On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells

Microtubules are dynamic structures undergoing rapid growth and shrinkage in living cells and in vitro. The growth of microtubules in vitro was analyzed with subpixel precision (Maurer et al., Current Biology, 2014, 24 (4), 372–384); however, to what extent these results could be applied for microtu...

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Autores principales:	Arshat Urazbaev, Anara Serikbaeva, Anna Tvorogova, Azamat Dusenbayev, Sholpan Kauanova, Ivan Vorobjev
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	microtubules dynamic instability EB proteins fluorescent microscopy cell culture live cell imaging Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/1f4d48a579ea41f4b40530488c920c1f
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spelling	oai:doaj.org-article:1f4d48a579ea41f4b40530488c920c1f2021-11-08T05:18:56ZOn the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells2296-889X10.3389/fmolb.2021.745089https://doaj.org/article/1f4d48a579ea41f4b40530488c920c1f2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmolb.2021.745089/fullhttps://doaj.org/toc/2296-889XMicrotubules are dynamic structures undergoing rapid growth and shrinkage in living cells and in vitro. The growth of microtubules in vitro was analyzed with subpixel precision (Maurer et al., Current Biology, 2014, 24 (4), 372–384); however, to what extent these results could be applied for microtubules growing in vivo remains largely unknown. Particularly, the question is whether microtubule growth velocity in cells could be sufficiently approximated by a Gaussian distribution or its variability requires a more sophisticated description? Addressing this question, we used time-lapse microscopy and mathematical modeling, and we analyzed EB-3 comets forming on microtubules of cultured cells with subpixel precision. Parameters of comets (shape, form, and velocity) were used as topological characteristics of 3D voxel objects. Using regression analysis, we determined the real positions of the microtubule tips in time-lapse sequences. By exponential decay fitting of the restored comet intensity profile, we found that in vivo EB-3 rapidly exchanges on growing microtubule ends with a decoration time ∼ 2 s. We next developed the model showing that the best correlation between comet length and microtubule end growth velocity is at time intervals close to the decoration time. In the cells, EB comet length positively correlates with microtubule growth velocity in preceding time intervals, while demonstrating no correlation in subsequent time intervals. Correlation between comet length and instantaneous growth velocity of microtubules remains under nocodazole treatment when mean values of both parameters decrease. Our data show that the growth of microtubules in living cells is well-approximated by a constant velocity with large stochastic fluctuations.Arshat UrazbaevAnara SerikbaevaAnara SerikbaevaAnna TvorogovaAzamat DusenbayevSholpan KauanovaSholpan KauanovaIvan VorobjevIvan VorobjevIvan VorobjevFrontiers Media S.A.articlemicrotubulesdynamic instabilityEB proteinsfluorescent microscopycell culturelive cell imagingBiology (General)QH301-705.5ENFrontiers in Molecular Biosciences, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	microtubules dynamic instability EB proteins fluorescent microscopy cell culture live cell imaging Biology (General) QH301-705.5
spellingShingle	microtubules dynamic instability EB proteins fluorescent microscopy cell culture live cell imaging Biology (General) QH301-705.5 Arshat Urazbaev Anara Serikbaeva Anara Serikbaeva Anna Tvorogova Azamat Dusenbayev Sholpan Kauanova Sholpan Kauanova Ivan Vorobjev Ivan Vorobjev Ivan Vorobjev On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
description	Microtubules are dynamic structures undergoing rapid growth and shrinkage in living cells and in vitro. The growth of microtubules in vitro was analyzed with subpixel precision (Maurer et al., Current Biology, 2014, 24 (4), 372–384); however, to what extent these results could be applied for microtubules growing in vivo remains largely unknown. Particularly, the question is whether microtubule growth velocity in cells could be sufficiently approximated by a Gaussian distribution or its variability requires a more sophisticated description? Addressing this question, we used time-lapse microscopy and mathematical modeling, and we analyzed EB-3 comets forming on microtubules of cultured cells with subpixel precision. Parameters of comets (shape, form, and velocity) were used as topological characteristics of 3D voxel objects. Using regression analysis, we determined the real positions of the microtubule tips in time-lapse sequences. By exponential decay fitting of the restored comet intensity profile, we found that in vivo EB-3 rapidly exchanges on growing microtubule ends with a decoration time ∼ 2 s. We next developed the model showing that the best correlation between comet length and microtubule end growth velocity is at time intervals close to the decoration time. In the cells, EB comet length positively correlates with microtubule growth velocity in preceding time intervals, while demonstrating no correlation in subsequent time intervals. Correlation between comet length and instantaneous growth velocity of microtubules remains under nocodazole treatment when mean values of both parameters decrease. Our data show that the growth of microtubules in living cells is well-approximated by a constant velocity with large stochastic fluctuations.
format	article
author	Arshat Urazbaev Anara Serikbaeva Anara Serikbaeva Anna Tvorogova Azamat Dusenbayev Sholpan Kauanova Sholpan Kauanova Ivan Vorobjev Ivan Vorobjev Ivan Vorobjev
author_facet	Arshat Urazbaev Anara Serikbaeva Anara Serikbaeva Anna Tvorogova Azamat Dusenbayev Sholpan Kauanova Sholpan Kauanova Ivan Vorobjev Ivan Vorobjev Ivan Vorobjev
author_sort	Arshat Urazbaev
title	On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
title_short	On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
title_full	On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
title_fullStr	On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
title_full_unstemmed	On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells
title_sort	on the relationship between eb-3 profiles and microtubules growth in cultured cells
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/1f4d48a579ea41f4b40530488c920c1f
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On the Relationship Between EB-3 Profiles and Microtubules Growth in Cultured Cells

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