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Imaging septum formation by fluorescence microscopy

AuthorsRibas, Juan Carlos; Cortés, Juan Carlos G.
Glucan synthase
Calcofluor white
Cell wall
Issue Date2016
PublisherSpringer Nature
CitationYeast Cytokinesis. Methods and Protocols: 73-86 (2016)
SeriesMethods in Molecular Biology 1369
AbstractFungal cleavage furrow formation during cytokinesis relays in the coordinated contraction of an actomyosinbased ring and the centripetal synthesis of both new plasma membrane and a special wall structure named division septum. Through transmission electron microscopy, the septum exhibits a three-layered structure with a central primary septum, fl anked at both sides by the secondary septum. In contrast to the chitinous primary septum present in most of fungi, the fi ssion yeast Schizosaccharomyces pombe does not contain chitin, instead it divides through the formation of a linear β(1,3)glucan-rich primary septum, which has been shown to be specifi cally stained by the fl uorochrome Calcofl uor white. Recent fi ndings in S. pombe have revealed the importance of septum synthesis for the steady contraction of the ring during cytokinesis. Therefore, to study the molecular mechanisms that connect the extracellular septum wall with the other components of the cytokinetic machinery located in the plasma membrane and cytoplasm, new experimental approaches are needed. Here we describe the methods developed to image the septum structure by fl uorescence microscopy, with a special focus in the analysis of septum progression by the use of time-lapse microscopy.
Identifiersisbn: 978-1-4939-3144-6
Appears in Collections:(IBFG) Libros y partes de libros
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