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Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells

(2006) JOURNAL OF CELL SCIENCE. 119(15). p.3193-3205
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Abstract
The spindle occupies a central position in cell division as it builds up the chromosome-separating machine. Here we analysed the dynamics of spindle formation in acentrosomal plant cells by visualizing microtubules labelled with GFP-EB1, GFP-MAP4 and GFP-alpha-tubulin and chromosomes marked by the vital dye SYTO82. During prophase, few microtubules penetrate the nuclear area, followed by nuclear envelope disintegration. During prometaphase, microtubules invading the nuclear space develop a spindle axis from few bipolar microtubule bundles, which is followed by spindle assembly. Using a novel quantitative kymograph analysis based on Fourier transformation, we measured the microtubule growth trajectories of the entire dynamic metaphase spindle. Microtubules initiating from spindle poles either pass through the metaphase plate to form interpolar microtubule bundles or grow until they reach chromosomes. We also noticed a minor fraction of microtubules growing away from the chromosomes. Microtubules grow at 10 mu m/minute both at the spindle equator and at the spindle poles. Photobleached marks created on metaphase and anaphase spindles revealed a poleward tubulin flux. During anaphase, the velocity of tubulin flux (2 mu m/minute) equals the speed of chromatid-separation. With these findings we identified spatially coordinated microtubule growth dynamics and microtubule flux-based chromosome-separation as important facets of plant spindle operation.
Keywords
GFP, spindle, microtubule, flux, EB1, NUCLEAR-ENVELOPE BREAKDOWN, EGG EXTRACT SPINDLES, MITOTIC SPINDLE, PLUS-END, HAEMANTHUS ENDOSPERM, SELF-ORGANIZATION, MEIOTIC SPINDLES, POLEWARD FLUX, GAMMA-TUBULIN, ANAPHASE-A

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Chicago
Dhonukshe, Pankaj, Norbert Vischer, and Theodorus WJ Gadella Jr. 2006. “Contribution of Microtubule Growth Polarity and Flux to Spindle Assembly and Functioning in Plant Cells.” Journal of Cell Science 119 (15): 3193–3205.
APA
Dhonukshe, P., Vischer, N., & Gadella Jr, T. W. (2006). Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells. JOURNAL OF CELL SCIENCE, 119(15), 3193–3205.
Vancouver
1.
Dhonukshe P, Vischer N, Gadella Jr TW. Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells. JOURNAL OF CELL SCIENCE. 2006;119(15):3193–205.
MLA
Dhonukshe, Pankaj, Norbert Vischer, and Theodorus WJ Gadella Jr. “Contribution of Microtubule Growth Polarity and Flux to Spindle Assembly and Functioning in Plant Cells.” JOURNAL OF CELL SCIENCE 119.15 (2006): 3193–3205. Print.
@article{3202726,
  abstract     = {The spindle occupies a central position in cell division as it builds up the chromosome-separating machine. Here we analysed the dynamics of spindle formation in acentrosomal plant cells by visualizing microtubules labelled with GFP-EB1, GFP-MAP4 and GFP-alpha-tubulin and chromosomes marked by the vital dye SYTO82. During prophase, few microtubules penetrate the nuclear area, followed by nuclear envelope disintegration. During prometaphase, microtubules invading the nuclear space develop a spindle axis from few bipolar microtubule bundles, which is followed by spindle assembly. Using a novel quantitative kymograph analysis based on Fourier transformation, we measured the microtubule growth trajectories of the entire dynamic metaphase spindle. Microtubules initiating from spindle poles either pass through the metaphase plate to form interpolar microtubule bundles or grow until they reach chromosomes. We also noticed a minor fraction of microtubules growing away from the chromosomes. Microtubules grow at 10 mu m/minute both at the spindle equator and at the spindle poles. Photobleached marks created on metaphase and anaphase spindles revealed a poleward tubulin flux. During anaphase, the velocity of tubulin flux (2 mu m/minute) equals the speed of chromatid-separation. With these findings we identified spatially coordinated microtubule growth dynamics and microtubule flux-based chromosome-separation as important facets of plant spindle operation.},
  author       = {Dhonukshe, Pankaj and Vischer, Norbert and Gadella Jr, Theodorus WJ},
  issn         = {0021-9533},
  journal      = {JOURNAL OF CELL SCIENCE},
  keyword      = {GFP,spindle,microtubule,flux,EB1,NUCLEAR-ENVELOPE BREAKDOWN,EGG EXTRACT SPINDLES,MITOTIC SPINDLE,PLUS-END,HAEMANTHUS ENDOSPERM,SELF-ORGANIZATION,MEIOTIC SPINDLES,POLEWARD FLUX,GAMMA-TUBULIN,ANAPHASE-A},
  language     = {eng},
  number       = {15},
  pages        = {3193--3205},
  title        = {Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells},
  url          = {http://dx.doi.org/10.1242/jcs.03048},
  volume       = {119},
  year         = {2006},
}

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