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. 1998 Jan 15;17(2):435–442. doi: 10.1093/emboj/17.2.435

Termination of cytosolic Ca2+ signals: Ca2+ reuptake into intracellular stores is regulated by the free Ca2+ concentration in the store lumen.

H Mogami 1, A V Tepikin 1, O H Petersen 1
PMCID: PMC1170394  PMID: 9430635

Abstract

The mechanism by which agonist-evoked cytosolic Ca2+ signals are terminated has been investigated. We measured the Ca2+ concentration inside the endoplasmic reticulum store of pancreatic acinar cells and monitored the cytoplasmic Ca2+ concentration by whole-cell patch-clamp recording of the Ca2+-sensitive currents. When the cytosolic Ca2+ concentration was clamped at the resting level by a high concentration of a selective Ca2+ buffer, acetylcholine evoked the usual depletion of intracellular Ca2+ stores, but without increasing the Ca2+-sensitive currents. Removal of acetylcholine allowed thapsigargin-sensitive Ca2+ reuptake into the stores, and this process stopped when the stores had been loaded to the pre-stimulation level. The apparent rate of Ca2+ reuptake decreased steeply with an increase in the Ca2+ concentration in the store lumen and it is this negative feedback on the Ca2+ pump that controls the Ca2+ store content. In the absence of a cytoplasmic Ca2+ clamp, acetylcholine removal resulted in a rapid return of the elevated cytoplasmic Ca2+ concentration to the pre-stimulation resting level, which was attained long before the endoplasmic reticulum Ca2+ store had been completely refilled. We conclude that control of Ca2+ reuptake by the Ca2+ concentration inside the intracellular store allows precise Ca2+ signal termination without interfering with store refilling.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belan P. V., Gerasimenko O. V., Tepikin A. V., Petersen O. H. Localization of Ca2+ extrusion sites in pancreatic acinar cells. J Biol Chem. 1996 Mar 29;271(13):7615–7619. doi: 10.1074/jbc.271.13.7615. [DOI] [PubMed] [Google Scholar]
  2. Belan P., Gerasimenko O., Petersen O. H., Tepikin A. V. Distribution of Ca2+ extrusion sites on the mouse pancreatic acinar cell surface. Cell Calcium. 1997 Jul;22(1):5–10. doi: 10.1016/s0143-4160(97)90084-1. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J. Elementary and global aspects of calcium signalling. J Physiol. 1997 Mar 1;499(Pt 2):291–306. doi: 10.1113/jphysiol.1997.sp021927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J. Inositol trisphosphate and calcium signalling. Nature. 1993 Jan 28;361(6410):315–325. doi: 10.1038/361315a0. [DOI] [PubMed] [Google Scholar]
  5. Bezprozvanny I., Watras J., Ehrlich B. E. Bell-shaped calcium-response curves of Ins(1,4,5)P3- and calcium-gated channels from endoplasmic reticulum of cerebellum. Nature. 1991 Jun 27;351(6329):751–754. doi: 10.1038/351751a0. [DOI] [PubMed] [Google Scholar]
  6. Bootman M. D., Berridge M. J. The elemental principles of calcium signaling. Cell. 1995 Dec 1;83(5):675–678. doi: 10.1016/0092-8674(95)90179-5. [DOI] [PubMed] [Google Scholar]
  7. Camello P., Gardner J., Petersen O. H., Tepikin A. V. Calcium dependence of calcium extrusion and calcium uptake in mouse pancreatic acinar cells. J Physiol. 1996 Feb 1;490(Pt 3):585–593. doi: 10.1113/jphysiol.1996.sp021169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clapham D. E. Calcium signaling. Cell. 1995 Jan 27;80(2):259–268. doi: 10.1016/0092-8674(95)90408-5. [DOI] [PubMed] [Google Scholar]
  9. Ebashi S., Lipmann F. ADENOSINE TRIPHOSPHATE-LINKED CONCENTRATION OF CALCIUM IONS IN A PARTICULATE FRACTION OF RABBIT MUSCLE. J Cell Biol. 1962 Sep 1;14(3):389–400. doi: 10.1083/jcb.14.3.389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Favre C. J., Schrenzel J., Jacquet J., Lew D. P., Krause K. H. Highly supralinear feedback inhibition of Ca2+ uptake by the Ca2+ load of intracellular stores. J Biol Chem. 1996 Jun 21;271(25):14925–14930. doi: 10.1074/jbc.271.25.14925. [DOI] [PubMed] [Google Scholar]
  11. Gerasimenko O. V., Gerasimenko J. V., Belan P. V., Petersen O. H. Inositol trisphosphate and cyclic ADP-ribose-mediated release of Ca2+ from single isolated pancreatic zymogen granules. Cell. 1996 Feb 9;84(3):473–480. doi: 10.1016/s0092-8674(00)81292-1. [DOI] [PubMed] [Google Scholar]
  12. Hajnóczky G., Thomas A. P. Minimal requirements for calcium oscillations driven by the IP3 receptor. EMBO J. 1997 Jun 16;16(12):3533–3543. doi: 10.1093/emboj/16.12.3533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  14. Hofer A. M., Machen T. E. Technique for in situ measurement of calcium in intracellular inositol 1,4,5-trisphosphate-sensitive stores using the fluorescent indicator mag-fura-2. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2598–2602. doi: 10.1073/pnas.90.7.2598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hofer A. M., Schlue W. R., Curci S., Machen T. E. Spatial distribution and quantitation of free luminal [Ca] within the InsP3-sensitive internal store of individual BHK-21 cells: ion dependence of InsP3-induced Ca release and reloading. FASEB J. 1995 Jun;9(9):788–798. doi: 10.1096/fasebj.9.9.7601343. [DOI] [PubMed] [Google Scholar]
  16. Hofer A. M., Schulz I. Quantification of intraluminal free [Ca] in the agonist-sensitive internal calcium store using compartmentalized fluorescent indicators: some considerations. Cell Calcium. 1996 Sep;20(3):235–242. doi: 10.1016/s0143-4160(96)90029-9. [DOI] [PubMed] [Google Scholar]
  17. Inesi G., de Meis L. Regulation of steady state filling in sarcoplasmic reticulum. Roles of back-inhibition, leakage, and slippage of the calcium pump. J Biol Chem. 1989 Apr 5;264(10):5929–5936. [PubMed] [Google Scholar]
  18. KUMAGAI H., EBASHI S., TAKEDA F. Essential relaxing factor in muscle other than myokinase and creatine phosphokinase. Nature. 1955 Jul 23;176(4473):166–166. doi: 10.1038/176166a0. [DOI] [PubMed] [Google Scholar]
  19. Kasai H., Augustine G. J. Cytosolic Ca2+ gradients triggering unidirectional fluid secretion from exocrine pancreas. Nature. 1990 Dec 20;348(6303):735–738. doi: 10.1038/348735a0. [DOI] [PubMed] [Google Scholar]
  20. Kasai H., Li Y. X., Miyashita Y. Subcellular distribution of Ca2+ release channels underlying Ca2+ waves and oscillations in exocrine pancreas. Cell. 1993 Aug 27;74(4):669–677. doi: 10.1016/0092-8674(93)90514-q. [DOI] [PubMed] [Google Scholar]
  21. Lee M. G., Xu X., Zeng W., Diaz J., Kuo T. H., Wuytack F., Racymaekers L., Muallem S. Polarized expression of Ca2+ pumps in pancreatic and salivary gland cells. Role in initiation and propagation of [Ca2+]i waves. J Biol Chem. 1997 Jun 20;272(25):15771–15776. doi: 10.1074/jbc.272.25.15771. [DOI] [PubMed] [Google Scholar]
  22. Maruyama Y., Petersen O. H. Cholecystokinin activation of single-channel currents is mediated by internal messenger in pancreatic acinar cells. Nature. 1982 Nov 4;300(5887):61–63. doi: 10.1038/300061a0. [DOI] [PubMed] [Google Scholar]
  23. Missiaen L., De Smedt H., Droogmans G., Casteels R. 2,5-Di-(tert-butyl)-1,4-benzohydroquinone and cyclopiazonic acid decrease the Ca2+ permeability of endoplasmic reticulum. Eur J Pharmacol. 1992 Dec 1;227(4):391–394. doi: 10.1016/0922-4106(92)90156-p. [DOI] [PubMed] [Google Scholar]
  24. Mogami H., Nakano K., Tepikin A. V., Petersen O. H. Ca2+ flow via tunnels in polarized cells: recharging of apical Ca2+ stores by focal Ca2+ entry through basal membrane patch. Cell. 1997 Jan 10;88(1):49–55. doi: 10.1016/s0092-8674(00)81857-7. [DOI] [PubMed] [Google Scholar]
  25. Montero M., Barrero M. J., Alvarez J. [Ca2+] microdomains control agonist-induced Ca2+ release in intact HeLa cells. FASEB J. 1997 Sep;11(11):881–885. doi: 10.1096/fasebj.11.11.9285486. [DOI] [PubMed] [Google Scholar]
  26. Montero M., Brini M., Marsault R., Alvarez J., Sitia R., Pozzan T., Rizzuto R. Monitoring dynamic changes in free Ca2+ concentration in the endoplasmic reticulum of intact cells. EMBO J. 1995 Nov 15;14(22):5467–5475. doi: 10.1002/j.1460-2075.1995.tb00233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Osipchuk Y. V., Wakui M., Yule D. I., Gallacher D. V., Petersen O. H. 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. 1990 Mar;9(3):697–704. doi: 10.1002/j.1460-2075.1990.tb08162.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Petersen C. C., Petersen O. H., Berridge M. J. The role of endoplasmic reticulum calcium pumps during cytosolic calcium spiking in pancreatic acinar cells. J Biol Chem. 1993 Oct 25;268(30):22262–22264. [PubMed] [Google Scholar]
  29. Petersen O. H., Petersen C. C., Kasai H. Calcium and hormone action. Annu Rev Physiol. 1994;56:297–319. doi: 10.1146/annurev.ph.56.030194.001501. [DOI] [PubMed] [Google Scholar]
  30. Petersen O. H. Stimulus-secretion coupling: cytoplasmic calcium signals and the control of ion channels in exocrine acinar cells. J Physiol. 1992 Mar;448:1–51. doi: 10.1113/jphysiol.1992.sp019028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pozzan T., Rizzuto R., Volpe P., Meldolesi J. Molecular and cellular physiology of intracellular calcium stores. Physiol Rev. 1994 Jul;74(3):595–636. doi: 10.1152/physrev.1994.74.3.595. [DOI] [PubMed] [Google Scholar]
  32. Putney J. W., Jr Capacitative calcium entry revisited. Cell Calcium. 1990 Nov-Dec;11(10):611–624. doi: 10.1016/0143-4160(90)90016-n. [DOI] [PubMed] [Google Scholar]
  33. Renard-Rooney D. C., Hajnóczky G., Seitz M. B., Schneider T. G., Thomas A. P. Imaging of inositol 1,4,5-trisphosphate-induced Ca2+ fluxes in single permeabilized hepatocytes. Demonstration of both quantal and nonquantal patterns of Ca2+ release. J Biol Chem. 1993 Nov 5;268(31):23601–23610. [PubMed] [Google Scholar]
  34. Roberts W. M. Spatial calcium buffering in saccular hair cells. Nature. 1993 May 6;363(6424):74–76. doi: 10.1038/363074a0. [DOI] [PubMed] [Google Scholar]
  35. Sagara Y., Wade J. B., Inesi G. A conformational mechanism for formation of a dead-end complex by the sarcoplasmic reticulum ATPase with thapsigargin. J Biol Chem. 1992 Jan 15;267(2):1286–1292. [PubMed] [Google Scholar]
  36. Simon S. M., Blobel G. A protein-conducting channel in the endoplasmic reticulum. Cell. 1991 May 3;65(3):371–380. doi: 10.1016/0092-8674(91)90455-8. [DOI] [PubMed] [Google Scholar]
  37. Subramanian K., Meyer T. Calcium-induced restructuring of nuclear envelope and endoplasmic reticulum calcium stores. Cell. 1997 Jun 13;89(6):963–971. doi: 10.1016/s0092-8674(00)80281-0. [DOI] [PubMed] [Google Scholar]
  38. Tepikin A. V., Voronina S. G., Gallacher D. V., Petersen O. H. Acetylcholine-evoked increase in the cytoplasmic Ca2+ concentration and Ca2+ extrusion measured simultaneously in single mouse pancreatic acinar cells. J Biol Chem. 1992 Feb 25;267(6):3569–3572. [PubMed] [Google Scholar]
  39. Tepikin A. V., Voronina S. G., Gallacher D. V., Petersen O. H. Pulsatile Ca2+ extrusion from single pancreatic acinar cells during receptor-activated cytosolic Ca2+ spiking. J Biol Chem. 1992 Jul 15;267(20):14073–14076. [PubMed] [Google Scholar]
  40. Thastrup O., Cullen P. J., Drøbak B. K., Hanley M. R., Dawson A. P. Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2(+)-ATPase. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2466–2470. doi: 10.1073/pnas.87.7.2466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thorn P., Lawrie A. M., Smith P. M., Gallacher D. V., Petersen O. H. Local and global cytosolic Ca2+ oscillations in exocrine cells evoked by agonists and inositol trisphosphate. Cell. 1993 Aug 27;74(4):661–668. doi: 10.1016/0092-8674(93)90513-p. [DOI] [PubMed] [Google Scholar]
  42. Thorn P., Moreton R., Berridge M. Multiple, coordinated Ca2+ -release events underlie the inositol trisphosphate-induced local Ca2+ spikes in mouse pancreatic acinar cells. EMBO J. 1996 Mar 1;15(5):999–1003. [PMC free article] [PubMed] [Google Scholar]
  43. Thorn P., Petersen O. H. Activation of nonselective cation channels by physiological cholecystokinin concentrations in mouse pancreatic acinar cells. J Gen Physiol. 1992 Jul;100(1):11–25. doi: 10.1085/jgp.100.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Toescu E. C., Lawrie A. M., Petersen O. H., Gallacher D. V. Spatial and temporal distribution of agonist-evoked cytoplasmic Ca2+ signals in exocrine acinar cells analysed by digital image microscopy. EMBO J. 1992 Apr;11(4):1623–1629. doi: 10.1002/j.1460-2075.1992.tb05208.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Toescu E. C., O'Neill S. C., Petersen O. H., Eisner D. A. Caffeine inhibits the agonist-evoked cytosolic Ca2+ signal in mouse pancreatic acinar cells by blocking inositol trisphosphate production. J Biol Chem. 1992 Nov 25;267(33):23467–23470. [PubMed] [Google Scholar]
  46. Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
  47. Wakui M., Potter B. V., Petersen O. H. Pulsatile intracellular calcium release does not depend on fluctuations in inositol trisphosphate concentration. Nature. 1989 May 25;339(6222):317–320. doi: 10.1038/339317a0. [DOI] [PubMed] [Google Scholar]
  48. Wileman T., Kane L. P., Carson G. R., Terhorst C. Depletion of cellular calcium accelerates protein degradation in the endoplasmic reticulum. J Biol Chem. 1991 Mar 5;266(7):4500–4507. [PubMed] [Google Scholar]
  49. Yule D. I., Lawrie A. M., Gallacher D. V. Acetylcholine and cholecystokinin induce different patterns of oscillating calcium signals in pancreatic acinar cells. Cell Calcium. 1991 Feb-Mar;12(2-3):145–151. doi: 10.1016/0143-4160(91)90016-8. [DOI] [PubMed] [Google Scholar]

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