Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1998 Sep;75(3):1237–1243. doi: 10.1016/S0006-3495(98)74043-3

Evaluation of the electrostatic field strength at the site of exocytosis in adrenal chromaffin cells.

K Rosenheck 1
PMCID: PMC1299799  PMID: 9726926

Abstract

Exocytosis in secretory cells consists of release from intracellular storage granules directly into the extracellular space via fusion of the granule membrane with the plasma membrane of the cell. It is considered here as comprising two distinct processes. One is the close apposition of granule and plasma membranes. The other arises from interactions between the two membranes during the process of apposition, leading to the formation of a fusion pore. In the following it is shown for the case of the adrenal medullary chromaffin cell that the fusion pore can be ascribed to electroporation of the granule membrane, triggered by the strong electric field existing at the site of exocytosis. Based on an electric surface charge model of the cytoplasmic side of the plasma membrane, resulting from the negatively charged phosphatidylserine groups, it is found that the electrostatic field strength at the site of exocytosis reaches values on the order of 10(8) V/m at small intermembrane distances of 3 nm and lower. The field strength increases with the size of the disc-shaped plasma membrane region generating the electric field, reaching an approximate limit for a radius of 10 nm, at a surface charge density of 5.4 x 10(-2) C/m2. According to previous experimental evaluations of threshold field strength, this field is sufficiently strong to cause membrane electroporation. This step is a precondition for the subsequent membrane fusion during the ongoing process of apposition, leading to secretion.

Full Text

The Full Text of this article is available as a PDF (77.3 KB).

Selected References

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

  1. Allan D., Kallen K. J. Transport of lipids to the plasma membrane in animal cells. Prog Lipid Res. 1993;32(2):195–219. doi: 10.1016/0163-7827(93)90015-o. [DOI] [PubMed] [Google Scholar]
  2. Alvarez de Toledo G., Fernández-Chacón R., Fernández J. M. Release of secretory products during transient vesicle fusion. Nature. 1993 Jun 10;363(6429):554–558. doi: 10.1038/363554a0. [DOI] [PubMed] [Google Scholar]
  3. Augustine G. J., Burns M. E., DeBello W. M., Pettit D. L., Schweizer F. E. Exocytosis: proteins and perturbations. Annu Rev Pharmacol Toxicol. 1996;36:659–701. doi: 10.1146/annurev.pa.36.040196.003303. [DOI] [PubMed] [Google Scholar]
  4. Azila N., Hawthorne J. N. Subcellular localization of phospholipid changes in response to muscarinic stimulation of perfused bovine adrenal medulla. Biochem J. 1982 Apr 15;204(1):291–299. doi: 10.1042/bj2040291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Cevc G. Membrane electrostatics. Biochim Biophys Acta. 1990 Oct 8;1031(3):311–382. doi: 10.1016/0304-4157(90)90015-5. [DOI] [PubMed] [Google Scholar]
  7. Chandler D. E., Heuser J. E. Arrest of membrane fusion events in mast cells by quick-freezing. J Cell Biol. 1980 Aug;86(2):666–674. doi: 10.1083/jcb.86.2.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chernomordik L., Kozlov M. M., Zimmerberg J. Lipids in biological membrane fusion. J Membr Biol. 1995 Jul;146(1):1–14. doi: 10.1007/BF00232676. [DOI] [PubMed] [Google Scholar]
  9. Chow R. H., von Rüden L., Neher E. Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature. 1992 Mar 5;356(6364):60–63. doi: 10.1038/356060a0. [DOI] [PubMed] [Google Scholar]
  10. Crowley J. M. Electrical breakdown of bimolecular lipid membranes as an electromechanical instability. Biophys J. 1973 Jul;13(7):711–724. doi: 10.1016/S0006-3495(73)86017-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heinemann C., Chow R. H., Neher E., Zucker R. S. Kinetics of the secretory response in bovine chromaffin cells following flash photolysis of caged Ca2+. Biophys J. 1994 Dec;67(6):2546–2557. doi: 10.1016/S0006-3495(94)80744-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Helm C. A., Israelachvili J. N., McGuiggan P. M. Role of hydrophobic forces in bilayer adhesion and fusion. Biochemistry. 1992 Feb 18;31(6):1794–1805. doi: 10.1021/bi00121a030. [DOI] [PubMed] [Google Scholar]
  13. Hibino M., Shigemori M., Itoh H., Nagayama K., Kinosita K., Jr Membrane conductance of an electroporated cell analyzed by submicrosecond imaging of transmembrane potential. Biophys J. 1991 Jan;59(1):209–220. doi: 10.1016/S0006-3495(91)82212-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hui S. W. Effects of pulse length and strength on electroporation efficiency. Methods Mol Biol. 1995;55:29–40. doi: 10.1385/0-89603-328-7:29. [DOI] [PubMed] [Google Scholar]
  15. Jankowski J. A., Finnegan J. M., Wightman R. M. Extracellular ionic composition alters kinetics of vesicular release of catecholamines and quantal size during exocytosis at adrenal medullary cells. J Neurochem. 1994 Nov;63(5):1739–1747. doi: 10.1046/j.1471-4159.1994.63051739.x. [DOI] [PubMed] [Google Scholar]
  16. Levadny V. G., Belaya M. L., Pink D. A., Jericho M. H. Theory of electrostatic effects in soft biological interfaces using atomic force microscopy. Biophys J. 1996 Apr;70(4):1745–1752. doi: 10.1016/S0006-3495(96)79737-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindner P., Neumann E., Rosenheck K. Kinetics of permeability changes induced by electric impulses in chromaffin granules. J Membr Biol. 1977 Apr 22;32(3-4):231–254. doi: 10.1007/BF01905221. [DOI] [PubMed] [Google Scholar]
  18. Lopez A., Rols M. P., Teissie J. 31P NMR analysis of membrane phospholipid organization in viable, reversibly electropermeabilized Chinese hamster ovary cells. Biochemistry. 1988 Feb 23;27(4):1222–1228. doi: 10.1021/bi00404a023. [DOI] [PubMed] [Google Scholar]
  19. Marsh D. Water adsorption isotherms and hydration forces for lysolipids and diacyl phospholipids. Biophys J. 1989 Jun;55(6):1093–1100. doi: 10.1016/S0006-3495(89)82906-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marszalek P. E., Farrell B., Verdugo P., Fernandez J. M. Kinetics of release of serotonin from isolated secretory granules. I. Amperometric detection of serotonin from electroporated granules. Biophys J. 1997 Sep;73(3):1160–1168. doi: 10.1016/S0006-3495(97)78148-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McLaughlin S. The electrostatic properties of membranes. Annu Rev Biophys Biophys Chem. 1989;18:113–136. doi: 10.1146/annurev.bb.18.060189.000553. [DOI] [PubMed] [Google Scholar]
  22. Mir L. M., Orlowski S., Belehradek J., Jr, Paoletti C. Electrochemotherapy potentiation of antitumour effect of bleomycin by local electric pulses. Eur J Cancer. 1991;27(1):68–72. doi: 10.1016/0277-5379(91)90064-k. [DOI] [PubMed] [Google Scholar]
  23. Monck J. R., Fernandez J. M. The exocytotic fusion pore. J Cell Biol. 1992 Dec;119(6):1395–1404. doi: 10.1083/jcb.119.6.1395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nanavati C., Markin V. S., Oberhauser A. F., Fernandez J. M. The exocytotic fusion pore modeled as a lipidic pore. Biophys J. 1992 Oct;63(4):1118–1132. doi: 10.1016/S0006-3495(92)81679-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Needham D., Hochmuth R. M. Electro-mechanical permeabilization of lipid vesicles. Role of membrane tension and compressibility. Biophys J. 1989 May;55(5):1001–1009. doi: 10.1016/S0006-3495(89)82898-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Neumann E., Rosenheck K. Permeability changes induced by electric impulses in vesicular membranes. J Membr Biol. 1972 Dec 29;10(3):279–290. doi: 10.1007/BF01867861. [DOI] [PubMed] [Google Scholar]
  28. Neumann E., Schaefer-Ridder M., Wang Y., Hofschneider P. H. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1982;1(7):841–845. doi: 10.1002/j.1460-2075.1982.tb01257.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oberhauser A. F., Fernandez J. M. Patch clamp studies of single intact secretory granules. Biophys J. 1993 Nov;65(5):1844–1852. doi: 10.1016/S0006-3495(93)81246-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Peitzsch R. M., Eisenberg M., Sharp K. A., McLaughlin S. Calculations of the electrostatic potential adjacent to model phospholipid bilayers. Biophys J. 1995 Mar;68(3):729–738. doi: 10.1016/S0006-3495(95)80253-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rand R. P., Parsegian V. A. Mimicry and mechanism in phospholipid models of membrane fusion. Annu Rev Physiol. 1986;48:201–212. doi: 10.1146/annurev.ph.48.030186.001221. [DOI] [PubMed] [Google Scholar]
  32. Rosenheck K., Lindner P., Pecht I. Effect of electric fields on light-scattering and fluorescence of chromaffin granules. J Membr Biol. 1975;20(1-2):1–12. doi: 10.1007/BF01870624. [DOI] [PubMed] [Google Scholar]
  33. Rothman J. E. Mechanisms of intracellular protein transport. Nature. 1994 Nov 3;372(6501):55–63. doi: 10.1038/372055a0. [DOI] [PubMed] [Google Scholar]
  34. Rothman J. E., Söllner T. H. Throttles and dampers: controlling the engine of membrane fusion. Science. 1997 May 23;276(5316):1212–1213. doi: 10.1126/science.276.5316.1212. [DOI] [PubMed] [Google Scholar]
  35. Schmidt W., Patzak A., Lingg G., Winkler H., Plattner H. Membrane events in adrenal chromaffin cells during exocytosis: a freeze-etching analysis after rapid cryofixation. Eur J Cell Biol. 1983 Nov;32(1):31–37. [PubMed] [Google Scholar]
  36. Spruce A. E., Breckenridge L. J., Lee A. K., Almers W. Properties of the fusion pore that forms during exocytosis of a mast cell secretory vesicle. Neuron. 1990 May;4(5):643–654. doi: 10.1016/0896-6273(90)90192-i. [DOI] [PubMed] [Google Scholar]
  37. Sung W., Park P. J. Dynamics of pore growth in membranes and membrane stability. Biophys J. 1997 Oct;73(4):1797–1804. doi: 10.1016/S0006-3495(97)78210-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Söllner T., Bennett M. K., Whiteheart S. W., Scheller R. H., Rothman J. E. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell. 1993 Nov 5;75(3):409–418. doi: 10.1016/0092-8674(93)90376-2. [DOI] [PubMed] [Google Scholar]
  39. Teissie J., Knutson V. P., Tsong T. Y., Lane M. D. Electric pulse-induced fusion of 3T3 cells in monolayer culture. Science. 1982 Apr 30;216(4545):537–538. doi: 10.1126/science.7071601. [DOI] [PubMed] [Google Scholar]
  40. Teissie J., Tsong T. Y. Electric field induced transient pores in phospholipid bilayer vesicles. Biochemistry. 1981 Mar 17;20(6):1548–1554. doi: 10.1021/bi00509a022. [DOI] [PubMed] [Google Scholar]
  41. Tsong T. Y. Electric modification of membrane permeability for drug loading into living cells. Methods Enzymol. 1987;149:248–259. doi: 10.1016/0076-6879(87)49063-0. [DOI] [PubMed] [Google Scholar]
  42. Tsong T. Y. Electroporation of cell membranes. Biophys J. 1991 Aug;60(2):297–306. doi: 10.1016/S0006-3495(91)82054-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vitale M. L., Seward E. P., Trifaró J. M. Chromaffin cell cortical actin network dynamics control the size of the release-ready vesicle pool and the initial rate of exocytosis. Neuron. 1995 Feb;14(2):353–363. doi: 10.1016/0896-6273(95)90291-0. [DOI] [PubMed] [Google Scholar]
  44. Wightman R. M., Jankowski J. A., Kennedy R. T., Kawagoe K. T., Schroeder T. J., Leszczyszyn D. J., Near J. A., Diliberto E. J., Jr, Viveros O. H. Temporally resolved catecholamine spikes correspond to single vesicle release from individual chromaffin cells. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10754–10758. doi: 10.1073/pnas.88.23.10754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wilson S. P., Kirshner N. Isolation and characterization of plasma membranes from the adrenal medulla. J Neurochem. 1976 Dec;27(6):1289–1298. doi: 10.1111/j.1471-4159.1976.tb02606.x. [DOI] [PubMed] [Google Scholar]
  46. Zimmermann U. Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta. 1982 Nov 30;694(3):227–277. doi: 10.1016/0304-4157(82)90007-7. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES