Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1991 June 1; 113(5): 1057–1067.
PMCID: PMC2289017

Cortical filamentous actin disassembly and scinderin redistribution during chromaffin cell stimulation precede exocytosis, a phenomenon not exhibited by gelsolin


Immunofluorescence and cytochemical studies have demonstrated that filamentous actin is mainly localized in the cortical surface of the chromaffin cell. It has been suggested that these actin filament networks act as a barrier to the secretory granules, impeding their contact with the plasma membrane. Stimulation of chromaffin cells produces a disassembly of actin filament networks, implying the removal of the barrier. The presence of gelsolin and scinderin, two Ca(2+)- dependent actin filament severing proteins, in the cortical surface of the chromaffin cells, suggests the possibility that cell stimulation brings about activation of one or more actin filament severing proteins with the consequent disruption of actin networks. Therefore, biochemical studies and fluorescence microscopy experiments with scinderin and gelsolin antibodies and rhodamine-phalloidin, a probe for filamentous actin, were performed in cultured chromaffin cells to study the distribution of scinderin, gelsolin, and filamentous actin during cell stimulation and to correlate the possible changes with catecholamine secretion. Here we report that during nicotinic stimulation or K(+)-evoked depolarization, subcortical scinderin but not gelsolin is redistributed and that this redistribution precedes catecholamine secretion. The rearrangement of scinderin in patches is mediated by nicotinic receptors. Cell stimulation produces similar patterns of distribution of scinderin and filamentous actin. However, after the removal of the stimulus, the recovery of scinderin cortical pattern of distribution is faster than F-actin reassembly, suggesting that scinderin is bound in the cortical region of the cell to a component other than F-actin. We also demonstrate that peripheral actin filament disassembly and subplasmalemmal scinderin redistribution are calcium-dependent events. Moreover, experiments with an antibody against dopamine-beta-hydroxylase suggest that exocytosis sites are preferentially localized to areas of F-actin disassembly.

Full Text

The Full Text of this article is available as a PDF (1.8M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aunis D, Guerold B, Bader MF, Cieselski-Treska J. Immunocytochemical and biochemical demonstration of contractile proteins in chromaffin cells in culture. Neuroscience. 1980;5(12):2261–2277. [PubMed]
  • Bader MF, Georges E, Mushynski WE, Trifaró JM. Neurofilament proteins in cultured chromaffin cells. J Neurochem. 1984 Oct;43(4):1180–1193. [PubMed]
  • Bader MF, Trifaró JM, Langley OK, Thiersé D, Aunis D. Secretory cell actin-binding proteins: identification of a gelsolin-like protein in chromaffin cells. J Cell Biol. 1986 Feb;102(2):636–646. [PMC free article] [PubMed]
  • Bader MF, Sontag JM, Thiersé D, Aunis D. A reassessment of guanine nucleotide effects on catecholamine secretion from permeabilized adrenal chromaffin cells. J Biol Chem. 1989 Oct 5;264(28):16426–16434. [PubMed]
  • Baker EM, Cheek TR, Burgoyne RD. Cyclic AMP inhibits secretion from bovine adrenal chromaffin cells evoked by carbamylcholine but not by high K+. Biochim Biophys Acta. 1985 Sep 30;846(3):388–393. [PubMed]
  • Bernstein BW, Bamburg JR. Reorganization of actin in depolarized synaptosomes. J Neurosci. 1985 Oct;5(10):2565–2569. [PubMed]
  • Bittner MA, Holz RW. Phorbol esters enhance exocytosis from chromaffin cells by two mechanisms. J Neurochem. 1990 Jan;54(1):205–210. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Burgoyne RD, Cheek TR. Reorganisation of peripheral actin filaments as a prelude to exocytosis. Biosci Rep. 1987 Apr;7(4):281–288. [PubMed]
  • Burgoyne RD, Cheek TR, Norman KM. Identification of a secretory granule-binding protein as caldesmon. Nature. 1986 Jan 2;319(6048):68–70. [PubMed]
  • Burgoyne RD, Morgan A, O'Sullivan AJ. The control of cytoskeletal actin and exocytosis in intact and permeabilized adrenal chromaffin cells: role of calcium and protein kinase C. Cell Signal. 1989;1(4):323–334. [PubMed]
  • Chaponnier C, Janmey PA, Yin HL. The actin filament-severing domain of plasma gelsolin. J Cell Biol. 1986 Oct;103(4):1473–1481. [PMC free article] [PubMed]
  • Cheek TR, Burgoyne RD. Nicotine-evoked disassembly of cortical actin filaments in adrenal chromaffin cells. FEBS Lett. 1986 Oct 20;207(1):110–114. [PubMed]
  • Cheek TR, Burgoyne RD. Cyclic AMP inhibits both nicotine-induced actin disassembly and catecholamine secretion from bovine adrenal chromaffin cells. J Biol Chem. 1987 Aug 25;262(24):11663–11666. [PubMed]
  • Cheek TR, Jackson TR, O'Sullivan AJ, Moreton RB, Berridge MJ, Burgoyne RD. Simultaneous measurements of cytosolic calcium and secretion in single bovine adrenal chromaffin cells by fluorescent imaging of fura-2 in cocultured cells. J Cell Biol. 1989 Sep;109(3):1219–1227. [PMC free article] [PubMed]
  • Côté A, Doucet JP, Trifaró JM. Phosphorylation and dephosphorylation of chromaffin cell proteins in response to stimulation. Neuroscience. 1986 Oct;19(2):629–645. [PubMed]
  • Doucet JP, Trifaró JM. A discontinuous and highly porous sodium dodecyl sulfate-polyacrylamide slab gel system of high resolution. Anal Biochem. 1988 Feb 1;168(2):265–271. [PubMed]
  • Douglas WW. Stimulus-secretion coupling: the concept and clues from chromaffin and other cells. Br J Pharmacol. 1968 Nov;34(3):451–474. [PMC free article] [PubMed]
  • DOUGLAS WW, RUBIN RP. The role of calcium in the secretory response of the adrenal medulla to acetylcholine. J Physiol. 1961 Nov;159:40–57. [PubMed]
  • Faulstich H, Zobeley S, Rinnerthaler G, Small JV. Fluorescent phallotoxins as probes for filamentous actin. J Muscle Res Cell Motil. 1988 Oct;9(5):370–383. [PubMed]
  • Fisher SK, Holz RW, Agranoff BW. Muscarinic receptors in chromaffin cell cultures mediate enhanced phospholipid labeling but not catecholamine secretion. J Neurochem. 1981 Aug;37(2):491–497. [PubMed]
  • Hughes AR, Putney JW., Jr Inositol phosphate formation and its relationship to calcium signaling. Environ Health Perspect. 1990 Mar;84:141–147. [PMC free article] [PubMed]
  • Joh TH, Hwang O. Dopamine beta-hydroxylase: biochemistry and molecular biology. Ann N Y Acad Sci. 1987;493:342–350. [PubMed]
  • Kao LS, Schneider AS. Muscarinic receptors on bovine chromaffin cells mediate a rise in cytosolic calcium that is independent of extracellular calcium. J Biol Chem. 1985 Feb 25;260(4):2019–2022. [PubMed]
  • Kenigsberg RL, Trifaró JM. Presence of a high affinity uptake system for catecholamines in cultured bovine adrenal chromaffin cells. Neuroscience. 1980;5(9):1547–1556. [PubMed]
  • Kenigsberg RL, Trifaró JM. Microinjection of calmodulin antibodies into cultured chromaffin cells blocks catecholamine release in response to stimulation. Neuroscience. 1985 Jan;14(1):335–347. [PubMed]
  • Kim KT, Westhead EW. Cellular responses to Ca2+ from extracellular and intracellular sources are different as shown by simultaneous measurements of cytosolic Ca2+ and secretion from bovine chromaffin cells. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9881–9885. [PubMed]
  • Koffer A, Tatham PE, Gomperts BD. Changes in the state of actin during the exocytotic reaction of permeabilized rat mast cells. J Cell Biol. 1990 Sep;111(3):919–927. [PMC free article] [PubMed]
  • Lee RW, Trifaró JM. Characterization of anti-actin antibodies and their use in immunocytochemical studies on the localization of actin in adrenal chromaffin cells in culture. Neuroscience. 1981;6(10):2087–2108. [PubMed]
  • Maekawa S, Sakai H. Inhibition of actin regulatory activity of the 74-kDa protein from bovine adrenal medulla (adseverin) by some phospholipids. J Biol Chem. 1990 Jul 5;265(19):10940–10942. [PubMed]
  • Maekawa S, Toriyama M, Hisanaga S, Yonezawa N, Endo S, Hirokawa N, Sakai H. Purification and characterization of a Ca2+-dependent actin filament severing protein from bovine adrenal medulla. J Biol Chem. 1989 May 5;264(13):7458–7465. [PubMed]
  • Matter K, Dreyer F, Aktories K. Actin involvement in exocytosis from PC12 cells: studies on the influence of botulinum C2 toxin on stimulated noradrenaline release. J Neurochem. 1989 Feb;52(2):370–376. [PubMed]
  • O'Sullivan AJ, Cheek TR, Moreton RB, Berridge MJ, Burgoyne RD. Localization and heterogeneity of agonist-induced changes in cytosolic calcium concentration in single bovine adrenal chromaffin cells from video imaging of fura-2. EMBO J. 1989 Feb;8(2):401–411. [PubMed]
  • Perrin D, Aunis D. Reorganization of alpha-fodrin induced by stimulation in secretory cells. Nature. 1985 Jun 13;315(6020):589–592. [PubMed]
  • Phillips JH, Burridge K, Wilson SP, Kirshner N. Visualization of the exocytosis/endocytosis secretory cycle in cultured adrenal chromaffin cells. J Cell Biol. 1983 Dec;97(6):1906–1917. [PMC free article] [PubMed]
  • Rodriguez Del Castillo A, Lemaire S, Tchakarov L, Jeyapragasan M, Doucet JP, Vitale ML, Trifaró JM. Chromaffin cell scinderin, a novel calcium-dependent actin filament-severing protein. EMBO J. 1990 Jan;9(1):43–52. [PubMed]
  • Stossel TP, Chaponnier C, Ezzell RM, Hartwig JH, Janmey PA, Kwiatkowski DJ, Lind SE, Smith DB, Southwick FS, Yin HL, et al. Nonmuscle actin-binding proteins. Annu Rev Cell Biol. 1985;1:353–402. [PubMed]
  • Tchakarov L, Vitale ML, Jeyapragasan M, Rodriguez Del Castillo A, Trifaró JM. Expression of scinderin, an actin filament-severing protein, in different tissues. FEBS Lett. 1990 Jul 30;268(1):209–212. [PubMed]
  • TerBush DR, Bittner MA, Holz RW. Ca2+ influx causes rapid translocation of protein kinase C to membranes. Studies of the effects of secretagogues in adrenal chromaffin cells. J Biol Chem. 1988 Dec 15;263(35):18873–18879. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Trifaró JM. Common mechanisms of hormone secretion. Annu Rev Pharmacol Toxicol. 1977;17:27–47. [PubMed]
  • Trifaró JM, Lee RW. Morphological characteristics and stimulus-secretion coupling in bovine adcrenal chromaffin cell cultures. Neuroscience. 1980;5(9):1533–1546. [PubMed]
  • Trifaró JM, Bourne GW. Differential effects of concanavalin A on acetylcholine and potassium-evoked release of catecholamines from cultured chromaffin cells. Neuroscience. 1981;6(9):1823–1833. [PubMed]
  • Trifaró JM, Duerr AC, Pinto JE. Membranes of the adrenal medulla: a comparison between the membranes of the Golgi apparatus and chromaffin granules. Mol Pharmacol. 1976 Jul;12(4):536–545. [PubMed]
  • Trifaró JM, Kenigsberg RL, Côté A, Lee RW, Hikita T. Adrenal paraneurone contractile proteins and stimulus-secretion coupling. Can J Physiol Pharmacol. 1984 Apr;62(4):493–501. [PubMed]
  • Trifaró JM, Bader MF, Doucet JP. Chromaffin cell cytoskeleton: its possible role in secretion. Can J Biochem Cell Biol. 1985 Jun;63(6):661–679. [PubMed]
  • Wilson SP, Kirshner N. The acetylcholine receptor of the adrenal medulla. J Neurochem. 1977 Apr;28(4):687–695. [PubMed]
  • Yin HL, Stossel TP. Control of cytoplasmic actin gel-sol transformation by gelsolin, a calcium-dependent regulatory protein. Nature. 1979 Oct 18;281(5732):583–586. [PubMed]
  • Yin HL, Albrecht JH, Fattoum A. Identification of gelsolin, a Ca2+-dependent regulatory protein of actin gel-sol transformation, and its intracellular distribution in a variety of cells and tissues. J Cell Biol. 1981 Dec;91(3 Pt 1):901–906. [PMC free article] [PubMed]
  • Yin HL, Iida K, Janmey PA. Identification of a polyphosphoinositide-modulated domain in gelsolin which binds to the sides of actin filaments. J Cell Biol. 1988 Mar;106(3):805–812. [PMC free article] [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press