title: 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).
authors.affiliation: Department of Biology, Boston College.
authors.forename: Alina
authors.initials: A
authors.lastname: Stout
authors.affiliation: Department of Biology, Boston College.
authors.forename: Salvatore
authors.initials: S
authors.lastname: D'Amico
authors.affiliation: Department of Biology, Boston College.
authors.forename: Tiffany
authors.initials: T
authors.lastname: Enzenbacher
authors.affiliation: Department of Biology, Boston College.
authors.forename: Patrick
authors.initials: P
authors.lastname: Ebbert
authors.affiliation: Department of Biology, Boston College; laura.lowery@bc.edu.
authors.forename: Laura Anne
authors.initials: LA
authors.lastname: Lowery
mesh_terms: D000818:Animals; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D020439:Growth Cones / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D012984:Software; D014982:Xenopus laevis
chemical_list: C405804:EB1 microtubule binding proteins; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins