Visualizing and analyzing branching microtubule nucleation using meiotic Xenopus egg extracts and TIRF microscopy

Matthew King, Sabine Petry

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Mitotic and meiotic spindles consist primarily of microtubules, which originate from centrosomes and within the vicinity of chromatin. Indirect evidence suggested that microtubules also originate throughout the spindle, but the high microtubule density within the spindle precludes the direct observation of this phenomenon. By using meiotic Xenopus laevis egg extract and employing total internal reflection (TIRF) microscopy, microtubule nucleation from preexisting microtubules could be demonstrated and analyzed. Branching microtubule nucleation is an ideal mechanism to assemble and maintain a mitotic spindle, because microtubule numbers are amplified while preserving their polarity. Here, we describe the assays that made these findings possible and the experiments that helped identify the key molecular players involved.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages77-85
Number of pages9
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1413
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Cell division
  • Cytoskeleton
  • Meiotic spindle
  • Microtubule
  • Microtubule nucleation
  • Mitotic spindle
  • TIRF microscopy
  • Xenopus laevis egg extract

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  • Cite this

    King, M., & Petry, S. (2016). Visualizing and analyzing branching microtubule nucleation using meiotic Xenopus egg extracts and TIRF microscopy. In Methods in Molecular Biology (pp. 77-85). (Methods in Molecular Biology; Vol. 1413). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3542-0_6