Document Detail


Analysis of the Trypanosoma brucei cell cycle by quantitative DAPI imaging.
MedLine Citation:
PMID:  18501977     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Trypanosoma brucei has two DNA compartments: the nucleus and the kinetoplast. DNA replication of these two compartments only partially coincides. Woodward and Gull [Woodward R, Gull K. Timing of nuclear and kinetoplast DNA replication and early morphological events in the cell cycle of Trypanosoma brucei. J Cell Sci 1990;95:49-57] comprehensively studied the relative timing of the replication and segregation of nuclear DNA (nDNA) and kinetoplast DNA (kDNA). Others have since assumed the consistency of morphological indicators of cell-cycle stage among strains and conditions. We report the use of quantitative DAPI imaging to determine the cell-cycle stage of individual procyclic cells. Using this approach, we found that kinetoplast elongation occurs mainly during nuclear S phase and not during G2, as previously assumed. We confirmed this finding by sorting cells by DNA content, followed by fluorescence microscopy. In addition, simultaneous quantitative imaging at two wavelengths can be used to determine the abundance of cell-cycle-regulated proteins during the cell cycle. We demonstrate this technique by co-staining for the non-acetylated state of lysine 4 of histone H4 (H4K4), which is enriched during nuclear S phase.
Authors:
T Nicolai Siegel; Doeke R Hekstra; George A M Cross
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-04-12
Journal Detail:
Title:  Molecular and biochemical parasitology     Volume:  160     ISSN:  0166-6851     ISO Abbreviation:  Mol. Biochem. Parasitol.     Publication Date:  2008 Aug 
Date Detail:
Created Date:  2008-06-23     Completed Date:  2008-09-11     Revised Date:  2010-12-03    
Medline Journal Info:
Nlm Unique ID:  8006324     Medline TA:  Mol Biochem Parasitol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  171-4     Citation Subset:  IM    
Affiliation:
Laboratory of Molecular Parasitology, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle*
Flow Cytometry
Image Processing, Computer-Assisted / methods*
Indoles / metabolism
Microscopy, Fluorescence
Staining and Labeling / methods*
Trypanosoma brucei brucei / physiology*
Grant Support
ID/Acronym/Agency:
AI 21729/AI/NIAID NIH HHS; R01 AI021729-23/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Indoles; 47165-04-8/DAPI
Comments/Corrections

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