Visualizing and Analyzing Branching Microtubule Nucleation Using Meiotic Xenopus Egg Extracts and TIRF Microscopy.

TitleVisualizing and Analyzing Branching Microtubule Nucleation Using Meiotic Xenopus Egg Extracts and TIRF Microscopy.
Publication TypeJournal Article
Year of Publication2016
AuthorsKing, M, Petry, S
JournalMethods Mol Biol
Volume1413
Pagination77-85
Date Published2016
ISSN1940-6029
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.

DOI10.1007/978-1-4939-3542-0_6
Alternate JournalMethods Mol. Biol.
PubMed ID27193844
PubMed Central IDPMC5016078
Grant ListR00 GM100013 / GM / NIGMS NIH HHS / United States