Skip to main content
SpringerLink
Log in

Capacitative Ca2+ influx and a Ca2u+-dependent nonselective cation pathway are discriminated by genistein in mouse pancreatic acinar cells

  • Original Article
  • Molecular and Cellular Physiology
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

We have investigated the effect of genistein on the hormone-stimulated Ca2+ influx and on a 28 pS nonselective cation channel in mouse pancreatic acinar cells using the Ca2+ indicator fluo-3 and the patch-clamp technique. The identity of the Ca2u+ influx pathway has not been established in this cell type so far. Therefore we have investigated the Ca2+-dependent nonselective cation channel as a potential pathway for Ca2+ influx. “Capacitative” Ca2+ entry was induced by depletion of intracellular Ca2+ stores with 500 nM acetylcholine or with the Ca2+ ATPase inhibitor 2,5-di-tert- butylhydroquinone. In the presence of 100 μM genistein, Ca2+ release was unimpaired, whereas Ca2+ influx was reversibly suppressed. Patch-clamp experiments demonstrated that genistein had no effect on Ca2+-activated nonselective cation channels, the activity of which was measured in excised membrane patches (inside/out) or in the whole-cell configuration. Therefore we conclude that this 28 pS nonselective cation channel does not contribute to Ca2+ influx into mouse exocrine pancreatic cells. With the exception of genistein and tyrphostin 25, other tyrosine kinase inhibitors such as methyl-2,5-dihydroxycinnamate, lavendustin A, herbimycin A, and tyrphostin B56 were without effect on Ca2+ signalling. Thus, the involvement of tyrosine phosphorylation in the activation of the Ca2+ entry mechanism in mouse pancreatic acinar cells is unclear.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Akiyama T, Ishida J, Nakagawa S, Ogawara H, Watanabe S, Itoh N, Shibuya M, Fukami Y (1987) Genistein, a specific inhibitor of tyrosine-specific protein kinases. J Biol Chem 262:5592–5595

    PubMed  Google Scholar 

  2. Bahnson TD, Pandol SJ, Dionne VE (1993) Cyclic GMP modulates depletion-activated Ca2+ entry in pancreatic acinar cells. J Biol Chem 268:10808–10812

    PubMed  Google Scholar 

  3. Bird G StJ, Putney JW (1993) Inhibition of thapsigargin-induced calcium entry by microinjected guanine nucleotide analogues. J Biol Chem 268:21486–21488

    PubMed  Google Scholar 

  4. Bischof G, Illek B, Reenstra WW, Machen E (1995) Role of tyrosine kinases in carbachol-regulated Ca entry into colonic epithelial cells. Am J Physiol 268:C154-C161

    PubMed  Google Scholar 

  5. Duan R, Wagner ACC, Yule DI, Williams JA (1994) Multiple inhibitory effects of genistein on stimulus-secretion coupling in rat pancreatic acini. Am J Physiol 266:G303-G310

    PubMed  Google Scholar 

  6. Fasolato C, Hoth M, Penner R (1993) A GTP-dependent step in the activation mechanism of capacitative calcium influx. J Biol Chem 268:20737–20740

    PubMed  Google Scholar 

  7. Gamberucci A, Innocenti B, Fulceri R, Bànhegyi G, Giunti R, Pozzan T, Benedetti A (1994) Modulation of Ca2+influx dependent on store depletion by intracellular adenine-guanine nucleotide levels. J Biol Chem 269:23597–23602

    PubMed  Google Scholar 

  8. Gukovskaya A, Pandol S (1994) Nitric oxide production regulates cGMP formation and calcium influx in pancreatic acinar cells. Am J Physiol 266:G350-G356

    PubMed  Google Scholar 

  9. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100

    Article  Google Scholar 

  10. Hidaka H, Inagaki M, Kawamoto S, Sasaki Y (1984) Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 23:5036–5041

    PubMed  Google Scholar 

  11. Hoth M, Penner R (1992) Depletion of intracellular calcium stores activates a calcium current in mast cells. Nature 355:353–356

    PubMed  Google Scholar 

  12. Irvine RF (1992) Inositol phosphates and Ca2+ entry: toward a proliferation or a simplification? FASEB J 6: 3085–3091

    PubMed  Google Scholar 

  13. Kondo S, Schulz I (1976) Ca2+ uptake in isolated pancreas cells induced by secretagogues. Biochim Biophys Acta 419: 76–92

    PubMed  Google Scholar 

  14. Lee K, Toscas K, Villereal ML (1993) Inhibition of bradykinin- and thapsigargin-induced Ca2+ entry by tyrosine kinase inhibitors. J Biol Chem 268:9945–9948

    PubMed  Google Scholar 

  15. Lutz MP, Sutor SL, Abraham RT, Miller L (1993) A role for cholecystokinin-stimulated protein tyrosine phosphorylation in regulated secretion by the pancreatic acinar cell. J Biol Chem 268:11119–11124

    PubMed  Google Scholar 

  16. Maruyama Y, Petersen OH (1982) Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini. Nature 299:159–161

    PubMed  Google Scholar 

  17. Montero M, Alvarez J, García-Sancho J (1992) Control of plasma-membrane Ca2+ entry by the intracellular Ca2+ stores. Biochem J 288:519–525

    PubMed  Google Scholar 

  18. Moore GA, McConkey DJ, Kass GEN, O'Brien PJ, Orrenius S (1987) 2,5-Di(tert-butyl)-1,4-benzohydroquinonea novel inhibitor of liver microsomal Ca2+ sequestration. FEBS Lett 224:331–336

    PubMed  Google Scholar 

  19. Nilius B (1990) Permeation properties of a non-selective cation channel in human vascular endothelial cells. Pflügers Arch 416:609–611

    Article  Google Scholar 

  20. Osipchuk YV, Wakui M, Yule DI, Gallacher DV, Petersen OH (1990) Cytoplasmic Ca2+ oscillations evoked by receptor stimulation, G-protein activation, internal application of inositol trisphosphate or Ca2+: simultaneous microfluorimetry and Ca2+ dependent Cl− current recording in single pancreatic acinar cells. EMBO J 9:697–704

    PubMed  Google Scholar 

  21. Ozaki Y, Yatomi Y, Jinnai Y, Kume S (1993) Effects of genistein, a tyrosine kinase inhibitor, on platelet functions. Biochem Pharmacol 46:395–403

    PubMed  Google Scholar 

  22. Parekh AB, Terlau H, Stühmer W (1994) Depletion of InsP3 stores activates a Ca2+ and K+ current by means of a phosphatase and a diffusible messenger. Nature 364:814–818

    Article  Google Scholar 

  23. Penner R, Matthews G, Neher E (1988) Regulation of calcium influx by second messengers in rat mast cells. Nature 344:499–504

    Article  Google Scholar 

  24. Petersen OH (1992) Stimulus-secretion coupling: cytoplasmic calcium signals and the control of ion channels in exocrine acinar cells. J Physiol (Lond) 448:1–51

    Google Scholar 

  25. Petersen OH, Maruyama Y (1983) What is the mechanism of the calcium influx to pancreatic acinar cells evoked by secretagogues? Pflügers Arch 396:82–84

    Article  Google Scholar 

  26. Piiper A, Stryjek-Kaminska D, Stein J, Caspary WF, Zeuzem S (1994) Tyrphostins inhibit secretagogue-induced 1,4,5-IP3production and amylase release in pancreatic acini. Am J Physiol 266:G363-G371

    PubMed  Google Scholar 

  27. Poronnik P, Cook DI, Allen DG, Young JA (1991) Diphenylamine-2-carboxylate (DPC) reduces calcium influx in a mouse madibular cell line (ST885). Cell Calcium 12:441–447

    PubMed  Google Scholar 

  28. Putney JW Jr, Bird G StJ (1993) The signal for capacitative calcium entry. Cell 75:199–201

    PubMed  Google Scholar 

  29. Randriamampita C, Tsien RY (1994) Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature 364:809–814

    Article  Google Scholar 

  30. Sargeant P, Farndale RW, Sage SO (1993) ADP- and thapsigargin-evoked Ca2+ entry and protein-tyrosine phosphorlylation are inhibited by the tyrosine kinase inhibitors genistein and methyl-2,5-dihydroxycinnamate in fura-2 loaded human platelets. J Biol Chem 268:18151–18156

    PubMed  Google Scholar 

  31. Schulz I (1980) Messenger role of calcium in function of pancreatic acinar cells. Am J Physiol 239:G335-G347

    PubMed  Google Scholar 

  32. Streb H, Schulz I (1984) Regulation of cytosolic free Ca2+ concentration in acinar cells of rat pancreas. Am J Physiol 245:G347-G357

    Google Scholar 

  33. Tepel M, Kühnapfel S, Theilmeier G, Teupe C, Schlotmann R, Zidek W (1994) Filling state of intracellular Ca2+ pools triggers trans plasma membrane Na+ and Ca2+ influx by a tyrosine kinase-dependent pathway. J Biol Chem 269:26239–26242

    PubMed  Google Scholar 

  34. Thastrup O, Dawson AP, Scharff O, Foder B, Bjenum PJ, Hanley MJ (1989) Thapsigargin, a novel molecular probe for studying intracellular calcium release and storage. Agents Actions 27:17–23

    PubMed  Google Scholar 

  35. Thorn P, Petersen OH (1993) Nonselective cation channels in exocrine gland cells. In: Siemen D, Hescheler J (eds) Nonselective cation channels: pharmacology, physiology and biophysics. Birkhäuser Verlag, Basel, pp 185–200

    Google Scholar 

  36. Tscharner V, Prod'hom B, Baggiolini M, Reuter H (1986) Ion channels in human neutrophils activated by a rise in free cytosolic calcium concentration. Nature 324:369–372

    PubMed  Google Scholar 

  37. Vostal JG, Jackson WL, Shulman NR (1991) Cytosolic and stored calcium antagonistically control tyrosine phosphorylation of specific platelet proteins. J Biol Chem 266: 16911–16916

    PubMed  Google Scholar 

  38. Xu X, Star RA, Tortorici G, Muallem S (1994) Depletion of intracellular Ca2+ stores activates nitric-oxide synthase to generate cGMP and regulate Ca2+influx. J Biol Chem 269: 12645–12653

    PubMed  Google Scholar 

  39. Yule DI, Kim ET, Williams JA (1994) Tyrosine kinase inhibitors attenuate “capacitative” Ca2+ influx in rat pancreatic acinar cells. Bichem Biophys Res Commun 202:1697–1704

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. 2. Physiologisches Institut, Universität des Saarlandes, D-66421, Homburg/Saar, Germany

    Fatima Pfeiffer, Andreas Schmid & Irene Schulz

Authors
  1. Fatima Pfeiffer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andreas Schmid
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Irene Schulz
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pfeiffer, F., Schmid, A. & Schulz, I. Capacitative Ca2+ influx and a Ca2u+-dependent nonselective cation pathway are discriminated by genistein in mouse pancreatic acinar cells. Pflugers Arch. 430, 916–922 (1995). https://doi.org/10.1007/BF01837405

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01837405

Key words

Search

Navigation