title: Resolving vesicle fusion from lysis to monitor calcium-triggered lysosomal exocytosis in astrocytes.
abstract: Optical imaging of individual vesicle exocytosis is providing new insights into the mechanism and regulation of secretion by cells. To study calcium-triggered secretion from astrocytes, we used acridine orange (AO) to label vesicles. Although AO is often used for imaging exocytosis, we found that imaging vesicles labeled with AO can result in their photolysis. Here, we define experimental and analytical approaches that permit us to distinguish unambiguously between fusion, leakage, and lysis of individual vesicles. We have used this approach to demonstrate that lysosomes undergo calcium-triggered exocytosis in astrocytes.
authors: The Rockefeller University, 1230 York Avenue, Box 304, New York, NY 10065, USA. jaiswaj@rockefeller.edu
authors: Jyoti K
authors: JK
authors: Jaiswal
authors: Marina
authors: M
authors: Fix
authors: Takahiro
authors: T
authors: Takano
authors: Maiken
authors: M
authors: Nedergaard
authors: Sanford M
authors: SM
authors: Simon
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology*; D002118:Calcium / Q000502:physiology*; D005089:Exocytosis; D008247:Lysosomes / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats
chemical_list: D002118:Calcium
references: Nat Cell Biol. 2003 Feb;5(2):126-36
references: 12545172
references: Nat Rev Neurosci. 2006 Mar;7(3):194-206
references: 16495941
references: Curr Biol. 2003 Apr 1;13(7):563-7
references: 12676086
references: Nat Biotechnol. 2004 Apr;22(4):445-9
references: 14990965
references: Nat Rev Mol Cell Biol. 2001 Apr;2(4):268-75
references: 11283724
references: PLoS Biol. 2004 Aug;2(8):E233
references: 15226824
references: J Cell Biol. 2000 Apr 3;149(1):23-32
references: 10747084
references: Drug Discov Today. 1999 Jan;4(1):32-38
references: 10234148
references: Cell. 2003 Feb 21;112(4):519-33
references: 12600315
references: J Cell Biol. 1996 Jun;133(6):1177-91
references: 8682857
references: J Exp Med. 1998 May 18;187(10):1583-98
references: 9584137
references: J Neurochem. 1999 Feb;72(2):625-33
references: 9930734
references: Nat Neurosci. 2004 Jun;7(6):613-20
references: 15156145
references: Glia. 2006 Nov 15;54(7):700-715
references: 17006898
references: J Cell Biol. 2000 Jan 24;148(2):317-24
references: 10648564
references: Cell. 2001 Jul 27;106(2):157-69
references: 11511344
references: Trends Neurosci. 2003 Oct;26(10):523-30
references: 14522144
references: Nat Chem Biol. 2007 Feb;3(2):92-8
references: 17235347
references: Neuron. 1997 Jun;18(6):857-63
references: 9208853
references: J Cell Biol. 2001 Apr 2;153(1):177-90
references: 11285284
references: Biophys J. 2002 Jul;83(1):172-83
references: 12080110
references: Traffic. 2001 Nov;2(11):764-74
references: 11733042
references: Photochem Photobiol. 1990 Jan;51(1):67-76
references: 2304980
references: Trends Neurosci. 2003 Oct;26(10):520-2
references: 14522143
references: J Photochem Photobiol B. 2007 Jan 3;86(1):1-8
references: 16982198
references: Proc Natl Acad Sci U S A. 2004 May 11;101(19):7311-6
references: 15123811
references: EMBO J. 1996 Aug 1;15(15):3787-91
references: 8670883
references: Science. 2004 Jul 2;305(5680):55-9
references: 15232098
references: Mol Biol Cell. 2001 Nov;12(11):3489-501
references: 11694583
references: Nature. 1997 Jul 31;388(6641):474-8
references: 9242406
references: Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):2070-5
references: 12538853
references: Biophys J. 2007 Aug 1;93(3):969-80
references: 17416619
references: J Membr Biol. 1996 Jul;152(2):159-67
references: 9139126
references: Virchows Arch B Cell Pathol Incl Mol Pathol. 1993;64(6):401-6
references: 8148962
references: Proc Natl Acad Sci U S A. 2005 Nov 8;102(45):16466-71
references: 16254051
references: Annu Rev Cell Dev Biol. 2003;19:697-731
references: 14570587
references: J Cell Biol. 2002 Nov 25;159(4):625-35
references: 12438417
references: Curr Biol. 2000 Oct 19;10(20):1307-10
references: 11069115