Eukaryotic Cell doi:10.1128/EC.00415-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.
Cell cycle synchrony in Giardia intestinalis cultures using nocodazole and aphidicolin
Marianne Poxleitner,
Scott Dawson,
and
W. Zacheus Cande*
Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA 94720; Section of Microbiology, 255 Briggs Hall, One Shields Avenue, University of California at Davis, Davis, CA 95616
* To whom correspondence should be addressed. Email:
zcande{at}berkeley.edu.
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Abstract |
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Giardia intestinalis is a ubiquitous intestinal protozoan parasite and has been proposed to represent the earliest diverging lineage of extant eukaryotes. Despite its importance as a model organism, research on Giardia has been hampered by an inability to achieve cell cycle synchrony for in vitro cultures. This research details successful methods for attaining cell cycle synchrony in Giardia cultures. Research presented here demonstrates reversible cell cycle arrest in G1/S and G2/M with aphidicolin and nocodazole, respectively. Following synchronization, cells are able to recover completely from drug treatment and remain viable and maintain synchronous growth for six hours. These techniques are used to synchronize Giardia cultures to increase the percentage of mitotic spindles in culture. This method of synchronization will enhance our ability to study cell cycle dependant processes in Giardia intestinalis.
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