Pflügers Archiv

, Volume 427, Issue 3–4, pp 233–243

Induction of Ca2+ oscillations by selective, U73122-mediated, depletion of inositol-trisphosphate-sensitive Ca2+ stores in rabbit pancreatic acinar cells

  • P. H. G. M. Willems
  • F. H. M. M. Van de Put
  • R. Engbersen
  • R. R. Bosch
  • H. J. M. Van Hoof
  • J. J. H. H. M. de Pont
Molecular and Cellular Physiology

DOI: 10.1007/BF00374529

Cite this article as:
Willems, P.H.G.M., Van de Put, F.H.M.M., Engbersen, R. et al. Pflügers Arch (1994) 427: 233. doi:10.1007/BF00374529
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Abstract

The effect of the putative inhibitor of phospholipase C activity, U73122, on the Ca2+ sequestering and releasing properties of internal Ca2+ stores was studied in both permeabilized and intact rabbit pancreatic acinar cells. U73122 dose dependently inhibited ATP-dependent Ca2+ uptake in the inositol (1,4,5)-trisphosphate-[Ins(1,4,5)P3]-sensitive, but not the Ins(1,4,5)P3-insensitive, Ca2+ store in acinar cells permeabilized by saponin treatment. In a suspension of intact acinar cells, loaded with the fluorescent Ca2+ indicator, Fura-2, U73122 alone evoked a transient increase in average free cytosolic Ca2+ concentration ([Ca2+]i,av), which was largely independent of external Ca2+. Addition of U73122 to cell suspensions prestimulated with either cholecystokinin octapeptide or JMV-180 revealed an inverse relationship in size between the U73122- and the agonistevoked [Ca2+]i,av transient. Moreover, thapsigargin-induced inhibition of intracellular Ca2+-ATPase activity resulted in a [Ca2+]i,av transient, the size of which was not different following maximal prestimulation with either U73122 or agonist. These observations suggest that U73122 selectively affects the Ins(1,4,5)P3- casu quo agonist-sensitive internal Ca2+ store, whereas thapsigargin affects both the Ins(1,4,5)P3-sensitive and -insensitive Ca2+ store. Digital-imaging microscopy of Fura-2-loaded acinar cells demonstrated that U73122, in contrast to thapsigargin, evoked sustained oscillatory changes in [Ca2+]i. The U73122-evoked oscillations were abolished in the absence of external Ca2+. The ability of U73122 to generate external Ca2+-dependent Ca2+ oscillations suggests that depletion of the agonistsensitive store leads to an increase in Ca2+ permeability of the plasma membrane and that the Ins(1,4,5)P3-insensitive Ca2+ pool is necessary for the Ca2+ oscillations.

Key words

Pancreatic acinar cell Cholecystokininoctapeptide JMV-180 U73122 Thapsigargin Ca2+ oscillations Permeabilized acinar cells 

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • P. H. G. M. Willems
    • 1
  • F. H. M. M. Van de Put
    • 1
  • R. Engbersen
    • 1
  • R. R. Bosch
    • 1
  • H. J. M. Van Hoof
    • 1
  • J. J. H. H. M. de Pont
    • 1
  1. 1.Department of BiochemistryUniversity of NijmegenHB NijmegenThe Netherlands

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