Analysis of microtubule dynamic instability using a plus-end growth marker.
Regulation of microtubule dynamics is essential for many cell biological processes and is likely to be variable between different subcellular regions. We describe a computational approach to analyze microtubule dynamics by detecting growing microtubule plus ends. Our algorithm tracked all EB1-EGFP comets visible in an image time-lapse sequence allowing the detection of spatial patterns of microtubule dynamics. We introduce spatiotemporal clustering of EB1-EGFP growth tracks to infer microtubule behaviors during phases of pause and shortening. We validated the algorithm by comparing the results to data for manually tracked, homogeneously labeled microtubules and by analyzing the effects of well-characterized inhibitors of microtubule polymerization dynamics. We used our method to analyze spatial variations of intracellular microtubule dynamics in migrating epithelial cells.
Matov
Alexandre
A
Department of Cell Biology, The Scripps Research Institute, La Jolla, California, USA.
Applegate
Kathryn
K
Kumar
Praveen
P
Thoma
Claudio
C
Krek
Wilhelm
W
Danuser
Gaudenz
G
Wittmann
Torsten
T
D000107:Acetylation; D000465:Algorithms; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D019295:Computational Biology; D003198:Computer Simulation; D004847:Epithelial Cells / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000032:analysis* / Q000378:metabolism; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism*; D015203:Reproducibility of Results; D012984:Software; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism
D015415:Biomarkers; C405804:EB1 microtubule binding proteins; D008869:Microtubule-Associated Proteins; D014404:Tubulin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins