Title | Visualizing and Analyzing Branching Microtubule Nucleation Using Meiotic Xenopus Egg Extracts and TIRF Microscopy. |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | King, M, Petry, S |
Journal | Methods Mol Biol |
Volume | 1413 |
Pagination | 77-85 |
Date Published | 2016 |
ISSN | 1940-6029 |
Keywords | Animals, Cell Division, Cell Nucleus, Genes, Reporter, Meiosis, Microscopy, Confocal, Microtubules, Mitosis, Oocytes, Recombinant Fusion Proteins, Spindle Apparatus, Staining and Labeling, Xenopus laevis |
Abstract | <p>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.</p> |
DOI | 10.1007/978-1-4939-3542-0_6 |
Alternate Journal | Methods Mol Biol |
PubMed ID | 27193844 |
PubMed Central ID | PMC5016078 |
Grant List | DP2 GM123493 / GM / NIGMS NIH HHS / United States R00 GM100013 / GM / NIGMS NIH HHS / United States T32 GM007388 / GM / NIGMS NIH HHS / United States |