Document Detail


In vivo imaging of vesicle motion and release at the Drosophila neuromuscular junction.
MedLine Citation:
PMID:  17546002     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recently, it has become possible to directly detect changes in neuropeptide vesicle dynamics in nerve terminals in vivo and to measure the release of neuropeptides induced experimentally or evoked by normal behavior. These results were obtained with the use of transgenic fruit flies that express a neuropeptide tagged with green fluorescent protein. Here, we describe how vesicle movement and neuropeptide release can be studied in the larval Drosophila neuromuscular junction using fluorescence microscopy. Analysis methods are described for quantifying movement based on time lapse and fluorescence recovery after photobleaching data. Specific approaches that can be applied to nerve terminals include single particle tracking, correlation and Fourier analysis. Utilization of these methods led to the first detection of vesicle mobilization in nerve terminals and the discoveries of activity-dependent capture of transiting vesicles and post-tetanic potentiation of neuropeptide release. Overall, this protocol can be carried out in an hour with ready Drosophila.
Authors:
Edwin S Levitan; Frederick Lanni; Dinara Shakiryanova
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Nature protocols     Volume:  2     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2007  
Date Detail:
Created Date:  2007-06-04     Completed Date:  2008-01-02     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1117-25     Citation Subset:  IM    
Affiliation:
Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA. elevitan@pitt.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Diagnostic Imaging / methods*
Drosophila / physiology*
Fluorescence Recovery After Photobleaching
Fourier Analysis
Green Fluorescent Proteins
Larva / physiology
Microscopy, Fluorescence
Neuromuscular Junction / physiology*
Neuropeptides / secretion*
Secretory Vesicles / secretion,  ultrastructure*
Synaptic Vesicles / secretion,  ultrastructure*
Grant Support
ID/Acronym/Agency:
NS32385/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Neuropeptides; 147336-22-9/Green Fluorescent Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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