XB-ART-49689
J Vis Exp
September 7, 2014;
(91):
e52138.
Using plusTipTracker software to measure microtubule dynamics in Xenopus laevis growth cones.
Abstract
Microtubule (MT) plus-end-tracking proteins (+TIPs) localize to the growing plus-ends of MTs and regulate MT dynamics(1,2). One of the most well-known and widely-utilized +TIPs for analyzing MT dynamics is the End-Binding protein,
EB1, which binds all growing MT plus-ends, and thus, is a marker for MT polymerization(1). Many studies of
EB1 behavior within growth cones have used time-consuming and biased computer-assisted,
hand-tracking methods to analyze individual MTs(1-3). Our approach is to quantify global parameters of MT dynamics using the software package, plusTipTracker(4), following the acquisition of high-resolution, live images of tagged
EB1 in cultured embryonic growth cones(5). This software is a MATLAB-based, open-source, user-friendly package that combines automated detection, tracking, visualization, and analysis for movies of fluorescently-labeled +TIPs. Here, we present the protocol for using plusTipTracker for the analysis of fluorescently-labeled +TIP comets in cultured Xenopus laevis growth cones. However, this software can also be used to characterize MT dynamics in various cell types(6-8).
PubMed ID:
25225829
PMC ID:
PMC4189079
Article link:
J Vis Exp
Grant support:
[+]
R00 MH095768 NIMH NIH HHS
Species referenced:
Xenopus laevis
Genes referenced:
mapre1
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