionophore.J. Biol. Chem. 247:6970PubMedGoogle Scholar
  27. Rinderknecht, H., Wilding, P., Haverback, B.J. 1967. A new method for the determination of α-amylase.Experientia 23:805PubMedGoogle Scholar
  28. Robberecht, P., Christophe, J. 1971. Secretion of hydrolases by perfused fragments of rat pancreas: Effects of calcium.Am. J. Physiol. 220:911PubMedGoogle Scholar
  29. Russell, J.T., Hansen, E.L., Thorn, N.A. 1974. Calcium and stimulus-secretion coupling in the neurohypophysis. III. Ca2+ ionophore (A23187)-induced release of vasopressin from isolated rat neurohypophysis.Acta Endocrinol. (Copenhagen) 77:443Google Scholar
  30. Scarpa, A., Baldassare, J., Inesi, G. 1972. The effect of calcium ionophores on fragmented sarcoplasmic reticulum.J. Gen. Physiol. 60:735PubMedGoogle Scholar
  31. Schreurs, V.V.A.M., Swarts, H.G.P., dePont, J.J.H.M., Bonting, S.L. 1976. Role of calcium in exocrine pancreatic secretion. II. Comparison of the effects of carbachol and the ionophore A23187 on enzyme secretion and calcium movements in rabbit pancreas.Biochim. Biophys. Acta 419:320PubMedGoogle Scholar
  32. Schroeder, T.E., Strickland, D.L. 1974. Ionophore A23187, calcium and contractility in frog eggs.Exp. Cell Res. 83:139CrossRefPubMedGoogle Scholar
  33. Selinger, Z., Eimerl, S., Savion, N., Schramm, M. 1974. A Ca++ ionophore (A23187) simulating hormone and neurotransmitter action in the rat parotid and pancreas glands.In: Secretory Mechanisms of Exocrine Glands, N.A. Thorn and O.H. Petersen, editors. p. 68. Munksgaard, CopenhagenGoogle Scholar
  34. Sordahl, L.A. 1975. Effects of magnesium, ruthenium red and the antibiotic ionophore A23187 on initial rates of calcium uptake and release by heart mitochondria.Arch. Biochem. Biophys. 167:104CrossRefPubMedGoogle Scholar
  35. Steinhardt, R.A., Epel, D. 1974. Activation of sea-urchin eggs by a calcium ionophore.Proc. Nat. Acad. Sci. USA 71:1915PubMedGoogle Scholar
  36. Thoa, N.B., Costa, N.L., Moss, J., Kopin, I.J. 1974. Mechanism of release of norepinephrine from peripheral adrenergic neurones by the calcium ionophore X-537A and A23187.Life Sci. 14:1705PubMedGoogle Scholar
  37. Unanue, E.R., Perkins, W.D., Karnovsky, M.J. 1972. Ligand induced movement of lymphocyte membrane macromolecules. I. Analysis by immunofluorescence and ultrastructural radioautography.J. Exp. Med. 136:885CrossRefPubMedGoogle Scholar
  38. Wacker, W.E.C., Ulmer, D.D., Vallee, B.L. 1956. Metalloenzymes and myocardial infarction. II. Malic and latic dehydrogenase activities and zinc concentrations in serum.N. Engl. J. Med. 255:449Google Scholar
  39. Williams, J.A., Cary, P.C., Moffat, B. 1976. Effects of ions on amylase release by dissociated pancreatic acinar cells.Am. J. Physiol. 231:1562PubMedGoogle Scholar
  40. Williams, J.A., Chandler, D. 1975. Ca++ and pancreatic amylase release.Am. J. Physiol. 228:1729PubMedGoogle Scholar
  41. Williams, J.A., Lee, M. 1974. Pancreatic acinar cells: Use of a Ca++ ionophore to separate enzyme release from the earlier steps in stimulus-secretion coupling.Biochem. Biophys. Res. Commun. 60:542PubMedGoogle Scholar
  42. Wollheim, C.B., Blondel, B., Trueheart, P.A., Renold, A.E., Sharp, G.W.G. 1975. Calcium-induced insulin release in monolayer culture of the endocrine pancreas. Studies with ionophore A23187.J. Biol. Chem. 250:1354PubMedGoogle Scholar
  43. Wong, D.T., Wilkinson, J.R., Hamill, R.L., Horng, J.-S. 1973. Effects of antibiotic ionophore, A23187, on oxidative phosphorylation and calcium transport of liver mitochondria.Arch. Biochem. Biophys. 156:578CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag New York Inc 1977

Authors and Affiliations

  • Douglas E. Chandler
    • 1
  • John A. Williams
    • 1
  1. 1.Department of PhysiologyUniversity of CaliforniaSan Francisco

Personalised recommendations