Diabetologia

, Volume 37, Issue 10, pp 1007–1014 | Cite as

Culture duration and conditions affect the oscillations of cytoplasmic calcium concentration induced by glucose in mouse pancreatic islets

  • P. Gilon
  • J. C. Jonas
  • J. C. Henquin
Originals

Summary

The pattern of the increase in cytoplasmic Cai2+ that glucose produces in beta cells has been reported to be highly variable. Here, we evaluated the influence of the culture duration (1–4 days) and conditions (5–10 mmol/l glucose) on Cai2+ in normal mouse islets stimulated by glucose. After 1 day of culture in 10 mmol/l glucose, a rise of the glucose concentration from 3 to 15 mmol/l induced a triphasic change of Cai2+ in the islets. A small initial decrease was followed by a large peak increase and then by regular fast oscillations (∼2.5/min). When the culture was prolonged to 2, 3 and 4 days, the initial decrease became inconsistent and the peak occurred earlier, whereas the oscillations decreased in frequency, increased in duration and eventually disappeared; on day 4 the Cai2+ rise was sustained. After culture in 5 mmol/l glucose, the pattern of Cai2+ changes induced by 15 mmol/l glucose was different. The initial decrease was very pronounced, the first peak was delayed and clearly separated from the subsequent oscillations. These were of a mixed type (fast Ca2+ transients on top of slow ones) after 1 day, and of a slow type only after 4 days. These alterations in the Cai2+ oscillations triggered by glucose could not be ascribed to desynchronization of the signal between different regions of the islets. In conclusion, culturing normal mouse islets in 5 or 10 mmol/l glucose for 1–4 days, markedly alters the characteristics of the changes in Cai2+ produced by glucose. This pitfall must be borne in mind when studying stimulus-secretion coupling in beta cells from normal or diabetic animals, or from human islets.

Key words

Islets culture calcium glucose stimulus-secretion coupling beta cells 

Abbreviation

Cai2+

concentration of cytoplasmic calcium

References

  1. 1.
    Grodsky GM, Bennett LL (1966) Cation requirements for insulin secretion in the isolated perfused pancreas. Diabetes 15: 910–913Google Scholar
  2. 2.
    Milner RDG, Hales CN (1967) The role of calcium and magnesium in insulin secretion from rabbit pancreas studied in vitro. Diabetologia 3: 47–49Google Scholar
  3. 3.
    Wollheim CB, Sharp GWG (1981) The regulation of insulin release by calcium. Physiol Rev 61: 914–973Google Scholar
  4. 4.
    Henquin JC, Meissner HP (1984) Significance of ionic fluxes and changes in membrane potential for stimulus-secretion coupling in pancreatic B-cells. Experientia 40: 1043–1052Google Scholar
  5. 5.
    Prentki M, Matschinsky FM (1987) Ca2+, cAMP, and phospholipid-derived messengers in coupling mechanisms of insulin secretion. Physiol Rev 67: 1185–1248Google Scholar
  6. 6.
    Ashcroft FM, Rorsman P (1989) Electrophysiology of the pancreatic Β-cell. Prog Biophys Mol Biol 54: 87–143Google Scholar
  7. 7.
    Cook DL, Satin LS, Hopkins WF (1991) Pancreatic Β cells are bursting, but how. Trends Neurosci. 14: 411–414Google Scholar
  8. 8.
    Rorsman P, Abrahamsson H, Gylfe E, Hellman B (1984) Dual effects of glucose on the cytosolic Ca2+ activity of mouse pancreatic Β-cells. FEBS Lett 170: 196–200Google Scholar
  9. 9.
    Deleers M, Mahy M, Malaisse WJ (1985) Glucose increases cytosolic Ca2+ activity in pancreatic islet cells. Biochem Int 10: 97–103Google Scholar
  10. 10.
    Wollheim CB, Pozzan T (1984) Correlation between cytosolic free Ca2+ and insulin release in an insulin-secreting cell line. J Biol Chem 25: 2262–2267Google Scholar
  11. 11.
    Herchuelz A, Juvent M, Van Ganse E, Gobbe P (1986) Differential effect of nutrient and non-nutrient secretagogues on cytosolic free Ca2+ in pancreatic islet cells. In: Atwater I, Rojas E, Soria B (eds) Biophysics of the pancreatic Β-cell. New York, Plenum Press, pp 317–318Google Scholar
  12. 12.
    Sussman KE, Leitner JW, Draznin B (1987) Cytosolic free-calcium concentrations in normal pancreatic islet cells. Effect of secretagogues and somatostatin. Diabetes 36: 571–577Google Scholar
  13. 13.
    Gylfe E (1988) Glucose-induced early changes in cytoplasmic calcium of pancreatic Β-cells studied with time-sharing dual-wavelength fluorometry. J Biol Chem 263: 5044–5048Google Scholar
  14. 14.
    Grapengiesser E, Gylfe E, Hellman B (1988) Glucose-induced oscillations of cytoplasmic Ca2+ in the pancreatic Β-cell. Biochem Biophys Res Commun 151: 1299–1304Google Scholar
  15. 15.
    Wang J-L, McDaniel ML (1990) Secretagogue-induced oscillations of cytoplasmic Ca2+ in single Β and α-cells obtained from pancreatic islets by fluorescence-activated cell sorting. Biochem Biophys Res Commun 166: 813–818Google Scholar
  16. 16.
    Pralong W-F, Bartley C, Wollheim CB (1990) Single islet Β-cell stimulation by nutrients: relationship between pyridine nucleotides, cytosolic Ca2+ and secretion. EMBO J 9: 53–60Google Scholar
  17. 17.
    Herchuelz A, Pochet R, Pastiels Ch, Van Praet A (1991) Heterogeneous changes in [Ca2+]i induced by glucose, tolbutamide and K+ in single rat pancreatic B cells. Cell Calcium 12: 577–586Google Scholar
  18. 18.
    Hellman B, Gylfe E, Grapengiesser E, Lund P-E, Berts A (1992) Cytoplasmic Ca2+ oscillations in pancreatic Β-cells. Biochim Biophys Acta 1113: 295–305Google Scholar
  19. 19.
    Theler JM, Mollard P, Guérineau N, Vacher P, Pralong WF, Schlegel W, Wollheim CB (1992) Video imaging of cytosolic Ca2+ in pancreatic Β-cells stimulated by glucose, carbachol, and ATP. J Biol Chem 267: 18110–18117Google Scholar
  20. 20.
    Wang JL, Corbett JA, Marshall CA, McDaniel ML (1993) Glucose-induced insulin secretion from purified Β-cells. A role for modulation of Ca2+ influx by cAMP- and protein kinase C-dependent signal transduction pathways. J Biol Chem 268: 7785–7791Google Scholar
  21. 21.
    Valdeolmillos M, Santos RM, Contreras D, Soria B, Rosario LM (1989) Glucose-induced oscillations of intracellular Ca2+ concentration resembling bursting electrical activity in single mouse islets of Langerhans. FEBS Lett 25: 19–23Google Scholar
  22. 22.
    Santos RM, Rosario LM, Nadal A, Garcia-Sancho J, Soria B, Valdeolmillos M (1991) Widespread synchronous [Ca2+]i oscillations due to bursting electrical activity in single pancreatic islets. Pflügers Arch 418: 417–422Google Scholar
  23. 23.
    Zhang A, Gao ZY, Gilon P, Nenquin M, Drews G, Henquin JC (1991) Vanadate stimulation of insulin release in normal mouse islets. J Biol Chem 266: 21649–21656Google Scholar
  24. 24.
    Gilon P, Henquin JC (1992) Influence of membrane potential changes on cytoplasmic Ca2+ concentration in an electrically excitable cell, the insulin-secreting pancreatic B-cell. J Biol Chem 267: 20713–20720Google Scholar
  25. 25.
    Longo EA, Tornheim K, Deeney JT, Varnum BA, Tillotson D, Prentki M, Corkey BE (1991) Oscillations in cytosolic free Ca2+, oxygen consumption, and insulin secretion in glucose-stimulated rat pancreatic islets. J Biol Chem 266: 9314–9319Google Scholar
  26. 26.
    Gilon P, Shepherd RM, Henquin JC (1993) Oscillations of secretion driven by oscillations of cytoplasmic Ca2+ as evidenced in single pancreatic islets. J Biol Chem 268: 22265–22268Google Scholar
  27. 27.
    Yada T, Kakei M, Tanaka H (1992) Single pancreatic Β-cells from normal rats exhibit an initial decrease and subsequent increase in cytosolic free Ca2+ in response to glucose. Cell Calcium 13: 69–76Google Scholar
  28. 28.
    Smith PA, Ashcroft FM, Rorsman P (1990) Simultaneous recordings of glucose dependent electrical activity and ATP-regulated K+-currents in isolated mouse pancreatic Β-cells. FEBS Lett 261: 187–190Google Scholar
  29. 29.
    Dunne MJ, Petersen OH (1991) Potassium selective ion channels in insulin-secreting cells: physiology, pharmacology and their role in stimulus-secretion coupling. Biochim Biophys Acta 1071: 67–82Google Scholar
  30. 30.
    Smolen P, Rinzel J, Sherman A (1993) Why pancreatic islets burst but single Β cells do not. The heterogeneity hypothesis. Biophys J 64: 1668–1680Google Scholar
  31. 31.
    Andersson A (1974) Long-term effects of glucose on insulin release and glucose oxidation by mouse pancreatic islets maintained in tissue culture. Biochem J 140: 377–382Google Scholar
  32. 32.
    Svensson C, Hellerström C (1991) Long-term effects of a high glucose concentration in vitro on the oxidative metabolism and insulin production of isolated rat pancreatic islets. Metabolism 40: 513–518Google Scholar
  33. 33.
    Henquin JC (1992) The biophysical events involved in the stimulation of insulin release by arginine. In: De Deyn PP, Marescau B, Stalon V, Qureshi IA (eds) Guanidino compounds in biology and medicine. John Libbey & Company Ltd, pp 109–116Google Scholar
  34. 34.
    Bergsten P, Grapengiesser E, Gylfe E, Tengholm A, Hellman B (1994) Synchronous oscillations of cytoplasmic Ca2+ and insulin release in glucose-stimulated pancreatic islets. J Biol Chem 269: 8749–8753Google Scholar
  35. 35.
    Henquin JC, Meissner HP, Schmeer W (1982) Cyclic variations of glucose-induced electrical activity in pancreatic B-cells. Pflügers Arch 393: 322–327Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • P. Gilon
    • 1
  • J. C. Jonas
    • 1
  • J. C. Henquin
    • 1
  1. 1.Unité d'Endocrinologie et MétabolismeUniversity of Louvain Faculty of MedicineBrusselsBelgium

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