Chapter title |
Imaging Synaptic Vesicle Exocytosis-Endocytosis with pH-Sensitive Fluorescent Proteins.
|
---|---|
Chapter number | 11 |
Book title |
High-Resolution Imaging of Cellular Proteins
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-6352-2_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6350-8, 978-1-4939-6352-2
|
Authors |
Olusoji A. T. Afuwape, Ege T. Kavalali Ph.D., Ege T. Kavalali |
Editors |
Steven D. Schwartzbach, Omar Skalli, Thomas Schikorski |
Abstract |
The introduction of pHluorin, a pH-sensitive GFP, by Miesenbock and colleagues provided a versatile tool to studies of vesicle trafficking, in particular synaptic vesicle exocytosis and endocytosis. By tagging pHluorin to the luminal region of the synaptic vesicular protein synaptobrevin (also called VAMP, vesicle-associated membrane protein) or other synaptic vesicle-specific proteins such as the vesicular glutamate transporter-1, we are able to directly track synaptic vesicle endocytosis in response to stimuli in a molecularly specific manner. Here, we describe the process of imaging synaptic vesicle endocytosis in response to extracellular stimulation in dissociated neuronal cultures of hippocampal neurons obtained from rats-also applicable to mice-using pHluorin-tagged vesicular glutamate transporter-1 as a reporter. |
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