Imaging and Quantifying the Dynamics of γ-Tubulin at Microtubule Minus Ends in Mitotic Spindles

Methods Mol Biol. 2016:1413:63-75. doi: 10.1007/978-1-4939-3542-0_5.

Abstract

Understanding the organization of complex microtubule arrays such as the mitotic spindle requires information about the position and dynamics of microtubule plus and minus ends. Whereas plus end dynamics have been widely studied using markers such as EB1-GFP, much less is known about the dynamic properties of minus ends, in part because a suitable marker has only recently become available. Here we describe the use of photoactivatable γ-tubulin-paGFP to image and quantify the dynamics of microtubule minus ends in mitotic spindles.

Keywords: Microtubule; Minus end dynamics; Photoactivatable GFP; Spindle; γ-Tubulin.

MeSH terms

  • Cell Line
  • Gene Expression
  • Genes, Reporter
  • Humans
  • Light
  • Metaphase
  • Microscopy, Fluorescence
  • Microtubules / metabolism*
  • Molecular Imaging* / methods
  • Quinazolinones / pharmacology
  • RNA Interference
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Spindle Apparatus / metabolism*
  • Transcriptional Activation / radiation effects
  • Tubulin / metabolism*

Substances

  • Quinazolinones
  • Recombinant Fusion Proteins
  • Tubulin
  • ciliobrevin D