Analysis of microtubule dynamic instability using a plus-end growth marker

Alexandre Matov; Kathryn Applegate; Praveen Kumar; Claudio Thoma; Wilhelm Krek; Gaudenz Danuser; Torsten Wittmann

doi:10.1038/nmeth.1493

Analysis of microtubule dynamic instability using a plus-end growth marker

Alexandre Matov, Kathryn Applegate, Praveen Kumar, Claudio Thoma, Wilhelm Krek, Gaudenz Danuser, Torsten Wittmann

Research output: Contribution to journal › Article › peer-review

182 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	761-768
Number of pages	8
Journal	Nature methods
Volume	7
Issue number	9
DOIs	https://doi.org/10.1038/nmeth.1493
State	Published - Sep 2010

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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title = "Analysis of microtubule dynamic instability using a plus-end growth marker",

abstract = "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.",

author = "Alexandre Matov and Kathryn Applegate and Praveen Kumar and Claudio Thoma and Wilhelm Krek and Gaudenz Danuser and Torsten Wittmann",

note = "Funding Information: This work was supported by US National Institutes of Health grants U01 GM067230 to G.D. and R01 GM079139 to T.W. This research was in part conducted in a facility constructed with support from the Research Facilities Improvement Program grant C06 RR16490 from the National Center for Research Resources of the National Institutes of Health. We thank A. Wheeler (Imperial College London) for the EB1-EGFP–expressing HaCaT cell line.",

year = "2010",

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doi = "10.1038/nmeth.1493",

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T1 - Analysis of microtubule dynamic instability using a plus-end growth marker

AU - Matov, Alexandre

AU - Applegate, Kathryn

AU - Kumar, Praveen

AU - Thoma, Claudio

AU - Krek, Wilhelm

AU - Danuser, Gaudenz

AU - Wittmann, Torsten

N1 - Funding Information: This work was supported by US National Institutes of Health grants U01 GM067230 to G.D. and R01 GM079139 to T.W. This research was in part conducted in a facility constructed with support from the Research Facilities Improvement Program grant C06 RR16490 from the National Center for Research Resources of the National Institutes of Health. We thank A. Wheeler (Imperial College London) for the EB1-EGFP–expressing HaCaT cell line.

PY - 2010/9

Y1 - 2010/9

N2 - 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.

AB - 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.

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Analysis of microtubule dynamic instability using a plus-end growth marker

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