Document Detail


Fluorescence characterization of the structural heterogeneity of polytene chromosomes.
MedLine Citation:
PMID:  19629653     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Studies on the physical nature of the structural heterogeneity of chromatin in their native states are few. The eukaryotic chromatin as observed by dye staining studies is of heterogeneous intensity when observed by fluorescent stains, where less and more bright regions apparently correspond to euchromatin and heterochromatin respectively. These are also associated with differential gene expression where it is believed that euchromatin is transcriptionally more active due to increased flexibility. Unfixed squashed preparations of polytene chromosomes of Drosophila were stained with a dsDNA specific dye PicoGreen and fluorescence lifetimes as well as fluorescence anisotropy decay kinetics were measured. Here we report a positive correlation between fluorescence lifetimes and fluorescence intensities, and show that less bright regions corresponding to euchromatin have shorter lifetimes, whereas more bright regions corresponding to heterochromatin have longer lifetimes. We interpret this as less bright regions being more dynamic, a conclusion also supported by fluorescence anisotropy decay kinetics. We infer that the comparatively higher flexibility associated with euchromatin can be directly measured by fluorescence lifetimes and fluorescence anisotropy decay kinetics.
Authors:
Sunil K Noothi; Mamata Kombrabail; Basuthkar J Rao; G Krishnamoorthy
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Publication Detail:
Type:  Journal Article     Date:  2009-07-24
Journal Detail:
Title:  Journal of fluorescence     Volume:  20     ISSN:  1573-4994     ISO Abbreviation:  J Fluoresc     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-02-10     Completed Date:  2010-05-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9201341     Medline TA:  J Fluoresc     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  37-41     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences, Tata Institute of Fundamental Research, Mumbai, 400005, India.
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MeSH Terms
Descriptor/Qualifier:
Animals
Chromatin / chemistry,  metabolism
Chromosomes / chemistry*,  metabolism
Drosophila melanogaster / cytology*
Fluorescence Polarization
Fluorescent Dyes / metabolism
Kinetics
Microscopy, Fluorescence
Organic Chemicals / metabolism
Chemical
Reg. No./Substance:
0/Chromatin; 0/Fluorescent Dyes; 0/Organic Chemicals; 0/PicoGreen

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