title: A nibbling mechanism for clathrin-mediated retrieval of secretory granule membrane after exocytosis.
abstract: Clathrin-mediated endocytosis is the major pathway for recycling of granule membrane components after strong stimulation and high exocytotic rates. It resembles "classical" receptor-mediated endocytosis but has a trigger that is unique to secretion, the sudden appearance of the secretory granule membrane in the plasma membrane. The spatial localization, the relationship to individual fusion events, the nature of the cargo, and the timing and nature of the nucleation events are unknown. Furthermore, a size mismatch between chromaffin granules (∼300-nm diameter) and typical clathrin-coated vesicles (∼90 nm) makes it unlikely that clathrin-mediated endocytosis internalizes as a unit the entire fused granule membrane. We have used a combination of total internal reflection fluorescence microscopy of transiently expressed proteins and time-resolved quantitative confocal imaging of endogenous proteins along with a fluid-phase marker to address these issues. We demonstrate that the fused granule membrane remains a distinct entity and serves as a nucleation site for clathrin- and dynamin-mediated endocytosis that internalizes granule membrane components in small increments.
authors: Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109-5632, USA. mbittner@umich.edu
authors: Mary A
authors: MA
authors: Bittner
authors: Rachel L
authors: RL
authors: Aikman
authors: Ronald W
authors: RW
authors: Holz
mesh_terms: D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D019439:Chromaffin Cells / Q000166:cytology; D002837:Chromaffin Granules / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D004705:Endocytosis; D005089:Exocytosis / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D008561:Membrane Fusion; D018613:Microscopy, Confocal / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008954:Models, Biological; D055099:Neuroendocrine Cells / Q000166:cytology; D022142:Secretory Vesicles / Q000378:metabolism*; D014162:Transfection
chemical_list: D002966:Clathrin; D049452:Green Fluorescent Proteins; D004299:Dopamine beta-Hydroxylase
references: J Cell Biol. 1973 Feb;56(2):540-58
references: 4345556
references: J Biol Chem. 2011 Oct 14;286(41):35966-35976
references: 21832053
references: Nat Cell Biol. 2002 Sep;4(9):691-8
references: 12198492
references: J Cell Biol. 1976 Aug;70(2 pt 1):294-303
references: 181385
references: Traffic. 2011 Dec;12(12):1868-78
references: 21883765
references: Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20576-81
references: 18077369
references: Philos Trans R Soc Lond B Biol Sci. 1971 Jun 17;261(839):291-2
references: 4399621
references: J Neurosci. 2005 Aug 10;25(32):7324-32
references: 16093382
references: Mol Biol Cell. 2004 Oct;15(10):4658-68
references: 15282339
references: Biochem J. 1970 Jun;118(2):303-10
references: 4320820
references: Nature. 2003 Mar 6;422(6927):37-44
references: 12621426
references: J Biol Chem. 2000 Jun 9;275(23):17878-85
references: 10747966
references: Mol Biol Cell. 2011 Jun 1;22(11):1907-18
references: 21460182
references: Nature. 1968 Jan 27;217(5126):384-8
references: 4867231
references: Traffic. 2011 Jan;12(1):72-88
references: 20880191
references: J Cell Biol. 2010 Feb 8;188(3):415-28
references: 20142424
references: J Cell Biol. 1976 Jul;70(1):59-74
references: 932102
references: Naunyn Schmiedebergs Arch Pharmacol. 1972;273(1):43-61
references: 4260804
references: Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8328-32
references: 7667289
references: Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):6358-63
references: 11959911
references: J Cell Biol. 1983 Dec;97(6):1906-17
references: 6643581
references: J Biol Chem. 1993 May 25;268(15):10983-9
references: 8496162
references: J Cell Biol. 1986 Feb;102(2):510-5
references: 3080437
references: Cell. 1994 Sep 9;78(5):751-60
references: 8087843
references: PLoS Biol. 2011 Mar;9(3):e1000604
references: 21445324
references: J Biol Chem. 1998 Jul 10;273(28):17351-60
references: 9651318
references: Science. 2007 Apr 27;316(5824):570-4
references: 17463283
references: Neuroscience. 1980;5(11):1803-23
references: 7432623
references: Proc Natl Acad Sci U S A. 1977 Nov;74(11):5073-7
references: 270741
references: Curr Opin Neurobiol. 1996 Jun;6(3):350-7
references: 8794084
references: Science. 1999 Aug 20;285(5431):1268-71
references: 10455054
references: Nature. 1984 Mar 8-14;308(5955):208-10
references: 6422310
references: Mol Biol Cell. 2008 Dec;19(12):5347-59
references: 18923138
references: J Cell Biol. 1984 May;98(5):1817-24
references: 6373784
references: Neuron. 2006 Sep 21;51(6):773-86
references: 16982422
references: Exp Cell Res. 1989 Jun;182(2):445-60
references: 2721590
references: Cell Tissue Res. 1984;235(2):365-70
references: 6367999
references: Neuroscience. 1997 Oct;80(3):847-60
references: 9276499
references: Biochim Biophys Acta. 1971 Sep 1;239(3):517-9
references: 4329717
references: Brain Res. 1972 Feb 11;37(1):141-5
references: 5060888