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. 1988 Dec;406:299–313. doi: 10.1113/jphysiol.1988.sp017381

Agonist-induced changes in cell membrane capacitance and conductance in dialysed pancreatic acinar cells of rats.

Y Maruyama 1
PMCID: PMC1191100  PMID: 2474070

Abstract

1. Single acinar cells enzymatically isolated from the rat pancreas were subjected to tight-seal whole-cell recordings. Changes in cell membrane capacitance and conductance were simultaneously recorded using a phase-sensitive detection method. 2. Acetylcholine (ACh, 0.05-0.5 microM) and cholecystokinin octapeptide (CCK-8, 10-50 pM) concomitantly induced transient increases in cell membrane current, capacitance and conductance only when cytosolic Ca2+ was weakly chelated by EGTA (70 microM). These responses were prolonged when the cells were dialysed with a solution containing GTP gamma S (a stable analogue of GTP, 50-100 microM), whereas they were inhibited by dialysing with that containing GDP beta S (a stable analogue of GDP). These results suggest that a type of guanine-nucleotide-binding protein (G-protein) could be involved in ACh- or CCK-receptor signalling. 3. The ACh- or CCK-induced responses (with or without GTP gamma S in the cytosol) were all abolished when a high dose of EGTA (1-2 mM) was injected into the acinar cells. In addition, A23187, a calcium ionophore, induced sustained responses when the cytosolic Ca2+ was weakly buffered by 70 microM-EGTA. These results suggest that the secretagogues regulate the changes in cell membrane capacitance and conductance via an increase and decrease of cytosolic Ca2+ concentration. 4. Oscillatory changes in cell membrane conductance and capacitance were consistently observed even without applying secretagogues when the cells were dialysed with a solution containing GTP gamma S (50-100 microM) and cytosolic free Ca2+ ions weakly buffered at about 10(-6) M with a low dose of EGTA and CaCl2. 5. The peak amplitude of changes in cell membrane capacitance induced by ACh or CCK-8, with or without GTP gamma S in the cytosol, varied between 200 and 1000 fF, thereby suggesting that 20-100 zymogen granules can fuse with the luminal cell membrane in response to these agonists in rat exocrine pancreatic acinar cells.

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Selected References

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  1. Almers W., Neher E. Gradual and stepwise changes in the membrane capacitance of rat peritoneal mast cells. J Physiol. 1987 May;386:205–217. doi: 10.1113/jphysiol.1987.sp016530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amsterdam A., Jamieson J. D. Studies on dispersed pancreatic exocrine cells. I. Dissociation technique and morphologic characteristics of separated cells. J Cell Biol. 1974 Dec;63(3):1037–1056. doi: 10.1083/jcb.63.3.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amsterdam A., Jamieson J. D. Studies on dispersed pancreatic exocrine cells. II. Functional characteristics of separated cells. J Cell Biol. 1974 Dec;63(3):1057–1073. doi: 10.1083/jcb.63.3.1057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  5. Bolender R. P. Stereological analysis of the guinea pig pancreas. I. Analytical model and quantitative description of nonstimulated pancreatic exocrine cells. J Cell Biol. 1974 May;61(2):269–287. doi: 10.1083/jcb.61.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Breckenridge L. J., Almers W. Currents through the fusion pore that forms during exocytosis of a secretory vesicle. 1987 Aug 27-Sep 2Nature. 328(6133):814–817. doi: 10.1038/328814a0. [DOI] [PubMed] [Google Scholar]
  7. Clapham D. E., Neher E. Trifluoperazine reduces inward ionic currents and secretion by separate mechanisms in bovine chromaffin cells. J Physiol. 1984 Aug;353:541–564. doi: 10.1113/jphysiol.1984.sp015350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dockray G. J. The action of scretin, cholecystokinin-pancreozymin and caerulein on pancreatic secretion in the rat. J Physiol. 1972 Sep;225(3):679–692. doi: 10.1113/jphysiol.1972.sp009963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans M. G., Marty A. Potentiation of muscarinic and alpha-adrenergic responses by an analogue of guanosine 5'-triphosphate. Proc Natl Acad Sci U S A. 1986 Jun;83(11):4099–4103. doi: 10.1073/pnas.83.11.4099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fernandez J. M., Neher E., Gomperts B. D. Capacitance measurements reveal stepwise fusion events in degranulating mast cells. 1984 Nov 29-Dec 5Nature. 312(5993):453–455. doi: 10.1038/312453a0. [DOI] [PubMed] [Google Scholar]
  11. Iwatsuki N., Petersen O. H. Pancreatic acinar cells: the acetylcholine equilibrium potential and its ionic dependency. J Physiol. 1977 Aug;269(3):735–751. doi: 10.1113/jphysiol.1977.sp011926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jauch P., Petersen O. H., Läuger P. Electrogenic properties of the sodium-alanine cotransporter in pancreatic acinar cells: I. Tight-seal whole-cell recordings. J Membr Biol. 1986;94(2):99–115. doi: 10.1007/BF01871191. [DOI] [PubMed] [Google Scholar]
  13. Knight D. E., Baker P. F. Calcium-dependence of catecholamine release from bovine adrenal medullary cells after exposure to intense electric fields. J Membr Biol. 1982;68(2):107–140. doi: 10.1007/BF01872259. [DOI] [PubMed] [Google Scholar]
  14. Maruyama Y. A patch-clamp study of mammalian platelets and their voltage-gated potassium current. J Physiol. 1987 Oct;391:467–485. doi: 10.1113/jphysiol.1987.sp016750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maruyama Y. Ca2+-induced excess capacitance fluctuation studied by phase-sensitive detection method in exocrine pancreatic acinar cells. Pflugers Arch. 1986 Nov;407(5):561–563. doi: 10.1007/BF00657517. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Maruyama Y., Petersen O. H. Single calcium-dependent cation channels in mouse pancreatic acinar cells. J Membr Biol. 1984;81(1):83–87. doi: 10.1007/BF01868812. [DOI] [PubMed] [Google Scholar]
  18. Maruyama Y., Peterson O. H. Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini. Nature. 1982 Sep 9;299(5879):159–161. doi: 10.1038/299159a0. [DOI] [PubMed] [Google Scholar]
  19. Merritt J. E., Taylor C. W., Rubin R. P., Putney J. W., Jr Evidence suggesting that a novel guanine nucleotide regulatory protein couples receptors to phospholipase C in exocrine pancreas. Biochem J. 1986 Jun 1;236(2):337–343. doi: 10.1042/bj2360337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milutinović S., Argent B. E., Schulz U., Sachs G. Studies on isolated subcellular components of cat pancreas. II. A Ca++-dependent interaction between membranes and zymogen granules of cat pancreas. J Membr Biol. 1977 Sep 14;36(2-3):281–295. doi: 10.1007/BF01868155. [DOI] [PubMed] [Google Scholar]
  21. Neher E., Marty A. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6712–6716. doi: 10.1073/pnas.79.21.6712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Norman P. S., Mann G. E. Ionic dependence of amino-acid transport in the exocrine pancreatic epithelium: calcium dependence of insulin action. J Membr Biol. 1987;96(2):153–163. doi: 10.1007/BF01869241. [DOI] [PubMed] [Google Scholar]
  23. Palade G. Intracellular aspects of the process of protein synthesis. Science. 1975 Aug 1;189(4200):347–358. doi: 10.1126/science.1096303. [DOI] [PubMed] [Google Scholar]
  24. Petersen O. H., Maruyama Y., Graf J., Laugier R., Nishiyama A., Pearson G. T. Ionic currents across pancreatic acinar cell membranes and their role in fluid secretion. Philos Trans R Soc Lond B Biol Sci. 1981 Dec 18;296(1080):151–166. doi: 10.1098/rstb.1981.0179. [DOI] [PubMed] [Google Scholar]
  25. Randriamampita C., Chanson M., Trautmann A. Calcium and secretagogues-induced conductances in rat exocrine pancreas. Pflugers Arch. 1988 Jan;411(1):53–57. doi: 10.1007/BF00581646. [DOI] [PubMed] [Google Scholar]
  26. Streb H., Bayerdörffer E., Haase W., Irvine R. F., Schulz I. Effect of inositol-1,4,5-trisphosphate on isolated subcellular fractions of rat pancreas. J Membr Biol. 1984;81(3):241–253. doi: 10.1007/BF01868717. [DOI] [PubMed] [Google Scholar]
  27. Streb H., Irvine R. F., Berridge M. J., Schulz I. Release of Ca2+ from a nonmitochondrial intracellular store in pancreatic acinar cells by inositol-1,4,5-trisphosphate. Nature. 1983 Nov 3;306(5938):67–69. doi: 10.1038/306067a0. [DOI] [PubMed] [Google Scholar]
  28. Woods N. M., Cuthbertson K. S., Cobbold P. H. Repetitive transient rises in cytoplasmic free calcium in hormone-stimulated hepatocytes. Nature. 1986 Feb 13;319(6054):600–602. doi: 10.1038/319600a0. [DOI] [PubMed] [Google Scholar]

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