Visualizing Microtubule Networks During Drosophila Oogenesis Using Fixed and Live Imaging
Autor: | Kevin Legent, Antoine Guichet, Nicolas Tissot |
---|---|
Rok vydání: | 2015 |
Předmět: |
0303 health sciences
Polarity (physics) Biology Oocyte Cell biology 03 medical and health sciences 0302 clinical medicine medicine.anatomical_structure Tubulin Live cell imaging Cytoplasm Microtubule Cell polarity medicine biology.protein Cytoskeleton 030217 neurology & neurosurgery 030304 developmental biology |
Zdroj: | Methods in Molecular Biology ISBN: 9781493928507 |
Popis: | The microtubule cytoskeleton is a plastic network of polarized cables. These polymers of tubulin provide orientated routes for the dynamic transport of cytoplasmic molecules and organelles, through which cell polarity is established and maintained. The role of microtubule-mediated transport in the asymmetric localization of axis polarity determinants, in the Drosophila oocyte, has been the subject of extensive studies in the past years. However, imaging the distribution of microtubule fibers in a large cell, where vitellogenesis ensures the uptake of a thick and hazy yolk, presents a series of technical challenges. This chapter briefly reviews some of these aspects and describes two methods designed to circumvent these difficulties. We provide a detailed protocol for the visualization by immunohistochemistry of the three-dimensional organization of tubulin cables in the oocyte. Additionally, we detail the stepwise procedure for the live imaging of microtubule dynamics and network remodeling, using fluorescently labeled microtubule-associated proteins. |
Databáze: | OpenAIRE |
Externí odkaz: |