Characterization of Purinergic Receptor-Evoked Increases in Intracellular Ca2+ Transients in Isolated Human and Rodent Insulin-Secreting Cells

Purinergic Receptor Signalling and [Ca2+]. in Human β-Cells
  • P. E. Squires
  • R. F. L. James
  • N. J. M. London
  • M. J. Dunne
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 426)


In vivo many factors determine the regulation of insulin secretion from the β-cells of the pancreatic islets of Langerhans. The most predominant influence is a rise in the concentration of plasma glucose, which will promote secretion by mechanisms associated with Ca2+ influx across the plasma membrane(1). Insulin release is also governed by changes in the concentration of circulating amino acids, gastrointestinal hormones, neuropeptides and neurotransmitters. These influences are important avenues for β-cell regulation as they not only provide a link between the gastrointestinal tract and the pancreatic islets — the enteroinsular axis, but they also govern the neurohormonal control of secretion. For many years it has been recognised that extracellular ATP is a potent insulin secretagogue(2). In vitro it has been demonstrated using rodent and clonal insulin-secreting cells that purinergic receptor activation is coupled to an increase in [Ca2+]i(3–6). However, few studies have examined intracellular Ca2+ signalling in isolated human insulin-secreting cells(4,7–10). In a recent paper we have shown that ATP and other agonists of the purinergic receptor evoke marked increases in [Ca2+]i in β-cells isolated from transplantable human islets of Langerhans(11). In this paper we examine: (i) the purinergic receptor subtype present in human tissue, (ii) compare purinergic receptor-evoked increases in [Ca2+]i in both rodent and human β-cells and (iii) demonstrate that intracellular Ca2+ signals can also be recorded in cryopreserved human β-cells originally isolated from cadaver organ donors.


RINm5F Cell United Kingdom Introduction Enteroinsular Axis Rodent Tissue Rodent Islet 
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. 1.
    Wollheim, C.B., and Sharp, G.W.G. 1981, Regulation of insulin release by calcium. Physiol Rev 61: 914–973.PubMedGoogle Scholar
  2. 2.
    Candela, J.L.R., and Garcia-Fernandez, M.C., 1963, Stimulation of secretion of insulin by adenosine triphosphate, Nature 197: 1210.CrossRefGoogle Scholar
  3. 3.
    Arkhammar, P., Hallberg, A., Kindmak, H., Nilsson, T., Rorsman. P., and Berggren, P.-O., 1990, Extracellular ATP increases cytoplasmic free Ca2+ concentration in clonal insulin-producing RINm5F cells, Biochem. J. 265:203–211.PubMedGoogle Scholar
  4. 4.
    Geschwind, J.-F., Hiriart, M, Glennon, M.C., Najafi, H., Corkey, B.E., Matschinsky, F.M., and Prentki, M., 1989, Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT), Biochim. Biophys. Acta 1012: 107–115.PubMedCrossRefGoogle Scholar
  5. 5.
    Hellman, B., Gylfe, E., Wesslen, N., Hallberg, A., Grapengiesser, E., and Marcstrom, A., 1989, Plasma membrane associated ATP as a regulator of the secretory activity of the pancreatic β-cell, Expt. Clinc. Endocrinol. 93: 125–135.CrossRefGoogle Scholar
  6. 6.
    Li, G., Milani, D., Dunne, M.J., Pralong, W.-F., Theler, J.-M., Petersen, O.H., and Wollheim, C.B., 1991, Extracellular ATP cause Ca2+-dependent and independent insulin secretion in RINm5F cells. J. Biol. Chem. 266: 3449–3457.PubMedGoogle Scholar
  7. 7.
    Sher, E., Biancardi, E., Pollo, A., Carbone, E., Li, G., Wollheim, C.B., and Clementi, F., 1992, Omega-Conotoxin-sensitive, voltage-operated calcium channels in insulin-secreting cells. Eur. J. Pharmacol. 216: 407–414.PubMedCrossRefGoogle Scholar
  8. 8.
    Pollo, A., Lovallo, M., Biancardi, E., Sher, E., Socci, C., and Carbone, E., 1993, Sensitivity to dihydropyridines, δ-conotoxin and noradrenaline reveals multiple high-voltage-activated calcium channels in rat insulinoma and human pancreatic β-cells, Pflugers Archiv. 423: 462–471.PubMedCrossRefGoogle Scholar
  9. 9.
    Kindmark, H., Kohler, M., Nillsson, T., Arkhammar, P., Wiechel, K.-L., Rorsman, P., Efandic, S., and Berggren, P.-O., 1992, Measurement of cytoplasmic free Ca2+ concentration in human pancreatic islets and insulinoma cells, FEBS Lett. 291: 310–314.CrossRefGoogle Scholar
  10. 10.
    Rojas, E., Caroli, P.B., Ricordi, C., Boschero, A.C., Stojilikovic, S.S., and Atwater, I., 1994, Control of cytosolic free calcium in cultured human pancreatic β-cells occurs by external calcium dependent and independent mechanisms, Endocrinology 134: 1771–1781.PubMedCrossRefGoogle Scholar
  11. 11.
    Squires, P.E., James, R.F.L., London, N.J.M., and Dunne, M.J., 1994, ATP-induced intracellular Ca2+ signals in isolated human insulin-secreting cells, Pflugers Archives 427: 181–183.CrossRefGoogle Scholar
  12. 12.
    London, N.J.M., James, R.F.L., and Bell, P.R.F., 1992, Islet purification. In: Pancreatic Islet Cell Transplantation, pp 113-123, Edited by C. Ricordi, Landes Press.Google Scholar
  13. 13.
    Rich, S.J., Swift, S., Thirdborough, S.M., Rumford, G., James, R.F.L., and London, N.J.M., 1993, Cryopreservation of rat islets of Langerhans: a comparison of two techniques, Cryobiology 30: 407–412.PubMedCrossRefGoogle Scholar
  14. 14.
    Findlay, I., Dunne, MJ. and Petersen, O.H., 1985, High conductance K+ channel in pancreatic islet cells can be activated and inactivated by internal calcium, J. Memb. Biol. 83: 169–175.CrossRefGoogle Scholar
  15. 15.
    Dubyak, G.R., and El-Moatassim, C., 1993, Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides, Am. J. Physiol. 265: C577–C606.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1997

Authors and Affiliations

  • P. E. Squires
    • 1
  • R. F. L. James
    • 2
  • N. J. M. London
    • 2
  • M. J. Dunne
    • 1
  1. 1.Department of Biomedicai ScienceThe University of SheffieldSheffieldUK
  2. 2.Department of Surgery, School of MedicineThe University of LeicesterLeicesterUK

Personalised recommendations