The extraordinary robustness of mitotic spindle assembly to microtubule dynamics plasticity revealed by end-binding proteins and tubulin tagging
Abstract
Microtubules are dynamic, conserved cytoskeletal filaments that are essential for all eukaryotic cells. Microtubule dynamic properties are primarily characterized by measuring how fast they grow and shrink (growth and shrinkage rates) and how often they switch between assembly and disassembly (catastrophe and rescue frequencies). These four parameters measured for individual filaments can inform on the behavior of the entire microtubule network at the cell level. By comparing microtubule dynamics in Caenorhabditis elegans one-cell embryos using different genetically-encoded fluorescent probes, we observed an unexpected high variability in these parameters. Microtubule dynamics parameters were consistently higher in C. elegans strains expressing a fluorescently labelled microtubule end-binding protein than in strains relying on tubulin labelling, with microtubule growth rates differing by nearly a factor of two between the two conditions. This discrepancy was not limited to C. elegans , as we observed a similar effect in embryos of the tunicate Phallusia mammillata . Despite this, spindle size and assembly timing were only mildly affected. However, embryos expressing labelled end-binding protein exhibited higher frequency of mitotic defects and perturbed embryonic development upon exposure to various stresses such as elevated temperature or a compromised spindle assembly checkpoint. Thus, our work reveals both the remarkable robustness of mitotic spindle assembly in response to extreme microtubule dynamics plasticity, and the requirement for strict control of microtubule dynamics across successive early embryonic divisions. Our findings should also serve as a cautionary note when using tagged end-binding proteins to measure microtubule dynamics.
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BibTeX
@misc{robert2026extraordinary,
title = {{The extraordinary robustness of mitotic spindle assembly to microtubule dynamics plasticity revealed by end-binding proteins and tubulin tagging}},
author = {Morgane L.V. Robert and Aurélien Perrier and Janet Chenevert and Layla El-Mossadeq and Gilliane Maton and Alex Mcdougall and Anna Castro and Thierry Lorca and Julie Canman and Stefania Castagnetti and Julien Dumont and Benjamin Lacroix},
howpublished = {HAL},
year = {2026},
month = jul,
url = {https://hal.science/hal-05844355v1},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - UNPB
TI - The extraordinary robustness of mitotic spindle assembly to microtubule dynamics plasticity revealed by end-binding proteins and tubulin tagging
AU - Morgane L.V. Robert
AU - Aurélien Perrier
AU - Janet Chenevert
AU - Layla El-Mossadeq
AU - Gilliane Maton
AU - Alex Mcdougall
AU - Anna Castro
AU - Thierry Lorca
AU - Julie Canman
AU - Stefania Castagnetti
AU - Julien Dumont
AU - Benjamin Lacroix
PY - 2026
DA - 2026/07/30
PB - HAL
UR - https://hal.science/hal-05844355v1
AB - Microtubules are dynamic, conserved cytoskeletal filaments that are essential for all eukaryotic cells. Microtubule dynamic properties are primarily characterized by measuring how fast they grow and shrink (growth and shrinkage rates) and how often they switch between assembly and disassembly (catastrophe and rescue frequencies). These four parameters measured for individual filaments can inform on the behavior of the entire microtubule network at the cell level. By comparing microtubule dynamics in Caenorhabditis elegans one-cell embryos using different genetically-encoded fluorescent probes, we observed an unexpected high variability in these parameters. Microtubule dynamics parameters were consistently higher in C. elegans strains expressing a fluorescently labelled microtubule end-binding protein than in strains relying on tubulin labelling, with microtubule growth rates differing by nearly a factor of two between the two conditions. This discrepancy was not limited to C. elegans , as we observed a similar effect in embryos of the tunicate Phallusia mammillata . Despite this, spindle size and assembly timing were only mildly affected. However, embryos expressing labelled end-binding protein exhibited higher frequency of mitotic defects and perturbed embryonic development upon exposure to various stresses such as elevated temperature or a compromised spindle assembly checkpoint. Thus, our work reveals both the remarkable robustness of mitotic spindle assembly in response to extreme microtubule dynamics plasticity, and the requirement for strict control of microtubule dynamics across successive early embryonic divisions. Our findings should also serve as a cautionary note when using tagged end-binding proteins to measure microtubule dynamics.
ER -