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Measuring nanometer scale gradients in spindle microtubule dynamics using model convolution microscopy.

Chad G Pearson1, Melissa K Gardner, Leocadia V Paliulis

  • 1Department of Molecular, Cellular, and Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309-0347, USA.

Molecular Biology of the Cell
|June 30, 2006
PubMed
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Researchers visualized tubulin dynamics in yeast mitotic spindles using advanced imaging. They found that kinetochore-attached microtubule (kMT) plus-end dynamics are crucial for accurate chromosome segregation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biophysics

Background:

  • Computational models predict spatial gradients in tubulin turnover within the budding yeast mitotic spindle.
  • Kinetochore-attached microtubules (kMTs) exhibit polymerization and depolymerization dynamics influencing this gradient.
  • Visualizing these dynamics is challenging due to the short length of kMTs in yeast, often below microscope resolution limits.

Purpose of the Study:

  • To overcome visualization limitations and measure the spatial gradient of tubulin dynamics in yeast mitotic spindles.
  • To investigate the relationship between microtubule dynamics, kinetochore tension, and chromosome segregation accuracy.

Main Methods:

  • Combined digital imaging of fluorescence redistribution after photobleaching (FRAP) with model convolution methods.

Related Experiment Videos

  • Compared nanometer-scale computer simulations with microscopic data.
  • Measured microtubule dynamics at approximately 65-nm spatial intervals within yeast spindles.

    • Main Results:

    • A spatial gradient in tubulin turnover was measured, with highest turnover near kinetochores and lowest near spindle poles.
    • A beta-tubulin mutant with reduced plus-end dynamics showed preserved tubulin turnover gradients over longer timescales.
    • This mutant exhibited a 14% increase in average kMT length and decreased kinetochore tension.

    Conclusions:

    • Robust kMT plus-end dynamics are essential for accurate chromosome segregation.
    • Altered microtubule dynamics in the beta-tubulin mutant led to an increased frequency of chromosome loss.
    • The study links microtubule dynamics directly to the fidelity of the cell division process.