Resolving vesicle fusion from lysis to monitor calcium-triggered lysosomal exocytosis in astrocytes

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 im...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2007-08, Vol.104 (35), p.14151-14156
Main Authors: Jaiswal, Jyoti K, Fix, Marina, Takano, Takahiro, Nedergaard, Maiken, Simon, Sanford M
Format: Article
Language:English
Subjects:
Acids
Animals
Astrocytes
Astrocytes - cytology
Astrocytes - physiology
Biological Sciences
Calcium
Calcium - physiology
Cell membranes
Cells
Exocytosis
Fluorescence
Imaging
Lasers
Lysosomes
Lysosomes - physiology
Lysosomes - ultrastructure
Membranes
Microscopy
Photolysis
Pollutant emissions
Proteins
Rats
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0704935104