Document Detail

Trafficking of excitatory amino acid transporter 2-laden vesicles in cultured astrocytes: a comparison between approximate and exact determination of trajectory angles.
MedLine Citation:
PMID:  25408463     Owner:  NLM     Status:  Publisher    
A clear consensus concerning the mechanisms of intracellular secretory vesicle trafficking in astrocytes is lacking in the physiological literature. A good characterization of vesicle trafficking that may assist researchers in achieving that goal is the trajectory angle, defined as the angle between the trajectory of a vesicle and a line radial to the cell's nucleus. In this study, we provide a precise definition of the trajectory angle, describe and compare two methods for its calculation in terms of measureable trafficking parameters, and give recommendations for the appropriate use of each method. We investigated the trafficking of vesicles containing excitatory amino acid transporter 2 (EAAT2) fluorescently tagged with enhanced green fluorescent protein (EGFP) to quantify and validate the precision of each method. The motion of fluorescent puncta-taken to represent vesicles containing EAAT2-EGFP-was found to be typical of secretory vesicle trafficking. An exact method for calculating the trajectory angle of these puncta produced no error but required large computation time. An approximate method reduced the requisite computation time but produced an error depending on the inverse of the ratio of the punctum's initial distance from the nucleus centroid to its maximal displacement. Fitting this dependence to a power function allowed us to establish an exclusion distance from the centroid, beyond which the approximate method is less likely to produce an error above an acceptable 5 %. We recommend that the exact method be used to calculate the trajectory angle for puncta closer to the nucleus centroid than this exclusion distance.
Chapin E Cavender; Manoj K Gottipati; Vladimir Parpura
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-11-19
Journal Detail:
Title:  Amino acids     Volume:  -     ISSN:  1438-2199     ISO Abbreviation:  Amino Acids     Publication Date:  2014 Nov 
Date Detail:
Created Date:  2014-11-19     Completed Date:  -     Revised Date:  2014-11-20    
Medline Journal Info:
Nlm Unique ID:  9200312     Medline TA:  Amino Acids     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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